Advertisement

TNF-α G-308A genetic variants, serum CRP-hs concentration and DNA damage in obese women

  • 753 Accesses

Abstract

Obesity is associated with inflammation, which can disturb genome stability. Tumor necrosis factor (TNF-α) polymorphism was found to affect TNF-α protein production and inflammation. Therefore, the present study illustrates the relationship between TNF-α polymorphism, the degree of inflammation assessed by serum high sensitivity C-reactive protein concentration (CRP-hs) and basal DNA damage in patients with obesity (BMI 30–34.9 kg/m2) and control subjects with proper body mass (BMI < 25 kg/m2). A total of 115 participants (75 obese premenopausal women; and 40 age-, and gender-matched controls) were included. Biochemical parameters (serum concentrations of total-cholesterol, HDL-cholesterol, LDL- cholesterol, triglycerides, glucose, apolipoprotein AI, CRP-hs) and endogenous DNA damage (determined by comet assay) were measured. TNF-α G-308A polymorphism (rs1800629) was analyzed by PCR-RFLP (PCR-restriction fragments length polymorphism). An effect of TNF-α genotype on serum CRP-hs concentration was noted (p = 0.031). In general, carriers of the rare A allele of the TNF-α G-308A polymorphism had significantly lower endogenous DNA damage and serum CRP-hs concentrations than GG homozygotes, however, the protective effect of the A allele was especially visible in non-obese women. Serum CRP-hs concentrations and levels of DNA damage (% DNA in tail) were significantly higher in obese than in controls (p = 0.001 and p < 0.0001, respectively). The adjusted multiple linear regression analyses revealed a significant, independent impact of obesity on DNA damage (p = 0.00000) and no effect of other covariates i.e. age, TNF-α genotype and serum CRP-hs concentration. Our study showed that obesity has a significant impact on the levels of endogenous DNA damage. Obesity abolished the protective effect of A allele of the TNF-α G-308A polymorphism on DNA damage and on inflammation development observed in non-obese A allele carriers.

Introduction

Smoking, improper diet and environmental toxins have been recognized as main exogenous sources of DNA damage [1]. However, besides exogenous factor-caused DNA breaks, endogenous DNA damage and failure of DNA repair can disturb cell metabolism and function [2]. In cells, production of reactive oxygen species (ROS) and inflammation have been recognized to cause DNA lesions [3]. DNA damage has been found to be involved in aging and development of common diseases including cancer, atherosclerosis, metabolic syndrome [4,5,6].

The Comet Assay is a sensitive and low-cost technique, which measures DNA damage in individual cells [7, 8]. In addition to DNA strand breaks (double strand breaks and single strand breaks), the modified Comet Assay serve to detect also oxidized bases, interstrand cross-links or misincorporated uracil [9]. This method is widely used to determine the level of DNA damage, both resulting from exposure to environmental mutagens, as well as arising in the course of many diseases [10,11,12].

Obesity is a worldwide problem with increasing prevalence, associated with co-morbidities such as type 2 diabetes mellitus and cardiovascular diseases, and increased cancer risk [13,14,15]. Obesity is characterized by the, adipocyte hypertrophy, elevated production of reactive oxygen species, cytokines, chronic inflammation, disturbances in insulin and glucose metabolism [16,17,18]. Association between the occurrence of DNA lesions and enhanced body weight has been also reported [19,20,21]. Both inflammation and metabolic disturbances can cause DNA damage [22,23,24]. Moreover, the relationship between chronic inflammation and genomic instability has been observed in about 25% of human cancers [25,26,27].

Tumor necrosis factor-alpha (TNF-α) is a multi-functional cytokine synthesized by adipocytes, preadipocytes, endothelial cells, smooth muscle cells, fibroblasts, leukocytes and macrophages [28,29,30,31]. It can participate in regulation of many cellular processes such as immune function, differentiation, proliferation, apoptosis and energy pathways [32, 33]. Variations in the TNF-α gene can affect TNF-α production and a significant effect of the polymorphism in the promoter region of the TNF-α gene at position -308 (rs1800629) was reported [34,35,36]. Presence of the variant allele has been shown to increase the rate of transcription and production of the TNF-α protein [37,38,39]. G-308A polymorphism in the TNF-α gene has been associated with the development of inflammation and risk of cardiovascular diseases [40,41,42,43,44]. A recent study revealed the predisposition of GG homozygotes to higher production of pro-inflammatory molecules resulting in their enhanced serum levels [45]. The G-308A polymorphism was also found to be associated with insulin sensitivity and increased production of leptin, suggesting an impact of TNF-α gene on obesity and obesity-related health complications [46]. Phillips et al. showed that patients carrying the GG genotype had elevated risk of metabolic syndrome compared with carriers of the minor A allele [47]. However, large cohort studies in Chinese, Caucasians and Afro-Americans did not show a significant correlation between G-308A polymorphism and insulin resistance or obesity [48,49,50,51]. Recently published systematic review and meta-analysis have indicated an association between TNF-α G-308A gene polymorphism and the risk of ischemic heart disease [28, 52].

TNF-α stimulates the production of C-reactive protein (CRP) and the development of inflammatory processes, and serum CRP concentration is commonly used as a marker of inflammation [53,54,55,56]. Furthermore, in vitro studies showed increased DNA damage as a result of TNF-α stimulated ROS production [57]. TNF-α together with IL-1β and IFN-γ induced DNA damage in human cholangiocarcinoma cell line [58]. DNA damage and enhanced ROS levels were related to TNF-α—mediated senescence in HUVEC (human umbilical vein endothelial cells) [59].

The aim of our study was to assess the impact of the G-308A TNF-α polymorphism on CRP-hs levels and genomic stability measured by basal DNA damage in obese women.

Materials and methods

Subjects

The study participants were premenopausal women (Polish Caucasians from the Warsaw region). Among 115 participants of the study, 75 were obese. Obesity was classified according to World Health Organization criteria [60] i.e., subjects with BMI ≥ 30 kg/m2 were considered obese. The obese women were consecutively recruited between December 2011 and June 2013 on the basis of clinical assessments from subjects who had been directed to the Outpatient Clinic at the National Food and Nutrition Institute in Warsaw due to obesity treatment. The gender- and age-matched control group (n = 40) of apparently healthy women with proper body mass (BMI not exceeded 25 kg/m2) was recruited from subjects directed for a routine general health screening. The study was conducted according to the guidelines laid down in the Declaration of Helsinki and the Local Ethics Committee at the National Food and Nutrition Institute approved all procedures involving human subjects. Written informed consent was obtained from all of the registered volunteers.

The recruited women were premenopausal, non- smoking (for at least 5 years), had no history of alcoholism, and had no signs or symptoms of renal and hepatic disorders, endocrine disorders (e.g. disease of the thyroid, parathyroid, Cushing’s syndrome, polycystic ovary syndrome), autoimmune diseases, and cancer. Women within the last 3 months before the study were not receiving medications known to influence plasma lipid levels and did not use hormonal therapy as well as did not report chronic use of dietary supplements and anti-inflammatory drugs. Exclusion factors were also menopause, pregnancy, and lactation.

Anthropometric measurements

All subjects underwent a comprehensive medical evaluation including medical history, physical examination and measurement of anthropometric parameters: body weight, body height, waist circumference, hip circumference according to standardized procedures routinely performed in the Outpatient Clinic at the National Food and Nutrition Institute (Warsaw, Poland). The body waist circumference was measured at the midpoint between the lower margin of the last rib cage and the top iliac crest by using a flexible inch tape. Measurements were taken in the morning, after an overnight fasting, at the same day, or the day before blood samplings. Based on anthropometric measurements the BMI and WHR (waist-hip ratio) indexes were calculated.

Blood analysis

Blood was collected after night fasting from all subjects and serum parameters were analyzed on the same day. Total cholesterol, HDL-cholesterol, triglycerides, glucose, and insulin were measured using standard techniques in a certified laboratory for clinical chemistry at The National Food and Nutrition Institute. The LDL cholesterol levels were calculated using the Friedewald formula. Residue serum was aliquoted and frozen at − 20 °C until analysis. CRP-hs concentrations were obtained using commercially available ELISA (Immundiagnostik AG, Germany), according to the protocol provided by the manufacturer. The serum concentrations of apolipoprotein AI were measured using monoclonal antibodies against apolipoprotein AI (Pointe Scientific, USA) by the immunotubidymetric method.

Genotype analysis

Genomic DNA was extracted from peripheral white blood cells of whole-blood samples using DNA Mini Kit (A&A Biotechnology, Poland). Extracted DNA samples (100 ng) were amplified to obtain a fragment including the polymorphic region of TNF-α G-308A gene (rs 1,800,629), as previously described [61] by using the following primers: F5′-AATAGGTTTTGAGGGCCATG-3′ and R5′-GGGACACACAAGCATCAAGG-3′. Approximately 100 ng of DNA was amplified by thermal cycling using the DNA polymerase kit (BioLine, London, UK) in 25 µL of PCR mixture containing 2.5 mM MgCl2, 0.4 mM of each deoxyribonucleotide triphosphate (dNTP, New England Biolabs, USA), 1 U Taq, and 100 pM of each primer. Polymerase chain reaction conditions included an initial denaturation at 94 °C for 5 min followed by 35 cycles of 94 °C for 15 s, 55 °C for 30 min, and 72 °C for 15 s, with a final extension at 72 °C for 7 min. The amplified DNA samples containing a polymorphic site was digested with the restriction enzyme NcoI (New England Biolabs, USA) and products were run on agarose gel electrophoresis. Digestion of the 151 bp fragment carrying the G allele was giving 139 bp and 12 bp fragments, while the fragment with the A allele remained intact. About 20% of all samples were randomly selected for repeated genotyping for confirmation. Concordance between repeats was 100%.

Comet assay

DNA integrity was determined by the use of alkaline single-cell gel electrophoresis (comet assay), based on previous reports [62, 63]. Lymphocytes were obtained from 1 mL heparinized blood by centrifugation in a density gradient; then 50 µL of lymphocytes (1–3 × 105 cells/mL) was distributed with 50 µL of 2% low-melting-point agarose on a microscope slide precoated with 0.5% normal agarose. The slides were incubated for 1 h in a freshly prepared cold (4 °C) lysis solution (2.5 M NaCl, 100 mM EDTA-Na2, 10 mM Tris, pH 10.0–10.5) with 1% Triton X-100. Next, the slides were left in a horizontal gel electrophoresis tank with alkaline electrophoresis buffer (300 mM NaOH, 1 mM EDTA-Na2, pH > 13.0) for 40 min at 4 °C. Electrophoresis was performed under following conditions: 20 min, 35 V (1 V/cm), 300 mA. Slides were then washed with a neutralizing solution (0.4 M Tris, pH 7.5), and stained with DAPI (20 µg/ml).Nikon Eclipse 50i fluorescence microscope (×400 magnification) and Lucia Comet Assay software version 4.81 (Laboratory Imaging, Prague, Czech Republic) was used to analyze 100 comets on each slide. From each subject three blood samples were analyzed in duplicates. Of the data obtained, % DNA in the tail was chosen for further analysis as a DNA damage parameter. The chemicals were supplied by Sigma–Aldrich.

Statistical analysis

All statistical calculations were performed with the Statistica software (version 12.0). The distribution of variables was tested by Shapiro–Wilk test. Differences in continuous parameters were tested using Mann–Whitney U-test. Spearman correlation analyses were performed for the relationships among the variables. Non-continuous variables were tested with a Chi square test. Allele frequencies for TNF-α variants were calculated with the gene counting method. Hardy–Weinberg equilibrium (HWE) was determined by Pearson’s χ2 goodness-of-fit test. CRP-hs was dichotomized as ≥ 3 mg/L versus otherwise (< 3 mg/L) based on the well accepted cut-off point (of 3 mg/L) indicating elevated CRP-hs associated with an increased risk for CVD [64]. Regression analyses were performed using generalized linear models. Odds ratios (OR) with 95% confidence intervals (95% CI) were calculated using logistic regression. Results were expressed as means ± SD or percentages, and p < 0.05 was considered statistically significant.

Results

Characteristics of the studied subjects are summarized in Table 1. Obese and control subjects (non-obese) were in similar age. There were substantial differences in serum concentrations of total cholesterol, triglycerides, LDL-cholesterol, and blood pressure between studied groups (p < 0.05). While no differences in HDL-cholesterol, apolipoprotein AI and glucose concentrations were found. Serum C-reactive protein (CRP-hs) concentrations as well as mean level of DNA damage (% DNA in tail) were significantly higher in obese than in controls (Table 1).

Table 1 Clinical and biochemical characteristics of the study population

Among all studied subjects the distribution of TNF-α gene alleles was in Hardy–Weinberg equilibrium (χ2 value = 2.68, p = 0.100). The frequency of the rare A allele of the TNF-α gene G-308A polymorphism was 17.4%. 71% of the studied women carried the GG genotype (wildtype), 24%—the GA genotype and 5%—the AA genotype. As reported in Table 2, no statistically significant difference in frequency of the three genotypes of G-308A TNF-α polymorphism among obese and non-obese was found. However, the prevalence of obesity was higher among subjects with GG genotype than among A allele carriers (71% and 50% respectively, p = 0.026). Due to the low frequency of the AA genotype statistical analyses were performed for A allele carriers (GA and AA genotypes pooled) and GG homozygotes.

Table 2 The distribution of genotypes and alleles of TNF-α G-308A polymorphism in obese and non-obese subjects

Study participants’ characteristics according to the G-308A TNF-α polymorphism are presented in Table 3. In GG homozygotes higher diastolic blood pressure was observed. No statistically significant difference in mean values of BMI and WHR between analyzed groups was recognized. Carriers of the GG genotype had higher levels of DNA damage as well as higher CRP-hs serum concentrations compared to the A allele carriers (3.55 ± 1.70% vs. 2.80 ± 1.29%; p = 0.025 and 4.06 ± 2.07 mg/L vs. 2.92 ± 2.15 mg/L, respectively, p = 0.0001).

Table 3 Clinical and biochemical characteristics of the study population according to TNF-α G-308A gene polymorphism

In studied group DNA damage was significantly correlated with BMI, WHR, systolic and diastolic blood pressure as well as total cholesterol, LDL cholesterol, and serum CRP-hs concentrations (Table 4). The observed association between DNA damage and BMI was not affected by the TNF-α genotype. However, an impact of the TNF-α genotype on the associations between DNA damage and WHR and serum CRP-hs was observed. Only among A allele carriers a strong, positive correlation between DNA damage and WHR was observed (R = 0.784, p = 0.00004). Also in A allele carriers but not in GG homozygotes, a positive correlation between DNA damage and serum CRP-hs concentration was recognized (Table 4, R = 0.578, p = 0.0003).

Table 4 Spearman correlations between DNA damage (% DNA in tail) and biochemical and anthropometric parameters

When obese and non-obese carriers of studied genotypes were analyzed separately, obesity not TNF-α genotype was found to affect DNA damage as higher levels of DNA damage occurred in obese compared to non-obese. Obese women carrying GG genotype had only slightly more DNA damage (% DNA in tail) than obese A allele carriers (4.37 ± 1.27% and 3.84 ± 0.97%, respectively; p = 0.084). Also among non-obese women non-significant allele effect on DNA damage was observed (1.49 ± 0.43% in GG and 1.75 ± 0.41% in A allele carriers; p = 0.057). The G-308A TNF-α polymorphism was found to affect serum CRP-hs concentrations only in non-obese women. Mean serum CRP-hs concentration was significantly higher in non-obese women with GG genotype than in non-obese A allele carriers (p = 0.0001), while among obese participants similar CRP-hs levels were observed (Fig. 1).

Fig. 1
figure1

CRP-hs levels among studied groups. A significant difference in CRP-hs levels between GG homozygotes and A allele carriers of TNF-α G-308A polymorphism was seen among controls (non-obese) but not among obese

Taking into account the degree of inflammation, higher, but not significant incidence of elevated serum CRP-hs concentrations (≥ 3 mg/L) was found among obese subjects: (OR 1.97 95% CI 0.88–4.41, p = 0.095). Additionally, logistic regression analysis identified GG genotype as a risk factor for elevated CRP-hs (≥ 3 mg/L) only in non-obese women (Table 5). The odds of CRP-hs ≥ 3 mg/L in non-obese women with GG genotype was 50 times greater than in non-obese women with A allele (p = 0.0001, OR 50.00, 95% CI 6.95–359.75). Multiple linear regression analyses revealed also a significant interaction effect of TNF-α genotype and elevated CRP-hs (≥ 3 mg/L) on the levels of DNA damage (F = 4.75, p = 0.031) in all studied women. However, when all covariates elevated CRP-hs, TNF-α genotype, obesity and age were included into the statistical analyses, the impact of obesity (BMI ≥ 30 kg/m2) on the level of DNA damage was independent of other covariates (F = 69.41, p = 0.0000) and no significant interaction effect was observed.

Table 5 Relationship between obesity, TNF-α polymorphism and elevated CRP-hs concertation in serum (≥ 3 mg/L)

Discussion

Obesity, that is a result of an imbalance between energy intake and expenditure, has reached epidemic proportions with increasing prevalence worldwide. Adipose tissue participates in the production of inflammatory mediators, and in adipose tissue from obese enhanced TNF-α production was observed [51, 52, 65,66,67].

The G-308A polymorphism in the promoter region of the TNF-α gene was found to affect TNF-α protein expression and ischemic heart disease risk in, both, Caucasians and Asians [68, 69]. The human TNF-α protein is coded by the gene located near major histocompatibility complex (MHC) between the class I HLA-B and the class II HLA-DR loci [70, 71]. Therefore, the SNPs in the TNF-α promoter may be related to HLA haplotypes and autoimmune diseases such as systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA) [72,73,74].

Low-grade chronic inflammation is a characteristic feature of obesity, and plays an important role in the pathogenesis of obesity-associated comorbidities [16]. Inflammation is linked with enhanced generation of reactive oxygen species (ROS), which can damage cellular biomolecules, including DNA, leading to disturbances in cell signaling and cell cycle control, genetic mutations, and promotion of inflammation [75].

In vitro studies recognized that pro-inflammatory cytokines provoke DNA damage, cell senescence and growth arrest [59, 76, 77]. In IFNγ/TNFα-induced genotoxicity, NADPH oxidases (Nox 1 and 4) and TGFβ/SMAD pathways are involved in enhanced ROS production [76]. ROS formation and increased level of DNA lesions were observed as a result of high CRP-hs in the culture of HUVECs [78]. Oxidative stress, chronic inflammation and DNA damage have been recognized as important factors leading to the development of carcinogenesis, atherosclerosis and cardiovascular diseases. Obesity is associated with elevated risk of all these diseases [79,80,81,82,83]. Therefore, not only obesity-associated inflammation but also obesity-associated DNA damage may play a significant role in the development of both cardiovascular diseases and cancer in obese [26, 84]. In cancerogenesis, enhanced mutation rate was found to be linked to a high amount of DNA lesions [85, 86].

The G-308A polymorphism in TNF-α gene was reported in relation to TNF-α protein production and development of inflammation as well as it was suggested to play an important role in the development and progression of cancer [87,88,89]. Therefore, this polymorphism may affect both development of inflammation and formation of DNA lesions. Thus, we hypothesized that in obese amount of endogenous DNA lesions may be linked to the degree of inflammation and TNF-α gene polymorphism, and in the present study we assessed the association between G-308A TNF-α gene variants, serum concentrations of CRP-hs and DNA damage in obese.

The present study was conducted in Caucasian women from the central region of Poland and among our study participants we found the low frequency of the AA genotype (5%) as well as A allele (17.4%) of the G-308A polymorphism in TNF-α gene. This is consistent with the results of other studies reporting that the G to A change in TNF-α gene is rather rare [90]. In a study of 120 Caucasian Italian women no AA homozygotes were recognized and the frequency of A allele was 27.4% [90]. The frequency of AA genotype was about 4% among Han Chinese [91], 2% among people with obesity from Spain [92], and 0.6% in the Brazilian individuals [93]. In GG homozygotes higher plasma levels of TNF-α and CRP-hs than in AA homozygotes [94,95,96,97] as well as in GA heterozygotes of the G-308A polymorphism in the TNF-α gene were reported [90].

In the present study non-obese carriers of the A allele had significantly lower CRP-hs serum concentrations than GG homozygotes. The presence of the A allele appears to have a protective anti-inflammatory effect, which, however, disappears when obesity appears. We observed similar CRP-hs concentrations in both obese and non-obese GG homozygotes, while obese had higher levels of DNA damage. It indicates that in obese GG homozygotes other factors than inflammation, have a significant impact on cellular DNA damage. In A allele carriers DNA damage was positively correlated with serum CRP-hs concentration and in obese-A allele carriers similar levels of serum CRP-hs and DNA damage as in obese-GG homozygotes were observed. Our study participants, both obese and controls, can be classified as apparently healthy, thus we can hypothesize that obesity and low-grade inflammation characteristic for obesity can affect basal DNA damage observed in this study. We found significantly greater amount of DNA lesions in obese than in non-obese women as well as the associations between BMI and DNA damage. It is in agreement with our previous study [21]. However, the results of the presented study show that the impact of obesity and obesity-associated disturbances on DNA damage is strong and occurrence of obesity eliminates or significantly decreases the effect of the G-308A TNF-α variants on both inflammation, and levels of DNA damage. In obesity oxidative stress and inflammation are involved in the induction of DNA lesions and have an impact on the efficiency of the DNA repair mechanisms [24]. DNA damage in cells may be induced by pro-inflammatory cytokines, chemokines and molecules such as NO (nitric oxide), and ROS [98, 99]. In white adipose tissue amount of DNA damage was related to pro-inflammatory markers such as IL-6 and TNF-α [77]. As CRP-hs exerts ROS production in vitro [78] it can be hypothesized that the potential cause of DNA damage found in our study participants is oxidative stress related to enhanced inflammation (CRP-hs ≥ 3 mg/dl). Our results indicates that in non-obese, apparently healthy women GG homozygosity of the G-308A TNF-α polymorphism is associated with enhanced low grade inflammation assessed by serum CRP-hs concentrations, and occurrence of obesity does not affect significantly CRP-hs levels in GG homozygotes. The presence of A allele in non-obese women protects against inflammation but development of obesity abolished this allele effect.

A broad range of DNA lesions has been recognized in people with obesity [100,101,102]. Enhanced DNA damage was also reported in patients with obesity-related diseases such as type 2 diabetes and metabolic syndrome [103]. Moreover, body weight loss resulted in a reduction in the level of DNA damage [104,105,106,107]. The amount of DNA with oxidative damage was associated with levels of cholesterol, triglycerides and HbA1c [108]. In the present study a relationship between DNA damage and both, total cholesterol and LDL-cholesterol was also observed. Our study does have some limitations and one important limitation is a small sample size, which may be responsible for the observed lower frequency of A-allele carriers among obese than among non-obese. Moreover, we studied only women, therefore, data for men as well younger and older populations containing subjects of both genders and BMI in a wide range are needed.

In summary, the present study demonstrates the strong impact of obesity on basal DNA damage assessed by the comet assay (as % DNA in tails) and indicates that the presence of obesity abolished the protective effect of A allele on inflammation development observed in non-obese women.

References

  1. 1.

    Aseervatham GS, Sivasudha T, Jeyadevi R, Arul Ananth D (2013) Environmental factors and unhealthy lifestyle influence oxidative stress in humans—an overview. Environ Sci Pollut Res Int 20(7):4356–4369. https://doi.org/10.1007/s11356-013-1748-0

  2. 2.

    Jackson SP, Bartek J (2009) The DNA-damage response in human biology and disease. Nature 461(7267):1071–1078. https://doi.org/10.1038/nature08467

  3. 3.

    Ohnishi S, Ma N, Thanan R, Pinlaor S, Hammam O, Murata M, Kawanishi S (2013) DNA damage in inflammation-related carcinogenesis and cancer stem cells. Oxid Med Cell Longev 2013:387014. https://doi.org/10.1155/2013/387014

  4. 4.

    Cervelli T, Borghini A, Galli A, Andreassi MG (2012) DNA damage and repair in atherosclerosis: current insights and future perspectives. Int J Mol Sci 13(12):16929–16944. https://doi.org/10.3390/ijms131216929

  5. 5.

    Casorelli I, Bossa C, Bignami M (2012) DNA damage and repair in human cancer: molecular mechanisms and contribution to therapy-related leukemias. Int J Environ Res Public Health 9(8):2636–2657. https://doi.org/10.3390/ijerph9082636

  6. 6.

    Basu AK (2018) DNA damage, mutagenesis and cancer. Int J Mol Sci 19 (4). https://doi.org/10.3390/ijms19040970

  7. 7.

    Speit G, Hartmann A (2006) The comet assay: a sensitive genotoxicity test for the detection of DNA damage and repair. Methods Mol Biol 314:275–286. https://doi.org/10.1385/1-59259-973-7:275

  8. 8.

    Roy MD (2007) Approach for assessing total cellular DNA damage. Biotechniques 42(4):425–435. https://doi.org/10.2144/000112441

  9. 9.

    Azqueta A, Collins AR (2013) The essential comet assay: a comprehensive guide to measuring DNA damage and repair. Arch Toxicol 87(6):949–968. https://doi.org/10.1007/s00204-013-1070-0

  10. 10.

    Wasson GR, McKelvey-Martin VJ, Downes CS (2008) The use of the comet assay in the study of human nutrition and cancer. Mutagenesis 23(3):153–162. https://doi.org/10.1093/mutage/gen003

  11. 11.

    Moller P (2005) Genotoxicity of environmental agents assessed by the alkaline comet assay. Basic Clin Pharmacol Toxicol 96(Suppl 1):1–42

  12. 12.

    Collins AR (2004) The comet assay for DNA damage and repair: principles, applications, and limitations. Mol Biotechnol 26(3):249–261. https://doi.org/10.1385/MB:26:3:249

  13. 13.

    Garg SK, Maurer H, Reed K, Selagamsetty R (2014) Diabetes and cancer: two diseases with obesity as a common risk factor. Diabetes Obes Metab 16(2):97–110. https://doi.org/10.1111/dom.12124

  14. 14.

    Ligibel JA, Alfano CM, Courneya KS, Demark-Wahnefried W, Burger RA, Chlebowski RT, Fabian CJ, Gucalp A, Hershman DL, Hudson MM, Jones LW, Kakarala M, Ness KK, Merrill JK, Wollins DS, Hudis CA (2014) American Society of Clinical Oncology position statement on obesity and cancer. J Clin Oncol 32(31):3568–3574. https://doi.org/10.1200/JCO.2014.58.4680

  15. 15.

    Van Gaal LF, Mertens IL, De Block CE (2006) Mechanisms linking obesity with cardiovascular disease. Nature 444(7121):875–880. https://doi.org/10.1038/nature05487

  16. 16.

    Jacobs M, van Greevenbroek MM, van der Kallen CJ, Ferreira I, Blaak EE, Feskens EJ, Jansen EH, Schalkwijk CG, Stehouwer CD (2009) Low-grade inflammation can partly explain the association between the metabolic syndrome and either coronary artery disease or severity of peripheral arterial disease: the CODAM study. Eur J Clin Invest 39(6):437–444. https://doi.org/10.1111/j.1365-2362.2009.02129.x

  17. 17.

    Manna P, Jain SK (2015) Obesity, oxidative stress, adipose tissue dysfunction, and the associated health risks: causes and therapeutic strategies. Metab Syndr Relat Disord 13(10):423–444. https://doi.org/10.1089/met.2015.0095

  18. 18.

    Weisberg SP, McCann D, Desai M, Rosenbaum M, Leibel RL, Ferrante AW Jr (2003) Obesity is associated with macrophage accumulation in adipose tissue. J Clin Invest 112(12):1796–1808. https://doi.org/10.1172/JCI19246

  19. 19.

    Bukhari SA, Rajoka MI, Ibrahim Z, Jalal F, Rana SM, Nagra SA (2011) Oxidative stress elevated DNA damage and homocysteine level in normal pregnant women in a segment of Pakistani population. Mol Biol Rep 38(4):2703–2710. https://doi.org/10.1007/s11033-010-0413-7

  20. 20.

    Tomasello BMG, Galvano F, Marcella R (2011) DNA damage in normal-weight obese syndrome measured by Comet assay. Mediterr J Nutr Metab 2(4):99–104

  21. 21.

    Wlodarczyk M, Jablonowska-Lietz B, Olejarz W, Nowicka G (2018) Anthropometric and dietary factors as predictors of DNA damage in obese women. Nutrients 10(5). https://doi.org/10.3390/nu10050578

  22. 22.

    Meira LB, Bugni JM, Green SL, Lee CW, Pang B, Borenshtein D, Rickman BH, Rogers AB, Moroski-Erkul CA, McFaline JL, Schauer DB, Dedon PC, Fox JG, Samson LD (2008) DNA damage induced by chronic inflammation contributes to colon carcinogenesis in mice. J Clin Invest 118(7):2516–2525. https://doi.org/10.1172/JCI35073

  23. 23.

    Bukhari SA, Rajoka MI, Nagra SA, Rehman ZU (2010) Plasma homocysteine and DNA damage profiles in normal and obese subjects in the Pakistani population. Mol Biol Rep 37(1):289–295. https://doi.org/10.1007/s11033-009-9686-0

  24. 24.

    Setayesh T, Nersesyan A, Misik M, Ferk F, Langie S, Andrade VM, Haslberger A, Knasmuller S (2018) Impact of obesity and overweight on DNA stability: few facts and many hypotheses. Mutat Res 777:64–91. https://doi.org/10.1016/j.mrrev.2018.07.001

  25. 25.

    Hussain SP, Harris CC (2007) Inflammation and cancer: an ancient link with novel potentials. Int J Cancer 121(11):2373–2380. https://doi.org/10.1002/ijc.23173

  26. 26.

    Cerda C, Sanchez C, Climent B, Vazquez A, Iradi A, El Amrani F, Bediaga A, Saez GT (2014) Oxidative stress and DNA damage in obesity-related tumorigenesis. Adv Exp Med Biol 824:5–17. https://doi.org/10.1007/978-3-319-07320-0_2

  27. 27.

    Coussens LM, Werb Z (2002) Inflammation and cancer. Nature 420(6917):860–867. https://doi.org/10.1038/nature01322

  28. 28.

    Wang J, He Y, Yang Y, Song T, Chen N, Zhou Y (2015) Association between the TNF-alpha G-308A polymorphism and risk of ischemic heart disease: a meta-analysis. Int J Clin Exp Med 8(6):8880–8892

  29. 29.

    Zaki ME, El-Bassyouni HT, El-Gammal M, Kamal S (2015) Indicators of the metabolic syndrome in obese adolescents. Arch Med Sci 11(1):92–98. https://doi.org/10.5114/aoms.2015.49214

  30. 30.

    Kien CL, Bunn JY, Fukagawa NK, Anathy V, Matthews DE, Crain KI, Ebenstein DB, Tarleton EK, Pratley RE, Poynter ME (2015) Lipidomic evidence that lowering the typical dietary palmitate to oleate ratio in humans decreases the leukocyte production of proinflammatory cytokines and muscle expression of redox-sensitive genes. J Nutr Biochem 26(12):1599–1606. https://doi.org/10.1016/j.jnutbio.2015.07.014

  31. 31.

    Makki K, Froguel P, Wolowczuk I (2013) Adipose tissue in obesity-related inflammation and insulin resistance: cells, cytokines, and chemokines. ISRN Inflamm 2013:139239. https://doi.org/10.1155/2013/139239

  32. 32.

    Cawthorn WP, Sethi JK (2008) TNF-alpha and adipocyte biology. FEBS Lett 582(1):117–131. https://doi.org/10.1016/j.febslet.2007.11.051

  33. 33.

    Stoelcker B, Ruhland B, Hehlgans T, Bluethmann H, Luther T, Mannel DN (2000) Tumor necrosis factor induces tumor necrosis via tumor necrosis factor receptor type 1-expressing endothelial cells of the tumor vasculature. Am J Pathol 156(4):1171–1176. https://doi.org/10.1016/S0002-9440(10)64986-3

  34. 34.

    Elahi MM, Asotra K, Matata BM, Mastana SS (2009) Tumor necrosis factor alpha—308 gene locus promoter polymorphism: an analysis of association with health and disease. Biochim Biophys Acta 1792(3):163–172

  35. 35.

    Hajeer AH, Hutchinson IV (2001) Influence of TNFalpha gene polymorphisms on TNFalpha production and disease. Hum Immunol 62(11):1191–1199

  36. 36.

    Mekinian A, Tamouza R, Pavy S, Gestermann N, Ittah M, Mariette X, Miceli-Richard C (2011) Functional study of TNF-alpha promoter polymorphisms: literature review and meta-analysis. Eur Cytokine Netw 22(2):88–102. https://doi.org/10.1684/ecn.2011.0285

  37. 37.

    Kroeger KM, Carville KS, Abraham LJ (1997) The—308 tumor necrosis factor-alpha promoter polymorphism effects transcription. Mol Immunol 34(5):391–399

  38. 38.

    Koss K, Satsangi J, Fanning GC, Welsh KI, Jewell DP (2000) Cytokine (TNF alpha, LT alpha and IL-10) polymorphisms in inflammatory bowel diseases and normal controls: differential effects on production and allele frequencies. Genes Immun 1(3):185–190. https://doi.org/10.1038/sj.gene.6363657

  39. 39.

    Fernandez-Real JM, Gutierrez C, Ricart W, Casamitjana R, Fernandez-Castaner M, Vendrell J, Richart C, Soler J (1997) The TNF-alpha gene Nco I polymorphism influences the relationship among insulin resistance, percent body fat, and increased serum leptin levels. Diabetes 46(9):1468–1472

  40. 40.

    Bautista LE, Vera LM, Arenas IA, Gamarra G (2005) Independent association between inflammatory markers (C-reactive protein, interleukin-6, and TNF-alpha) and essential hypertension. J Hum Hypertens 19(2):149–154. https://doi.org/10.1038/sj.jhh.1001785

  41. 41.

    Cui G, Wang H, Li R, Zhang L, Li Z, Wang Y, Hui R, Ding H, Wang DW (2012) Polymorphism of tumor necrosis factor alpha (TNF-alpha) gene promoter, circulating TNF-alpha level, and cardiovascular risk factor for ischemic stroke. J Neuroinflammation 9:235. https://doi.org/10.1186/1742-2094-9-235

  42. 42.

    Devaraj S, O’Keefe G, Jialal I (2005) Defining the proinflammatory phenotype using high sensitive C-reactive protein levels as the biomarker. J Clin Endocrinol Metab 90(8):4549–4554. https://doi.org/10.1210/jc.2005-0069

  43. 43.

    Lio D, Scola L, Crivello A, Colonna-Romano G, Candore G, Bonafe M, Cavallone L, Marchegiani F, Olivieri F, Franceschi C, Caruso C (2003) Inflammation, genetics, and longevity: further studies on the protective effects in men of IL-10 -1082 promoter SNP and its interaction with TNF-alpha—308 promoter SNP. J Med Genet 40(4):296–299

  44. 44.

    Vikram NK, Bhatt SP, Bhushan B, Luthra K, Misra A, Poddar PK, Pandey RM, Guleria R (2011) Associations of -308G/A polymorphism of tumor necrosis factor (TNF)-alpha gene and serum TNF-alpha levels with measures of obesity, intra-abdominal and subcutaneous abdominal fat, subclinical inflammation and insulin resistance in Asian Indians in north India. Dis Markers 31(1):39–46. https://doi.org/10.3233/DMA-2011-0802

  45. 45.

    Rangel-Zuniga OA, Corina A, Lucena-Porras B, Cruz-Teno C, Gomez-Delgado F, Jimenez-Lucena R, Alcala-Diaz JF, Haro-Mariscal C, Yubero-Serrano EM, Delgado-Lista J, Lopez-Moreno J, Rodriguez-Cantalejo F, Camargo A, Tinahones FJ, Ordovas JM, Lopez-Miranda J, Perez-Martinez P (2016) TNFA gene variants related to the inflammatory status and its association with cellular aging: from the CORDIOPREV study. Exp Gerontol 83:56–62. https://doi.org/10.1016/j.exger.2016.07.015

  46. 46.

    Hoffstedt J, Eriksson P, Hellstrom L, Rossner S, Ryden M, Arner P (2000) Excessive fat accumulation is associated with the TNF alpha-308 G/A promoter polymorphism in women but not in men. Diabetologia 43(1):117–120

  47. 47.

    Phillips CM, Goumidi L, Bertrais S, Ferguson JF, Field MR, Kelly ED, Mehegan J, Peloso GM, Cupples LA, Shen J, Ordovas JM, McManus R, Hercberg S, Portugal H, Lairon D, Planells R, Roche HM (2010) Additive effect of polymorphisms in the IL-6, LTA, and TNF-{alpha} genes and plasma fatty acid level modulate risk for the metabolic syndrome and its components. J Clin Endocrinol Metab 95(3):1386–1394. https://doi.org/10.1210/jc.2009-1081

  48. 48.

    Kim HR, Lee MK, Park AJ (2006) The—308 and—238 polymorphisms of the TNF-alpha promoter gene in type 2 Diabetes Mellitus. Korean J Lab Med 26(1):58–63

  49. 49.

    Jamil K, Jayaraman A, Ahmad J, Joshi S, Yerra SK (2017) TNF-alpha—308G/A and—238G/A polymorphisms and its protein network associated with type 2 diabetes mellitus. Saudi J Biol Sci 24(6):1195–1203. https://doi.org/10.1016/j.sjbs.2016.05.012

  50. 50.

    Lee SC, Pu YB, Thomas GN, Lee ZS, Tomlinson B, Cockram CS, Critchley JA, Chan JC (2000) Tumor necrosis factor alpha gene G-308A polymorphism in the metabolic syndrome. Metabolism 49(8):1021–1024. https://doi.org/10.1053/meta.2000.7704

  51. 51.

    Walston J, Seibert M, Yen CJ, Cheskin LJ, Andersen RE (1999) Tumor necrosis factor-alpha-238 and—308 polymorphisms do not associated with traits related to obesity and insulin resistance. Diabetes 48(10):2096–2098

  52. 52.

    Kumar P, Misra S, Kumar A, Pandit AK, Chakravarty K, Prasad K (2016) Association between tumor necrosis factor-alpha (-238G/A and -308G/A) gene polymorphisms and risk of ischemic stroke: a meta-analysis. Pulse (Basel) 3(3–4):217–228. https://doi.org/10.1159/000443770

  53. 53.

    Collerton J, Martin-Ruiz C, Davies K, Hilkens CM, Isaacs J, Kolenda C, Parker C, Dunn M, Catt M, Jagger C, von Zglinicki T, Kirkwood TB (2012) Frailty and the role of inflammation, immunosenescence and cellular ageing in the very old: cross-sectional findings from the newcastle 85 + study. Mech Ageing Dev 133(6):456–466. https://doi.org/10.1016/j.mad.2012.05.005

  54. 54.

    Noren Hooten N, Ejiogu N, Zonderman AB, Evans MK (2012) Association of oxidative DNA damage and C-reactive protein in women at risk for cardiovascular disease. Arterioscler Thromb Vasc Biol 32(11):2776–2784. https://doi.org/10.1161/ATVBAHA.112.300276

  55. 55.

    Volanakis JE (2001) Human C-reactive protein: expression, structure, and function. Mol Immunol 38(2–3):189–197

  56. 56.

    Wu IC, Shiesh SC, Kuo PH, Lin XZ (2009) High oxidative stress is correlated with frailty in elderly chinese. J Am Geriatr Soc 57(9):1666–1671. https://doi.org/10.1111/j.1532-5415.2009.02392.x

  57. 57.

    Babbar N, Casero RA Jr (2006) Tumor necrosis factor-alpha increases reactive oxygen species by inducing spermine oxidase in human lung epithelial cells: a potential mechanism for inflammation-induced carcinogenesis. Cancer Res 66(23):11125–11130. https://doi.org/10.1158/0008-5472.CAN-06-3174

  58. 58.

    Jaiswal M, LaRusso NF, Burgart LJ, Gores GJ (2000) Inflammatory cytokines induce DNA damage and inhibit DNA repair in cholangiocarcinoma cells by a nitric oxide-dependent mechanism. Cancer Res 60(1):184–190

  59. 59.

    Kandhaya-Pillai R, Miro-Mur F, Alijotas-Reig J, Tchkonia T, Kirkland JL, Schwartz S (2017) TNFalpha-senescence initiates a STAT-dependent positive feedback loop, leading to a sustained interferon signature, DNA damage, and cytokine secretion. Aging (Albany NY) 9(11):2411–2435. https://doi.org/10.18632/aging.101328

  60. 60.

    Organization WH (1999) Obesity: preventing and managing the global epidemic: report of a WHO consultation; WHO technical report 894. World Health Organization: Geneva, Switzerland, World Health Organization: Geneva, Switzerland

  61. 61.

    Salinas-Santander M, Diaz-Garcia D, Rojas-Martinez A, Cantu-Salinas C, Sanchez-Dominguez C, Reyes-Lopez M, Cerda-Flores RM, Ocampo-Candiani J, Ortiz-Lopez R (2012) Tumor necrosis factor-alpha -308G/A polymorphism is associated with active vitiligo vulgaris in a northeastern Mexican population. Exp Ther Med 3(5):893–897. https://doi.org/10.3892/etm.2012.508

  62. 62.

    Singh NP, McCoy MT, Tice RR, Schneider EL (1988) A simple technique for quantitation of low levels of DNA damage in individual cells. Exp Cell Res 175(1):184–191

  63. 63.

    Wlodarczyk M, Nowicka G (2012) XPD gene rs13181 polymorphism and DNA damage in human lymphocytes. Biochem Genet 50(11–12):860–870. https://doi.org/10.1007/s10528-012-9526-0

  64. 64.

    Malik S, Wong ND, Franklin S, Pio J, Fairchild C, Chen R (2005) Cardiovascular disease in U.S. patients with metabolic syndrome, diabetes, and elevated C-reactive protein. Diabetes Care 28(3):690–693

  65. 65.

    Hotamisligil GS, Shargill NS, Spiegelman BM (1993) Adipose expression of tumor necrosis factor-alpha: direct role in obesity-linked insulin resistance. Science 259(5091):87–91

  66. 66.

    Xu H, Uysal KT, Becherer JD, Arner P, Hotamisligil GS (2002) Altered tumor necrosis factor-alpha (TNF-alpha) processing in adipocytes and increased expression of transmembrane TNF-alpha in obesity. Diabetes 51(6):1876–1883

  67. 67.

    Fain JN (2010) Release of inflammatory mediators by human adipose tissue is enhanced in obesity and primarily by the nonfat cells: a review. Mediators Inflamm 2010:513948. https://doi.org/10.1155/2010/513948

  68. 68.

    Zhang P, Wu X, Li G, He Q, Dai H, Ai C, Shi J (2017) Tumor necrosis factor-alpha gene polymorphisms and susceptibility to ischemic heart disease: a systematic review and meta-analysis. Medicine 96(14):e6569. https://doi.org/10.1097/MD.0000000000006569

  69. 69.

    Pulido-Gomez K, Hernandez-Diaz Y, Tovilla-Zarate CA, Juarez-Rojop IE, Gonzalez-Castro TB, Lopez-Narvaez ML, Alpuin-Reyes M (2016) Association of G308A and G238A polymorphisms of the TNF-alpha gene with risk of coronary heart Disease: systematic review and meta-analysis. Arch Med Res 47(7):557–572. https://doi.org/10.1016/j.arcmed.2016.11.006

  70. 70.

    Uglialoro AM, Turbay D, Pesavento PA, Delgado JC, McKenzie FE, Gribben JG, Hartl D, Yunis EJ, Goldfeld AE (1998) Identification of three new single nucleotide polymorphisms in the human tumor necrosis factor-alpha gene promoter. Tissue Antigens 52(4):359–367

  71. 71.

    Wilson AG, de Vries N, Pociot F, di Giovine FS, van der Putte LB, Duff GW (1993) An allelic polymorphism within the human tumor necrosis factor alpha promoter region is strongly associated with HLA A1, B8, and DR3 alleles. J Exp Med 177(2):557–560

  72. 72.

    Rudwaleit M, Siegert S, Yin Z, Eick J, Thiel A, Radbruch A, Sieper J, Braun J (2001) Low T cell production of TNFalpha and IFNgamma in ankylosing spondylitis: its relation to HLA-B27 and influence of the TNF-308 gene polymorphism. Ann Rheum Dis 60(1):36–42

  73. 73.

    Zeggini E, Thomson W, Kwiatkowski D, Richardson A, Ollier W, Donn R, British Paediatric Rheumatology Study Group (2002) Linkage and association studies of single-nucleotide polymorphism-tagged tumor necrosis factor haplotypes in juvenile oligoarthritis. Arthritis Rheum 46(12):3304–3311. https://doi.org/10.1002/art.10698

  74. 74.

    Sullivan KE, Wooten C, Schmeckpeper BJ, Goldman D, Petri MA (1997) A promoter polymorphism of tumor necrosis factor alpha associated with systemic lupus erythematosus in African-Americans. Arthritis Rheum 40 (12):2207–2211. https://doi.org/10.1002/1529-0131(199712)40:12%3C2207::AID-ART14%3E3.0.CO;2-Y

  75. 75.

    Birben E, Sahiner UM, Sackesen C, Erzurum S, Kalayci O (2012) Oxidative stress and antioxidant defense. World Allergy Organ J 5(1):9–19. https://doi.org/10.1097/WOX.0b013e3182439613

  76. 76.

    Hubackova S, Kucerova A, Michlits G, Kyjacova L, Reinis M, Korolov O, Bartek J, Hodny Z (2016) IFNgamma induces oxidative stress, DNA damage and tumor cell senescence via TGFbeta/SMAD signaling-dependent induction of Nox4 and suppression of ANT2. Oncogene 35(10):1236–1249. https://doi.org/10.1038/onc.2015.162

  77. 77.

    Karakasilioti I, Kamileri I, Chatzinikolaou G, Kosteas T, Vergadi E, Robinson AR, Tsamardinos I, Rozgaja TA, Siakouli S, Tsatsanis C, Niedernhofer LJ, Garinis GA (2013) DNA damage triggers a chronic autoinflammatory response, leading to fat depletion in NER progeria. Cell Metab 18(3):403–415. https://doi.org/10.1016/j.cmet.2013.08.011

  78. 78.

    Van Houten B, Hunter SE, Meyer JN (2016) Mitochondrial DNA damage induced autophagy, cell death, and disease. Front Biosci (Landmark Ed) 21:42–54

  79. 79.

    Rafieian-Kopaei M, Setorki M, Doudi M, Baradaran A, Nasri H (2014) Atherosclerosis: process, indicators, risk factors and new hopes. Int J Prev Med 5(8):927–946

  80. 80.

    Lavie CJ, Milani RV, Ventura HO (2009) Obesity and cardiovascular disease: risk factor, paradox, and impact of weight loss. J Am Coll Cardiol 53(21):1925–1932. https://doi.org/10.1016/j.jacc.2008.12.068

  81. 81.

    McDonnell DP, Park S, Goulet MT, Jasper J, Wardell SE, Chang CY, Norris JD, Guyton JR, Nelson ER (2014) Obesity, cholesterol metabolism, and breast cancer pathogenesis. Cancer Res 74(18):4976–4982. https://doi.org/10.1158/0008-5472.CAN-14-1756

  82. 82.

    Lacey B, Herrington WG, Preiss D, Lewington S, Armitage J (2017) The role of emerging risk factors in cardiovascular outcomes. Curr Atheroscler Rep 19(6):28. https://doi.org/10.1007/s11883-017-0661-2

  83. 83.

    Dickerman BA, Ahearn TU, Giovannucci E, Stampfer MJ, Nguyen PL, Mucci LA, Wilson KM (2017) Weight change, obesity and risk of prostate cancer progression among men with clinically localized prostate cancer. Int J Cancer 141(5):933–944. https://doi.org/10.1002/ijc.30803

  84. 84.

    Ferrante AW Jr (2007) Obesity-induced inflammation: a metabolic dialogue in the language of inflammation. Journal of internal medicine 262(4):408–414. https://doi.org/10.1111/j.1365-2796.2007.01852.x

  85. 85.

    Nik-Zainal S (2019) From genome integrity to cancer. Genome Med 11(1):4. https://doi.org/10.1186/s13073-019-0617-y

  86. 86.

    Tubbs A, Nussenzweig A (2017) Endogenous DNA damage as a source of genomic instability in cancer. Cell 168(4):644–656. https://doi.org/10.1016/j.cell.2017.01.002

  87. 87.

    Yang JP, Hyun MH, Yoon JM, Park MJ, Kim D, Park S (2014) Association between TNF-alpha-308 G/A gene polymorphism and gastric cancer risk: a systematic review and meta-analysis. Cytokine 70(2):104–114. https://doi.org/10.1016/j.cyto.2014.07.005

  88. 88.

    Xiao Q, Fu B, Chen P, Liu ZZ, Wang W, Ye Q (2016) Three polymorphisms of tumor necrosis factor-alpha and hepatitis B virus related hepatocellular carcinoma: a meta-analysis. Medicine 95(50):e5609. https://doi.org/10.1097/MD.0000000000005609

  89. 89.

    Winkler G, Kiss S, Keszthelyi L, Sapi Z, Ory I, Salamon F, Kovacs M, Vargha P, Szekeres O, Speer G, Karadi I, Sikter M, Kaszas E, Dworak O, Gero G, Cseh K (2003) Expression of tumor necrosis factor (TNF)-alpha protein in the subcutaneous and visceral adipose tissue in correlation with adipocyte cell volume, serum TNF-alpha, soluble serum TNF-receptor-2 concentrations and C-peptide level. Eur J Endocrinol 149(2):129–135

  90. 90.

    Di Renzo L, Sarlo F, Petramala L, Iacopino L, Monteleone G, Colica C, De Lorenzo A (2013) Association between -308 G/A TNF-alpha polymorphism and appendicular skeletal muscle mass index as a marker of sarcopenia in normal weight obese syndrome. Dis Markers 35(6):615–623. https://doi.org/10.1155/2013/983424

  91. 91.

    Yang YH, Liu YQ, Zhang L, Li H, Li XB, Ouyang Q, Zhu GY (2015) Genetic polymorphisms of the TNF-alpha-308G/A are associated with metabolic syndrome in asthmatic patients from Hebei province, China. Int J Clin Exp Pathol 8(10):13739–13746

  92. 92.

    De Luis DA, Aller R, Izaola O, Gonzalez Sagrado M, Conde R, de la Fuente B, Ovalle HF (2011) Allelic frequency of G380A polymorphism of tumor necrosis factor alpha gene and relation with cardiovascular risk factors and adipocytokines in obese patients. Nutr Hosp 26(4):711–715. https://doi.org/10.1590/S0212-16112011000400007

  93. 93.

    Rodrigues KF, Pietrani NT, Bosco AA, Campos FMF, Sandrim VC, Gomes KB (2017) IL-6, TNF-alpha, and IL-10 levels/polymorphisms and their association with type 2 diabetes mellitus and obesity in Brazilian individuals. Arch Endocrinol Metab 61(5):438–446. https://doi.org/10.1590/2359-3997000000254

  94. 94.

    Sharma R, Agrawal S, Saxena A, Sharma RK (2013) Association of IL-6, IL-10, and TNF-alpha gene polymorphism with malnutrition inflammation syndrome and survival among end stage renal disease patients. J Interferon Cytokine Res 33(7):384–391. https://doi.org/10.1089/jir.2012.0109

  95. 95.

    Gander ML, Fischer JE, Maly FE, von Kanel R (2004) Effect of the G-308A polymorphism of the tumor necrosis factor (TNF)-alpha gene promoter site on plasma levels of TNF-alpha and C-reactive protein in smokers: a cross-sectional study. BMC Cardiovasc Disord 4:17. https://doi.org/10.1186/1471-2261-4-17

  96. 96.

    Vatay A, Bene L, Kovacs A, Prohaszka Z, Szalai C, Romics L, Fekete B, Karadi I, Fust G (2003) Relationship between the tumor necrosis factor alpha polymorphism and the serum C-reactive protein levels in inflammatory bowel disease. Immunogenetics 55(4):247–252. https://doi.org/10.1007/s00251-003-0575-8

  97. 97.

    Lakka HM, Lakka TA, Rankinen T, Rice T, Rao DC, Leon AS, Skinner JS, Bouchard C (2006) The TNF-alpha G-308A polymorphism is associated with C-reactive protein levels: the HERITAGE Family Study. Vascul Pharmacol 44(5):377–383. https://doi.org/10.1016/j.vph.2006.02.002

  98. 98.

    Han CY (2016) Roles of reactive oxygen species on insulin resistance in adipose tissue. Diabetes Metab J 40(4):272–279. https://doi.org/10.4093/dmj.2016.40.4.272

  99. 99.

    Suganami T, Ogawa Y (2010) Adipose tissue macrophages: their role in adipose tissue remodeling. J Leukoc Biol 88(1):33–39. https://doi.org/10.1189/jlb.0210072

  100. 100.

    Azzara A, Pirillo C, Giovannini C, Federico G, Scarpato R (2016) Different repair kinetic of DSBs induced by mitomycin C in peripheral lymphocytes of obese and normal weight adolescents. Mutat Res 789:9–14. https://doi.org/10.1016/j.mrfmmm.2016.05.001

  101. 101.

    Donmez-Altuntas H, Sahin F, Bayram F, Bitgen N, Mert M, Guclu K, Hamurcu Z, Aribas S, Gundogan K, Diri H (2014) Evaluation of chromosomal damage, cytostasis, cytotoxicity, oxidative DNA damage and their association with body-mass index in obese subjects. Mutat Res Genet Toxicol Environ Mutagen 771:30–36. https://doi.org/10.1016/j.mrgentox.2014.06.006

  102. 102.

    Scarpato R, Verola C, Fabiani B, Bianchi V, Saggese G, Federico G (2011) Nuclear damage in peripheral lymphocytes of obese and overweight Italian children as evaluated by the gamma-H2AX focus assay and micronucleus test. FASEB J 25(2):685–693. https://doi.org/10.1096/fj.10-168427

  103. 103.

    Zaki M, Basha W, El-Bassyouni HT, El-Toukhy S, Hussein T (2018) Evaluation of DNA damage profile in obese women and its association to risk of metabolic syndrome, polycystic ovary syndrome and recurrent preeclampsia. Genes Dis 5(4):367–373. https://doi.org/10.1016/j.gendis.2018.03.001

  104. 104.

    Fejfer K, Buczko P, Niczyporuk M, Ladny JR, Hady HR, Knas M, Waszkiel D, Klimiuk A, Zalewska A, Maciejczyk M (2017) Oxidative modification of biomolecules in the nonstimulated and stimulated saliva of patients with morbid obesity treated with bariatric surgery. Biomed Res Int 2017:4923769. https://doi.org/10.1155/2017/4923769

  105. 105.

    Heilbronn LK, de Jonge L, Frisard MI, DeLany JP, Larson-Meyer DE, Rood J, Nguyen T, Martin CK, Volaufova J, Most MM, Greenway FL, Smith SR, Deutsch WA, Williamson DA, Ravussin E, Pennington CT (2006) Effect of 6-month calorie restriction on biomarkers of longevity, metabolic adaptation, and oxidative stress in overweight individuals: a randomized controlled trial. JAMA 295(13):1539–1548. https://doi.org/10.1001/jama.295.13.1539

  106. 106.

    O’Callaghan NJ, Clifton PM, Noakes M, Fenech M (2009) Weight loss in obese men is associated with increased telomere length and decreased abasic sites in rectal mucosa. Rejuvenation Res 12(3):169–176. https://doi.org/10.1089/rej.2008.0819

  107. 107.

    Soares NP, Santos AC, Costa EC, Azevedo GD, Damasceno DC, Fayh AP, Lemos TM (2016) Diet-induced weight loss reduces DNA damage and cardiometabolic risk factors in overweight/obese women with polycystic ovary syndrome. Ann Nutr Metab 68(3):220–227. https://doi.org/10.1159/000444130

  108. 108.

    Lohr M, Jensen A, Eriksen L, Gronbaek M, Loft S, Moller P (2015) Age and metabolic risk factors associated with oxidatively damaged DNA in human peripheral blood mononuclear cells. Oncotarget 6(5):2641–2653. https://doi.org/10.18632/oncotarget.3202

Download references

Acknowledgements

This work was supported by the National Science Centre (Grant No. NN404087940) and carried out with the use of the CePT infrastructure financed by the European Union- the European Regional Development Fund within the Operational Program (Innovative economy for 2007–2013).

Funding

This work was supported by the National Science Centre (Grant Nos. NN404087940 and N404042/32/0945) and carried out with the use of the CePT infrastructure financed by the European Union- the European Regional Development Fund within the Operational Program (Innovative economy for 2007–2013).

Author information

The author’s responsibilities were as follows: MW was responsible for the study concept and design, recruited patients, performed laboratory and statistical analysis, interpreted the data, and took the lead in writing the manuscript. MC created a database and contributed to statistical analysis. GN provided critical revision of the study results and intellectual content and prepared manuscript. All authors gave their final approval to the submitted manuscript.

Correspondence to Marta Włodarczyk.

Ethics declarations

Conflict of interest

Authors declared no conflict of interest.

Ethical approval

Subjects enrolled in the study were volunteers from the Outpatient Clinic of Metabolic Disorders of the National Food and Nutrition Institute. The study was conducted according to the guidelines laid down in the Declaration of Helsinki and all procedures involving human subjects were approved by the Local Ethics Committee at the National Food and Nutrition Institute.

Informed consent

Written informed consent was obtained from all of the registered volunteers. Declarations of consent in the study from all participants are available from correspondence author. Draft of the written consent form: I voluntarily and consciously agree to participate in the study. I read the information I received a copy of it. The goals and methods of the study were explained to me. I know that the study will carry out DNA analysis (carrier of genetic information) isolated from blood. I agree to perform DNA tests whose results will be used only for scientific papers. I know that participation in the study is voluntary and that I can resign from participation in the study at any time. I also declare that I have been informed that in accordance with the Act on the protection of personal data, the results of the study will not be used for other purposes than the abovementioned and that they will be kept in a place that prevents access to other people than those responsible for the above research.

Additional information

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

The original version of this article was revised.

The original publication has been updated. The names in the original publication were not in the correct order. The family name was followed by the first name. This is now corrected.

Rights and permissions

Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Włodarczyk, M., Ciebiera, M. & Nowicka, G. TNF-α G-308A genetic variants, serum CRP-hs concentration and DNA damage in obese women. Mol Biol Rep 47, 855–866 (2020) doi:10.1007/s11033-019-04764-0

Download citation

Keywords

  • DNA damage
  • Obesity
  • CRP
  • TNF gene
  • Polymorphism