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Dietary PUFA Increase Apoptosis in Stomach of Patients with Dyspeptic Symptoms and Infected with H. pylori

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Lipids

Abstract

Drug-resistant strains of Helicobacter pylori and poor treatment response are the main reasons for the failure in eradicating it in patients. Polyunsaturated fatty acids (PUFA) have an inhibitory effect on bacterial growth. The aim of this study was to investigate the effect of PUFA in combination with standard triple therapy on apoptosis in H. pylori infected subjects with dyspeptic symptoms. This study was a double-blind clinical trial in which 34 H. pylori infected subjects with dyspeptic symptoms were randomly divided into two groups of 17 patients. The control group received standard triple therapy (amoxicillin, clarithromycin and omeprazole) and the experimental group received the standard therapy and PUFA for two weeks. Gene expression levels of caspase-3, BCL-2 and Bad proteins were studied with real-time PCR, while protein levels were quantified in frozen sections and using immunohistochemistry. Compared with the control group, a significant increase (p < 0.01) was observed in the expression of caspase-3 and Bad genes and a significant reduction (p < 0.05) in the expression of Bcl-2 gene. The protein level of active caspase-3 and Bad protein was significantly increased and the level of Bcl-2 protein was significantly decreased (p < 0.05). The results of this study show that oral administration of PUFA in combination with the standard triple therapy increased apoptosis in H. pylori-infected patients with dyspeptic symptoms. This increase in apoptosis may partly reduce drug resistance in these patients. Our results suggest inclusion of a dietary PUFA containing fatty acid supplement may improve treatment of patients that are refractory to the standard triple therapy.

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Abbreviations

ALA:

Alpha-linolenic acid

APAF-1:

Apoptotic protease activating factor-1

ARA:

Arachidonic acid

ASK1:

Apoptosis signal-regulating kinase 1

DAB:

3,3′-Diaminobenzidine

DHA:

Docosahexaenoic acid

EPA:

Eicosapentaenoic acid

FADD:

Fas-associated death domain

FBS:

Fast blood sugar

FasL:

Fas ligand receptor

GLA:

Gamma-linolenic acid

IAP:

Inhibitor apoptotic protein

IHC:

Immunohistochemistry

OLA:

Oleic acid

TBST:

Tris-buffered saline, 0.05% Tween20

TLR4:

Toll-like receptor-4

TNFα:

Tumor necrosis factor alpha

TRADD:

TNF receptor associated death domain

References

  1. Toller I, Neelsen K, Steger M, Hartung M, HottigerM Stucki M, Kalali B, Markus Gerhard M, Sartori A, Lopes M, Müller A (2011) Carcinogenic bacterial pathogen Helicobacter pylori triggers DNA double-strand breaks and a DNA damage response in its host cells. PNAS 108:14944–14949

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Dolatkhah H, Rahbani-Nobar M, Fattahi E, Ansari M, Mirza-Aghazadeh A, Eftekhari-Vash L, Fakhrjo A, Bahrami A (2011) Evaluation of glycemic control, gastric juice nitric oxide and oxidative stress in diabetic patients infected by Helicobacter pylori. J Med Gen Genom 3(1):1–6

    Article  CAS  Google Scholar 

  3. Dolatkhah H, Babaee S, Rahbani-Nobar M (2011) Diabetes Mellitus and Helicobacter Pylori. LAP Lambert Acad Publ

  4. Axon ATR (2007) Relationship between Helicobacter pylori gastritis, gastric cancer and gastric acid secretion. Adv Med Sci 52:55–60

    CAS  PubMed  Google Scholar 

  5. Chen DF, Hu L, Yi P, Liu WW, Fang DC, Cao H (2007) Helicobacter pylori exist in the gallbladder mucosa of patients with chronic cholecystitis. World J Gastroenterol 13:1608–1611

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Sung KC, Rhee EJ, Ryu SH, Beck SH (2005) Prevalence of Helicobacter pylori infection and its association with cardiovascular risk factors in Korean adults. Int J Cardiol 102:411–417

    Article  PubMed  Google Scholar 

  7. Ting-Jun F, Li-Hui H, Ri-Shan C, Jin L (2005) Caspase family proteases and apoptosis. Acta Biochim Biophys Sin 37(11):719–727

    Article  Google Scholar 

  8. Reed JC (2000) Mechanisms of apoptosis. Am J Pathol 157:1415–1430

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Suzuki M, Youle RJ, Tjandra N (2000) Structure of bax: coregulation of dimer formation and intracellular localization. Cell 103:645–654 (PubMed)

    Article  CAS  PubMed  Google Scholar 

  10. Shamas-Din A, Kale J, Leber B, Andrews DW (2013) Mechanisms of action of Bcl-2 family proteins. Cold Spring Harb Percept Biol 5(4):a008714

    Google Scholar 

  11. Rastogi RP, Sinha RP (2009) Molecular mechanism and pathogenicity. EXCLI J 8:155–181

    Google Scholar 

  12. Su Z, Yang Z, Xu Y, Chen Y, Yu Q (2015) Apoptosis, autophagy, necroptosis, and cancer metastasis. Mol Cancer 48:1–14

    Google Scholar 

  13. Musumeci G, Imbesi R, Szychlinska MA, Castrogiovanni P (2015) Apoptosis and skeletal muscle in aging. Open J Apoptosis 4:41–46

    Article  CAS  Google Scholar 

  14. Cullen SP, Martin SJ (2009) Caspase activation pathways: some recent progress. Cell Death Differ 16:935–938

    Article  CAS  PubMed  Google Scholar 

  15. Abel S, Riedel S, Gelderblom WC (2014) Dietary PUFA and cancer. Proc Nutr Soc 73(3):361–367

    Article  CAS  PubMed  Google Scholar 

  16. Azrad M, Turgeon C, Demark-Wahnefried W (2013) Current evidence linking polyunsaturated fatty acids with cancer risk and progression. Front Oncol 3:1–12

    Article  Google Scholar 

  17. Slagsvold JE, Pettersen CHH, Størvold GL, Follestad T, Krokan HE, Schønberg SA (2010) DHA alters expression of target proteins of cancer therapy in chemotherapy resistant SW620 colon cancer cells. Nutr Cancer 62(5):611–621

    Article  CAS  PubMed  Google Scholar 

  18. Dai J, Shen J, Pan W, Shen S, Das UN (2013) Effects of polyunsaturated fatty acids on the growth of gastric cancer cells in vitro. Lipids Health Dis 10:12–71

    Google Scholar 

  19. Mahdavi R, Nemati A, Faizi A, Amani M, Mohammadi Asl HA, Mazani M, Nagizade A, Panah Mogadam A, Pirzade A, Ghayour Nahand M (2011) Effect of ω-3 fatty acid supplementation on oxidative stress in gastric cancer patients undergoing chemotherapy. J Ardabil Univ Med Sci 11(2):166–175

    Google Scholar 

  20. Elmore S (2007) Apoptosis: a review of programmed cell death. Toxicol Pathol 35(4):495–516

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Zhou H, Chen S, Qi Y, Wang M, Jia R, Zhu D, Liu M, Liu F, Chen X, Cheng A (2015) Development and validation of a SYBR Green real-time PCR assay for rapid and quantitative detection of goose interferons and proinflammatory cytokines. Poult Sci 94(10):2382–2387

    Article  PubMed  Google Scholar 

  22. Sabzichi M, Hamishehkar H, Ramezani F, Sharifi S, Tabasinezhad M, Pirouzpanah M, Ghanbari P, Samadi N (2014) Luteolin-loaded phytosomes sensitize human breast carcinoma MDA-MB 231 cells to doxorubicin by suppressing Nrf2 mediated signalling. APJCP 15(13):5311–5316

    PubMed  Google Scholar 

  23. Livak KJ, Schmittgen TD (2011) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C (T)) method. Methods 25(4):402–408

    Article  Google Scholar 

  24. Rao X, Huang X, Zhou Z, Lin X (2013) An improvement of the 2(–delta delta CT) method for quantitative real-time polymerase chain reaction data analysis. Biostat Bioinforma Biomath 3(3):71–85

    PubMed  PubMed Central  Google Scholar 

  25. Dales JP, Plumas J, Palmerini F (2001) Correlation between apoptosis macroarray gene expression profiling and histopathological lymph node lesions. Mol Pathol 54(1):17–23

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Ummanni R, Lehnigk U, Aimmermann U, Woenckaus C, Walther R, Giebel J (2010) Immunohistochemical expression of caspase-1 and -9, uncleaved caspase-3 and -6, cleaved caspase-3 and -6 as well as Bcl-2 in benign epithelium and cancer of the prostate. Exp Ther Med 1(1):47–52

    PubMed  PubMed Central  Google Scholar 

  27. Bressenot A, Marchal S, Bezdetnaya L, Garrier J, Guillemin F, Plénat F (2009) Assessment of apoptosis by immunohistochemistry to active caspase-3, active caspase-7, or cleaved PARP in monolayer cells and spheroid and subcutaneous xenografts of human carcinoma. J Histochem Cytochem 57(4):289–300

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Walker RA (2006) Quantification of immunohistochemistry—issues concerning methods, utility and semiquantitative assessment I. Histopathology 49(406–10):97

    Google Scholar 

  29. Housman G, Byler S, Heerboth S, Lapinska K, Longacre M, Snyder N, Sarkar S (2014) Drug resistance in cancer: an overview. Cancers 6:1769–1792

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Misra V, Pandey R, Misra S-P, Dwivedi M (2014) Helicobacter pylori and gastric cancer: Indian enigma. World J Gastroenterol 20(6):1503–1509

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Manjari V, Das U (1998) Oxidant stress, anti-oxidants, nitric oxide and essential fatty acids in peptic ulcer disease. Prostaglandins Leukot Essent Fatty Acids 59(6):401–406

    Article  CAS  PubMed  Google Scholar 

  32. Das U, Vijay Kumar K, Ramanajaneyulu E, Joshi N, Dixit V (1994) Essential fatty acids and their metabolites in duodenal ulcer. Med Sci Res 22(6):423–425

    CAS  Google Scholar 

  33. Das U, Reddy D, Rao P, Radha V (1987) Essential fatty acids and peptic ulcer disease. Gut 28(7):914

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Das U, Reddy D, Rao P, Radha V (1988) Essential fatty acids and peptic ulcer disease. Gut 29(1):134

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Das U (1998) Hypothesis: cis-unsaturated fatty acids as potential anti-peptic ulcer drugs. Prostaglandins Leukot Essent Fatty Acids 58(5):377–380

    Article  CAS  PubMed  Google Scholar 

  36. Kramling HJ, Enders G, Teichmann RK, Demmel T, Merkle R, Brendel W (1987) Antigen-induced gastrin release: an immunologic mechanism of gastric antral mucosa. Adv Exp Med Biol 216A:427–429

    Article  CAS  PubMed  Google Scholar 

  37. Maeda S, Yoshida H, Mitsuno Y, Hirata Y, Ogura K, Shiratori Y, Omata M (2002) Analysis of apoptotic and antiapoptotic signalling pathways induced by Helicobacter pylori. Gut 50:771–778. doi:10.1136/gut.50.6.771

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Glauner H, Siegmund D, Motejadded H, Scheurich P, Henkler F, Janssen O, Wajant H (2002) Intracellular localization and transcriptional regulation of tumor necrosis factor (TNF) receptor-associated factor 4 (TRAF4). Eur J Biochem 269:4819–4829

    Article  CAS  PubMed  Google Scholar 

  39. Roy CT, Irwing LM, Katelaris PH, Talley NJ (1996) Do commercial kits for Helicobacter pylori infection differ in accuracy? A meta-analysis. Am J Gastroenterol 91(6):1138–1144

    Google Scholar 

  40. Yanai A, Hirata Y, Mitsuno Y, Maeda S, Shibata W, Akanuma M, Yoshida H, Kawabe T, Omata M (2003) Helicobacter pylori induces antiapoptosis through buclear factor-kappaB activation. J Infect Dis 188(11):1741–1751

    Article  CAS  PubMed  Google Scholar 

  41. Murata-Kamiya N, Kurashima Y, Teishikata Y, Yamahashi Y, Saito Y, Higashi H, Aburatani H, Akiyama T, Peek RM Jr, Azuma T, Hatakeyama M (2007) Helicobacter pylori CagA interacts with E-cadherin and deregulates the b-catenin signal that promotes intestinal transdifferentiation in gastric epithelial cells. Oncogene 26:4617–4626

    Article  CAS  PubMed  Google Scholar 

  42. Lee S, Kim SM, Lee RT (2013) Thioredoxin and thioredoxin target proteins: from molecular mechanisms to functional significance. Antioxid Redox Signal 18(10):1165–1207

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Matsuzawa A, Saegusa K, Noguchi T, Sadamitsu C, Nishitoh H, Nagai S, Koyasu S, Matsumoto K, Takeda K, Ichijo H (2005) ROS-dependent activation of the TRAF6-ASK1-p38 pathway is selectively required for TLR4-mediated innate immunity. Nat Immunol 6:587–592

    Article  CAS  PubMed  Google Scholar 

  44. Ichijo H, Nishida E, Irie K, Dijke P, Saitoh M, Moriguchi T, Takagi M, Matsumoto K, Miyazono K, Gotoh Y (1997) Induction of apoptosis by ASK1, a mammalian MAPKKK that activates SAPK/JNK and p38 signaling pathways. Science 275(5296):90–94

    Article  CAS  PubMed  Google Scholar 

  45. Correia M, Michel V, Matos A, Carvalho P, Oliveira MJ, Ferreira RM, Dillies M-A, Huerre M, Seruca R, Figueiredo C, Machado JC, Touati E (2012) Docosahexaenoic acid inhibits Helicobacter pylori growth in vitro and mice gastric mucosa colonization. PLoS One 7:1–9

    Google Scholar 

  46. Duggan AEC, Atherton J, Cockayne A (1997) Clarification of the link between polyunsaturated fatty acids and Helicobacter pylori-associated duodenal ulcer disease: a dietary intervention study. Br J Nutr 78:515–522

    Article  CAS  PubMed  Google Scholar 

  47. Schley PD, Brindley DN, Field CJ (2007) (n-3) PUFA alter raft lipid composition and decrease epidermal growth factor receptor levels in lipid rafts of human breast cancer cells. J Nutr 1373:548–553

    Google Scholar 

  48. Lee SE, Lim JW, Kim JM, Kim H (2014) Anti-inflammatory mechanism of polyunsaturated fatty acids in Helicobacter pylori-infected gastric epithelial cells. Mediat Inflamm 2014:1–13

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Acknowledgements

This project required extensive research and dedication from many individuals. Therefore we would like to extend our sincere gratitude to all of them and to the subjects for their participation in this study. First, we are thankful to the Tabriz Liver and Gastrointestinal Disease Research Center and Tehran Islamic Azad University Science and Research Branch for their financial and logistical support and for providing necessary guidance concerning project implementation. We are also grateful to the Tabriz Endoscopy Department of Imam Rewa and Shahid-Madani Hospital for provision of expertise and technical support in the implementation. Without their superior knowledge and experience, this study would lack quality of outcomes, and thus their support was essential.

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Authors and Affiliations

  1. Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran

    Rasoul Sharifi, Akram Eidi & Zahra Noormohammadi

  2. Department of Clinical Biochemistry and Laboratories Medicine, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, East-Azarbaijan, Iran

    Mohammad Nouri

  3. Liver and Gastrointestinal Disease Research Center, Tabriz University of Medical Sciences, Tabriz, East-Azarbaijan, Iran

    Homayun Dolatkhah

  4. Department of Internal Medicine, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, East-Azarbaijan, Iran

    Masoud Shirmohammadi

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  1. Rasoul Sharifi
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Correspondence to Mohammad Nouri.

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Sharifi, R., Nouri, M., Eidi, A. et al. Dietary PUFA Increase Apoptosis in Stomach of Patients with Dyspeptic Symptoms and Infected with H. pylori . Lipids 52, 549–558 (2017). https://doi.org/10.1007/s11745-017-4257-y

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