Journal of Zhejiang University-SCIENCE B

, Volume 21, Issue 1, pp 42–52 | Cite as

Relationships between blood leukocyte mitochondrial DNA copy number and inflammatory cytokines in knee osteoarthritis

  • Dong Zhan
  • Aree Tanavalee
  • Saran Tantavisut
  • Srihatach Ngarmukos
  • Steven W. Edwards
  • Sittisak HonsawekEmail author


Osteoarthritis (OA) is a degenerative articular disorder manifested by cartilage destruction, subchondral sclerosis, osteophytes, and synovitis, resulting in chronic joint pain and physical disability in the elderly. The purpose of this study was to investigate mitochondrial DNA copy number (mtDNACN) and inflammatory cytokines in primary knee OA patients and healthy volunteers. A total of 204 knee OA patients and 169 age-matched healthy volunteers were recruited. Their relative blood leukocyte mtDNACN was assessed by quantitative real-time polymerase chain reaction (qRT-PCR), and ten inflammatory cytokines in their plasma were detected by multiplex immunoassay. Blood leukocyte mtDNACN in the OA group was significantly lower than that in the control group. Leukocyte mtDNACN in the control group was negatively correlated with their age (r=−0.380, P<0.0001), whereas mtDNACN in the OA group was positively correlated with their age (r=0.198, P<0.001). Plasma interleukin-4 (IL-4) and IL-6 were significantly higher in the knee OA group than in the control group. The plasma IL-6 level was positively correlated with blood leukocyte mtDNACN in the OA group (r=0.547, P=0.0014). IL-5 showed as a major factor (coefficient 0.69) in the second dimension of principle components analysis (PCA)-transformed data and was significantly higher in the OA group (P<0.001) as well as negatively correlated with mtDNACN (r=−0.577, P<0.001). These findings suggest that elevation of plasma IL-4 and IL-6 and a relative reduction in mtDNACN might be effective biomarkers for knee OA. IL-5 is a plausible factor responsible for decreasing blood leukocyte mtDNACN in knee OA patients.

Key words

Inflammatory cytokine Blood leukocyte Knee Mitochondrial DNA copy number Osteoarthritis 

膝骨关节炎患者全血白细胞线粒体 DNA 复制数量和血浆炎性细胞因子的相关性研究


目 的

本实验研究全血白细胞线粒体 DNA 复制数量 (mtDNACN)和血浆炎性细胞因子在膝骨关节炎患者中的变化和其相关性.


探讨了老年 (50~80 岁) 膝骨关节炎患者白细胞 mtDNACN 和血浆炎性细胞因子水平及二者的关系.

方 法

分别收集膝骨关节炎组和对照组血液样本并对膝关节评分 (Kellgren-Lawren grading). 使用实时定量聚合酶链反应 (qRT-PCR) 检测相对mtDNACN; 使用多重免疫分析 (multiplex immunoassay) 测定血浆中 10 种炎性细胞因子水平; 应用线性相关、 Logistic 回归和主成分分析 (PCA) 揭示骨关节炎白细胞 mtDNACN 和血浆炎性细胞因子的相关性.

结 论

血浆中白介素 4 (IL-4)、 IL-6 和全血白细胞 mtDNACN 可能是膝骨关节炎有效的生物标志物; IL-5 则对 mtDNACN 减少具有潜在的影响.


炎性细胞因子 血液白细胞 膝关节 线粒体 DNA 复制数量 骨关节炎 

CLC number



Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.



Dong ZHAN and Sittisak HONSAWEK conceived and designed the experiments, performed the experimental research, analyzed and interpreted the data, and wrote the manuscript. Aree TANAVALEE, Saran TANTAVISUT, and Srihatach NGARMUKOS collected the samples, and analyzed and interpreted the data. Steven W. EDWARDS contributed reagents, materials, and analysis tools and critically commented on the manuscript. Sittisak HONSAWEK contributed reagents, materials, and analysis tools or data, and finally edited the manuscript. All authors have read and approved the final manuscript. Therefore, all authors have full access to all the data in the study and take responsibility for the integrity and security of the data.

Compliance with ethics guidelines

Dong ZHAN, Aree TANAVALEE, Saran TANTAVISUT, Srihatach NGARMUKOS, Steven W. EDWARDS, and Sittisak HONSAWEK declare that they have no conflict of interest.

The study protocol conformed to the ethical standards outlined in the Declaration of Helsinki of 1975, as revised in 2008 (5) and was approved by the Ethical Committee on Human Research of the Faculty of Medicine, Chulalongkorn University, Thailand. All study subjects were fully informed of the study protocol and procedures prior to participating in the study. Written informed consent was obtained from all participants before any procedures were performed.


  1. Ashraf MI, Shahzad M, Shabbir A, 2015. Oxyresveratrol ameliorates allergic airway inflammation via attenuation of IL-4, IL-5, and IL-13 expression levels. Cytokine, 76(2):375–381. CrossRefGoogle Scholar
  2. Attur M, Statnikov A, Aliferis CF, et al., 2012. Inflammatory genomic and plasma biomarkers predict progression of symptomatic knee OA (SKOA). Osteoarthritis Cartilage, 20(Suppl 1):S34–S35. CrossRefGoogle Scholar
  3. Barker T, Rogers VE, Henriksen VT, et al., 2014. Serum cytokines are increased and circulating micronutrients are not altered in subjects with early compared to advanced knee osteoarthritis. Cytokine, 68(2):133–136. CrossRefGoogle Scholar
  4. Berenbaum F, 2013. Osteoarthritis as an inflammatory disease (osteoarthritis is not osteoarthrosis!). Osteoarthritis Cartilage, 21(1):16–21. CrossRefGoogle Scholar
  5. Blanco FJ, Rego I, Ruiz-Romero C, 2011. The role of mitochondria in osteoarthritis. Nat Rev Rheumatol, 7(3):161–169. CrossRefGoogle Scholar
  6. Blanco FJ, Valdes AM, Rego-Pérez I, 2018. Mitochondrial DNA variation and the pathogenesis of osteoarthritis phenotypes. Nat Rev Rheumatol, 14(6):327–340. CrossRefGoogle Scholar
  7. Chang YH, Ho KT, Lu SH, et al., 2012. Regulation of glucose/lipid metabolism and insulin sensitivity by interleukin-4. Int J Obes (Lond), 36(7):993–998. CrossRefGoogle Scholar
  8. Chawla A, Nguyen KD, Goh YPS, 2011. Macrophagemediated inflammation in metabolic disease. Nat Rev Immunol, 11(11):738–749. CrossRefGoogle Scholar
  9. Chomyn A, Attardi G, 2003. MtDNA mutations in aging and apoptosis. Biochem Biophys Res Commun, 304(3):519–529. CrossRefGoogle Scholar
  10. Dechsupa S, Singhatanadgige W, Limthongkul W, et al., 2017. Alterations of relative telomere length and mitochondrial DNA copy number from ligamentum flavum-derived cells in lumbar spinal stenosis: pilot study. Chula Med J, 61(4):497–509.Google Scholar
  11. Fang HZ, Liu XW, Shen LJ, et al., 2014. Role of mtDNA haplogroups in the prevalence of knee osteoarthritis in a southern Chinese population. Int J Mol Sci, 15(2):2646–2659. CrossRefGoogle Scholar
  12. Goekoop RJ, Kloppenburg M, Kroon HM, et al., 2010. Low innate production of interleukin-1β and interleukin-6 is associated with the absence of osteoarthritis in old age. Osteoarthritis Cartilage, 18(7):942–947. CrossRefGoogle Scholar
  13. Guo SY, Ding YJ, Li L, et al., 2015. Correlation of CD4 + CD25 + Foxp3 + Treg with the recovery of joint function after total knee replacement in rats with osteoarthritis. Genet Mol Res, 14(3):7290–7296. CrossRefGoogle Scholar
  14. Hsu FC, Kritchevsky SB, Liu YM, et al., 2009. Association between inflammatory components and physical function in the health, aging, and body composition study: a principal component analysis approach. J Gerontol A Biol Sci Med Sci, 64A(5):581–589. CrossRefGoogle Scholar
  15. Kaneko S, Satoh T, Chiba J, et al., 2000. Interleukin-6 and interleukin-8 levels in serum and synovial fluid of patients with osteoarthritis. Cytokines Cell Mol Ther, 6(2): 71–79. CrossRefGoogle Scholar
  16. Kapoor M, Martel-Pelletier J, Lajeunesse D, et al., 2011. Role of proinflammatory cytokines in the pathophysiology of osteoarthritis. Nat Rev Rheumatol, 7(1):33–42. CrossRefGoogle Scholar
  17. Kellgren JH, Lawrence JS, 1957. Radiological assessment of osteo-arthrosis. Ann Rheum Dis, 6(4):494–502. CrossRefGoogle Scholar
  18. Kopf M, le Gros G, Bachmann M, et al., 1993. Disruption of the murine IL-4 gene blocks Th2 cytokine responses. Nature, 362(6417):245–248. CrossRefGoogle Scholar
  19. Lim KS, Jeyaseelan K, Whiteman M, et al., 2005. Oxidative damage in mitochondrial DNA is not extensive. Ann N Y Acad Sci, 1042(1):210–220. CrossRefGoogle Scholar
  20. Liu SF, Kuo HC, Tseng CW, et al., 2015. Leukocyte mitochondrial DNA copy number is associated with chronic obstructive pulmonary disease. PLoS ONE, 10(9): e0138716. CrossRefGoogle Scholar
  21. Livak KJ, Schmittgen TD, 2001. Analysis of relative gene expression data using real-time quantitative PCR and the \({2^{ - {\rm{\Delta \Delta}}{C_{\rm{T}}}}}\) method. Methods, 25(4):402–408. CrossRefGoogle Scholar
  22. Livshits G, Zhai GJ, Hart DJ, et al., 2009. Interleukin-6 is a significant predictor of radiographic knee osteoarthritis: the Chingford study. Arthritis Rheum, 60(7):2037–2045. CrossRefGoogle Scholar
  23. López-Otín C, Blasco MA, Partridge L, et al., 2013. The hallmarks of aging. Cell, 153(6):1194–1217. CrossRefGoogle Scholar
  24. Mabey T, Honsawek S, Tanavalee A, et al., 2016. Plasma and synovial fluid inflammatory cytokine profiles in primary knee osteoarthritis. Biomarkers, 21(7):639–644. CrossRefGoogle Scholar
  25. Manoy P, Anomasiri W, Yuktanandana P, et al., 2018. Relationship of serum leptin and 25-hydroxyvitamin D in knee osteoarthritis patients. Chula Med J, 62(6):1037–1047. Google Scholar
  26. Meyer A, Zoll J, Charles AL, et al., 2013. Skeletal muscle mitochondrial dysfunction during chronic obstructive pulmonary disease: central actor and therapeutic target. Exp Physiol, 98(6):1063–1078. CrossRefGoogle Scholar
  27. Pedersen BK, Febbraio MA, 2008. Muscle as an endocrine organ: focus on muscle-derived interleukin-6. Physiol Rev, 88(4):1379–1406. Google Scholar
  28. Porée B, Kypriotou M, Chadjichristos C, et al., 2008. Interleukin-6 (IL-6) and/or soluble IL-6 receptor down-regulation of human type II collagen gene expression in articular chondrocytes requires a decrease of Sp1·Sp3 ratio and of the binding activity of both factors to the COL2A1 promoter. J Biol Chem, 283(8):4850–4865. CrossRefGoogle Scholar
  29. Schaap LA, Pluijm SMF, Deeg DJH, et al., 2009. Higher inflammatory marker levels in older persons: associations with 5-year change in muscle mass and muscle strength. J Gerontol A Biol Sci Med Sci, 64A(11):1183–1189. CrossRefGoogle Scholar
  30. Silvestri T, Pulsatelli L, Dolzani P, et al., 2006. Elevated serum levels of soluble interleukin-4 receptor in osteoarthritis. Osteoarthritis Cartilage, 14(7):717–719. CrossRefGoogle Scholar
  31. Smith MD, Triantafillou S, Parker A, et al., 1997. Synovial membrane inflammation and cytokine production in patients with early osteoarthritis. J Rheumatol, 24(2):365–371.PubMedGoogle Scholar
  32. Tanpaisankit M, Hongsaprabhas C, Charoenlap C, et al., 2017. High oxidative stress and decrease of mitochondrial DNA copies in musculoskeletal tumors. Chula Med J, 61(6): 771–782.Google Scholar
  33. Thomas NS, Wilkinson J, Holgate ST, 1997. The candidate region approach to the genetics of asthma and allergy. Am J Respir Crit Care Med, 156(4):S144–S151. CrossRefGoogle Scholar
  34. Vangsness CT Jr, Burke WS, Narvy SJ, et al., 2011. Human knee synovial fluid cytokines correlated with grade of knee osteoarthritis—a pilot study. Bull NYU Hosp Jt Dis, 69(2):122–127.PubMedGoogle Scholar
  35. Wu IC, Lin CC, Liu CS, et al., 2017. Interrelations between mitochondrial DNA copy number and inflammation in older adults. J Gerontol A Biol Sci Med Sci, 72(7): 937–944. CrossRefGoogle Scholar
  36. Xing JL, Chen M, Wood CG, et al., 2008. Mitochondrial DNA content: its genetic heritability and association with renal cell carcinoma. J Natl Cancer Inst, 100(15):1104–1112. CrossRefGoogle Scholar
  37. Yang YH, Bazhin AV, Werner J, et al., 2013. Reactive oxygen species in the immune system. Int Rev Immunol, 32(3): 249–270. CrossRefGoogle Scholar
  38. Yousefi S, Gold JA, Andina N, et al., 2008. Catapult-like release of mitochondrial DNA by eosinophils contributes to antibacterial defense. Nat Med, 14(9):949–953. CrossRefGoogle Scholar
  39. Zhan D, Honsawek S, 2019. Reduction of leukocyte mitochondrial DNA copy number in knee osteoarthritis. Chula Med J, 63(3):207–209. Google Scholar

Copyright information

© Zhejiang University and Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Joint PhD Program in Biomedical Sciences and Biotechnology, Faculty of MedicineChulalongkorn UniversityBangkokThailand
  2. 2.Department of Biochemistry, Osteoarthritis and Musculoskeleton Research Unit, Faculty of Medicine, King Chulalongkorn Memorial Hospital, Thai Red Cross SocietyChulalongkorn UniversityBangkokThailand
  3. 3.Department of Orthopaedics, Vinai Parkpian Orthopaedic Research Center, Faculty of Medicine, King Chulalongkorn Memorial Hospital, Thai Red Cross SocietyChulalongkorn UniversityBangkokThailand
  4. 4.Institute of Integrative BiologyUniversity of LiverpoolLiverpoolUK

Personalised recommendations