肿瘤坏死因子受体相关因子6基因多态性对中风风痰瘀阻证发生风险及炎性反应过程的影响
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摘要
目的探讨肿瘤坏死因子受体相关因子6(TRAF6)基因多态性与中风不同证型遗传易感性以及炎性反应相关数量性状的关联。方法本研究采用病例-对照设计,纳入中风风痰瘀阻证者311例、气虚血瘀证者284例、性别年龄相匹配的对照组605例。采用实时荧光定量PCR(RTPCR)方法检测了TRAF6 mRNA表达水平,运用Sequenom Mass ARRAY i PLEX平台对TRAF6基因rs5030411、rs5030416两个多态性位点进行基因分型,采用酶联免疫吸附实验法(ELISA)测定炎性细胞因子血清水平。用PLINK、SPSS软件进行统计学分析。结果中风风痰瘀阻证、气虚血瘀证患者的TRAF6 mRNA表达水平均显著高于对照组(P<0.050)。TRAF6基因rs5030416多态性与中风风痰瘀阻证的易感性存在统计学关联(P=0.030),rs5030411多态性与中风易感性无统计学关联(P>0.050)。携带rs5030411 CT基因型的风痰瘀阻证患者的白细胞介素-1b(interleukin-1b,IL-1b)、白细胞介素-6(interleukin-6,IL-6)血清水平显著高于其他基因型携带者。风痰瘀阻证患者的IL-1b、IL-6血清水平显著高于对照组(P<0.050)。结论 TRAF6基因rs5030416多态性可能影响中风风痰瘀阻证的发生,而rs5030411多态性可能参与调节风痰瘀阻证的炎性反应过程。
Objective To investigate the relationship between tumor necrosis factor receptor-associated factor 6( TRAF6) gene polymorphisms and susceptibility to stroke with diverse TCM patterns and inflammation-related quantitative characters. Methods A case-control design was applied in this study. 311 stroke cases with the pattern of wind-phlegm stasis and 284 cases with qi-deficiency blood-stasis pattern were included in the case group,and 605 gender-matched,and age-matched cases were enrolled in the control group. The expression level of TRAF6 mRNA was determined by using real-time PCR( RTPCR),and two single nucleotide polymorphisms( SNP) of the TRAF6 gene,rs5030411 and rs5030416,were genotyped with Sequenom Mass ARRAY i PLEX platform. Enzyme-linked immuno sorbent assay( ELISA) was applied to detect the serum levels of inflammatory cytokines. PLINK and SPSS software were used for statistical analysis. Results The TRAF6 mRNA expression level in patients with windphlegm stasis pattern or qi-deficiency blood-stasis pattern were significantly higher than those in the control group( P < 0. 050). Rs5030416 polymorphism has a close association with susceptibility to stroke with wind-phlegm stasis pattern( P = 0. 030). No relationship was found between rs5030411 polymorphism and stroke susceptibility( P > 0. 050). Patients carrying rs5030411-CT genotype in wind-phlegm stasis pattern group had remarkably higher interleukin-1 b( IL-1 b) and interleukin-6( IL-6) levels than other genotype carriers. Moreover,compared with control group,the levels of IL-1 b and IL-6 were significantly higher in wind-phlegm stasis pattern group( P < 0. 05). Conclusion TRAF6 gene polymorphism rs5030416 may play a role in the occurrence of wind-phlegm stasis pattern,and rs5030411 polymorphism may engage in the inflammatory reaction in wind-phlegm stasis pattern.
Objective To investigate the relationship between tumor necrosis factor receptor-associated factor 6( TRAF6) gene polymorphisms and susceptibility to stroke with diverse TCM patterns and inflammation-related quantitative characters. Methods A case-control design was applied in this study. 311 stroke cases with the pattern of wind-phlegm stasis and 284 cases with qi-deficiency blood-stasis pattern were included in the case group,and 605 gender-matched,and age-matched cases were enrolled in the control group. The expression level of TRAF6 mRNA was determined by using real-time PCR( RTPCR),and two single nucleotide polymorphisms( SNP) of the TRAF6 gene,rs5030411 and rs5030416,were genotyped with Sequenom Mass ARRAY i PLEX platform. Enzyme-linked immuno sorbent assay( ELISA) was applied to detect the serum levels of inflammatory cytokines. PLINK and SPSS software were used for statistical analysis. Results The TRAF6 mRNA expression level in patients with windphlegm stasis pattern or qi-deficiency blood-stasis pattern were significantly higher than those in the control group( P < 0. 050). Rs5030416 polymorphism has a close association with susceptibility to stroke with wind-phlegm stasis pattern( P = 0. 030). No relationship was found between rs5030411 polymorphism and stroke susceptibility( P > 0. 050). Patients carrying rs5030411-CT genotype in wind-phlegm stasis pattern group had remarkably higher interleukin-1 b( IL-1 b) and interleukin-6( IL-6) levels than other genotype carriers. Moreover,compared with control group,the levels of IL-1 b and IL-6 were significantly higher in wind-phlegm stasis pattern group( P < 0. 05). Conclusion TRAF6 gene polymorphism rs5030416 may play a role in the occurrence of wind-phlegm stasis pattern,and rs5030411 polymorphism may engage in the inflammatory reaction in wind-phlegm stasis pattern.
引文
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[2]WANG YC,LIN S,YANG QW.Toll-like receptors in cerebral ischemic inflammatory injury[J].Journal of Neuroinflammation,2011,8(8):134.
[3]CASO JR,PRADILLO JM,HURTADO O,et al.Toll-like receptor 4 is involved in brain damage and inflammation after experimental stroke[J].Circulation,2007,115(12):1599-1608.
[4]刁殿军,张增瑞.缺血性中风病机探析[J].天津中医药,2012,29(4):359-360.DIAO DJ,ZHANG ZR.Study of pathogenesis in ischemic stroke[J].Tianjin Journal of Traditional Chinese Medicine,2012,29(4):359-360.
[5]陈鹏.基因多态性与中医辨证关系研究的回顾与思考[J].中国中西医结合杂志,2003,23(11):852-854.CHEN P.Retrospect and thought about the relationship between gene polymorphism and syndrome differentiation in traditional Chinese medicine[J].Chinese Journal of Integrated Traditional and Western Medicine,2003,23(11):852-854.
[6]涂晋文,董梦久,邵卫,等.血管紧张素转化酶基因ACE(I/D)多态性与缺血性脑卒中中医证候相关性分析[J].浙江中医杂志,2009,44(3):162-164.TU JW,DONG MJ,SHAO W,et al.Correlation analysis between ACE(I/D)gene polymorphism and TCM syndrome of ischemic stroke[J].Zhejiang Journal of Traditional Chinese Medicine,2009,44(3):162-164.
[7]胡木林,王忠,李涛,等.缺血性中风血瘀证患者MTHFR基因多态性研究[J].中国中医药信息杂志,2009,16(3):16-18.HU ML,WANG Z,LI T,et al.Study on MTHFR C677T polymorphism in ischemic stroke with blood stasis syndrome[J].Chinese Journal of Information on Traditional Chinese Medicine,2009,16(3):16-18.
[8]WU D,LEE YCG,LIU HC,et al.Identification of TLR downstream pathways in stroke patients[J].Clinical Biochemistry,2013,46(12):1058-1064.
[9]AKIRA S,TAKEDA K.Toll-like receptor signaling[J].Nature Reviews Immunology,2004,4(7):499-511.
[10]MOGHIMPOUR BF,VALLEJO JG,REZAEI N.Toll-like receptor signaling pathways in cardiovascular diseases:challenges and opportunities[J].International Reviews of Immunology,2012,31(5):379-395.
[11]YUAN P,LIU ZQ,LIU MH,et al.Up-regulated tumor necrosis factor-associated factor 6 level is correlated with apoptosis in the tat cerebral ischemia and reperfusion[J].Neurological Sciences,2013,34(7):1133-1138.
[12]LAI HM,LI XM,YANG YN,et al.Genetic variation in NFKB1 and NFKBIA and susceptibility to coronary artery disease in a Chinese Uygur population[J].PLo S One,2015,10(6):e0129144.
[13]SARTORIUS T,STAIGER H,KETTERER C,et al.Association of common genetic variants in the MAP4K4 locus with prediabetic traits in humans[J].PLo S One,2012,7(10):e47647.
[14]李澎涛,王永炎,黄启福.“毒损脑络”病机假说的形成及其理论与实践意义[J].北京中医药大学学报,2001,24(1):1-6.LI PT,WANG YY,HUANG QF.The hypothesis of the injury of brain collaterals by toxins and its theoretical and practical significance[J].Journal of Beijing University of Traditional Chinese Medicine,2001,24(1):1-6.
[15]仓志兰,姜亚军.急性脑梗死中医证型与细胞因子的关系研究[J].四川中医,2006,24(7):19-20.CANG ZL,JIANG YJ.Association between traditional Chinese medicine syndromes and cytokines in patients with acute cerebral infarction[J].Journal of Sichuan of Traditional Chinese Medicine,2006,24(7):19-20.