文章摘要
古联,黄静妍,苏莉,梁宝云,杨俊威,吴艳丽,陈子文,陈清,唐农.Toll样受体4基因多态性与汉族男性中风证型关联性研究[J].南京中医药大学学报,2016,32(6):513-518.
Toll样受体4基因多态性与汉族男性中风证型关联性研究
Influences of Toll-like Receptor-4 Gene Polymorphism on Occurance of Stroke Due to Qi Deficiency and Blood Stasis in Han Chinese Men
  
DOI:
中文关键词: 中风  TLR4  单核苷酸多态性  mRNA  炎性细胞因子
英文关键词: stroke  TLR4  single nucleotide polymorphism  mRNA  inflammatory cytokines
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作者单位
古联,黄静妍,苏莉,梁宝云,杨俊威,吴艳丽,陈子文,陈清,唐农 1.广西中医药大学第一附属医院广西 南宁 530023
2.广西中医药大学研究生学院广西 南宁 530001
3.广西医科大学公共卫生学院广西 南宁 530021 
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中文摘要:
      目的 探讨中国南方汉族人Toll 样受体4基因(TLR4)多态性rs1927914、rs10759932和rs11536889与中风证型(风痰瘀阻证、气虚血瘀证)易感性、基因表达水平、细胞因子及血脂代谢的相关性。方法 采用病例对照研究方法,将559例中风患者按中医辨证分气虚血瘀型组(248例)及风痰瘀阻型组(311例),另设一性别、年龄相匹配健康对照组(605例)。所有研究对象均来自汉族。采用Sequenom Mass ARRAY 技术对TLR4基因的rs1927914、rs10759932和rs11536889进行基因分型检测,实时荧光定量PCR(qPCR)检测TLR4信使核糖核酸(mRNA)表达水平,酶联免疫吸附测定(ELISA)检测白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、白细胞介素-8(IL-8)、白细胞介素-12(IL-12)、肿瘤坏死因子-α(TNF-α)水平,HITACHI日立7600全自动生化分析仪测定总胆固醇(TC)、甘油三酯(TG)、高密度脂蛋白胆固醇(HDL)、低密度脂蛋白胆固醇(LDL)水平。结果 rs10759932的等位基因频率在中风气虚血瘀型男性病例与男性对照组之间的分布存在统计学差异(P<0.05)。rs1927914(显性模型:OR=0.65, 95% CI=0.43-0.96, Padj=0.031)、rs10759932(显性模型:OR=0.64, 95% CI=0.41-0.99, Padj=0.046)位点多态性与男性中风气虚血瘀证易感性显著关联。携带rs10759932位点TT基因型的中风气虚血瘀型患者的IL-8血清水平显著低于CC+CT者(P<0.01)。rs11536889多态性与中风气虚血瘀型男性患者的TG水平 (加性模型:Padj=0.021, 显性模型:Padj=0.013)、HDL (显性模型:Padj=0.032)显著关联。结论 TLR4基因多态性影响汉族男性中风气虚血瘀证的发生及炎性反应和血脂代谢病理过程。
英文摘要:
      OBJECTIVE To explore the correlation between polymorphisms in the toll-like receptor 4 (rs1927914, rs10759932 and rs11536889) and susceptivity, gene expression, cytokines and blood lipid metabolism in southern Han Chinese with stroke due to wind-phlegm obstruction, qi deficiency and blood stasis. METHODS 559 stroke cases were divided into two groups based on case-control study method, with 248 cases in the qi deficiency and blood stasis group, 311 cases in wind-phlegm obstruction group. In addition, another 605 cases were involved as the healthy control group with the corresponding gender and age. All the subjects were from the Han nationality. Sequenom MassARRAY technology was adopted for testing and genotyping of rs1927914, rs10759932 and rs11536889 in the toll-like receptor 4. Real-time fluorescence quantification(qPCR) was applied for examine mRNA expression. Enzyme linked immunosorbent assay(ELISA) was used to detect the level of interleukin-1β(IL-1β), interleukin-6 (IL-6), interleukin-8 (IL-8), interleukin-12(IL-12) and tumor necrosis factor-α(TNF-α). HITACHI 7600 automatic biochemical analyzer was applied to test the total cholesterol(TC), triglyceride(TG), high density lipoprotein cholesterol(HDL) and low density lipoprotein cholesterol(LDL). RESULTS There existed a statistical difference between the male cases of the qi deficiency and blood stasis group and male cases of the control group in the distribution of gene frequency of rs10759932(P<0.05). The locus polymorphism of both rs1927914 (dominant model: OR=0.65, 95%CI=0.43-0.96, Padj=0.031) and rs10759932 (dominant model: OR=0.64, 95%CI=0.41-0.99, Padj=0.046) were closely related with susceptivity of stroke male of qi deficiency and blood stasis type. The IL-8 level of male stroke patients of qi deficiency and blood stasis type with the rs10759932 TT genotype was much lower than those with CC+CT(P<0.01). Rs11536889 polymorphism was notablely related to TG level(additive model: Padj=0.021, dominant model: Padj=0.013) and HDL level(dominant model: Padj=0.032) in stroke patients of qi deficiency and blood stasis type. CONCLUSION TLR4 polymorphisms exert certain influence on the occurance of stroke due to qi deficiency and blood stasis in Han Chinese men and they have been involved in the inflammatory reaction and blood lipid metabolism.
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