细胞因子信号抑制因子rs4969170 A/G基因多态性在慢性丙型肝炎合并胰岛素抵抗中的作用及其机制  

Study of the mechanism underlying the effect of SOCS3 rs4969170 A/G polymorphism on the occurrence of insulin resistance in chronic hepatitis C patients

作　　者：李芳[1] 郑颖颖[1] 邵翠萍[1] 范晓红[1] 王力芬[1] 霍娜[1] 陆海英[1] 吴赤红[1] 徐小元[1] 

机构地区：[1]北京大学第一医院感染疾病科,100034

出　　处：《中华肝脏病杂志》2015年第3期171-174,共4页Chinese Journal of Hepatology

基　　金：国家“十二五”重大科技专项（2012zx10002003-004-003）北京市科委科技计划重大项目（D121100003912003）

摘　　要：目的 探讨细胞因子信号抑制因子（SOCS）3rs4969170等位基因在慢性丙型肝炎合并胰岛素抵抗发生中的作用及其机制. 方法 扩增目的基因片段,通过双酶切连接法将含SOCS3rs4969170 A/G的启动子区序列克隆到pGL3-basic载体,用构建好的pGL3-SOCS3 rs4969170 A/G重组质粒转染HepG2、Huh7细胞,检测荧光素酶相对表达活性.用Western blot检测携带SOCS3rs4969170 AA/AG基因型患者外周血单个核细胞中其蛋白表达水平.采用胰岛素抵抗的稳态评估模型评估携带SOCS3 rs4969170 AA/AG基因型患者的胰岛素抵抗状态.对数据进行t检验或单因素方差分析.结果 在HepG2细胞中,表达载体pGL3-A、pGL3-G、pGL3-control的萤光素酶相对活性分别为0.12100±0.022 07、0.027 00±0.012 49、0.043 33±0.005 51,经单因素方差分析,三组差异具有统计学意义（F=48.068,P＜ 0.01）.在Huh7细胞中,三组萤光素酶相对活性分别为0.164 70±0.007 10、0.027 33±0.017 04、0.033 67±0.014 98,经单因素方差分析,三组差异也具有统计学意义（F=115.137,P＜ 0.01）.SOCS3 rs4969170 AA和AG基因型各4例慢性丙型肝炎患者的SOCS3蛋白相对表达水平分别为1.22±0.40、0.30±0.19,t=4.149,P＜0.01,差异有统计学意义.4例携带AA基因型慢性丙型肝炎患者胰岛素抵抗值为4.11±2.62,有3例发生胰岛素抵抗;而4例携带AG基因型慢性丙型肝炎患者胰岛素抵抗值为1.47±1.01,有1例发生胰岛素抵抗,t=1.881,P=0.109,差异无统计学意义. 结论 rs4969170 A/G位点的变异可能影响了SOCS3基因的转录活性及其蛋白的表达,从而干扰胰岛素信号通路,参与慢性丙型肝炎患者胰岛素抵抗的发生.Objective To study the mechanism underlying the effect of the SOCS3 rs4969170 A/G alleles on the occurrence of insulin resistance （IR） in patients with chronic hepatitis C.Methods The promoter region of the SOCS3 gene was amplified by PCR,and luciferase expression vectors were constructed and transfected into HepG2,Huh7 cell lines.The relative luciferase activity of each expression vector was assessed by the dual luciferase reporter gene assay system.Western blotting was used to detect SOCS3 protein expression in PBMCs from groups of patients with the rs4969170 AA and AG genotypes.The state of IR in eight patients was evaluated by determining their HOMA-IR.Results The pGL3-A,pGL3-G and pGL3-control vectors showed significantly different luciferase expression in the HepG2 cells （0.121 00±0.022 07,0.027 00±0.012 49 and 0.043 33±0.005 51; F =48.068,P =0.001） and in the Huh7 cell lines （0.164 70±0.007 10,0.027 33±0.017 04 and 0.033 67±0.014 98; F =115.137,P =0.001）.The expression of SOCS3 protein was significantly higher in the rs4969170 AA genotype group than in the AG genotype group （1.22±0.40 vs.0.30±0.19; t =4.149,P =0.006）.The IR index of patients with the rs4969170 AA genotype and the AG genotype was 4.11 ±2.62 and 1.47± 1.01 respectively.There were three patients with IR in the rs4969170 AA genotype group and one in the rs4969170 AG group.There was no statistically significant difference between the two genotype groups （t=1.881,P =0.109）.Conclusions The SOCS3 rs4969170 A haplotype may enhance transcriptional activity of the gene promoter to regulate gene expression,thereby increasing intracellular SOCS3 protein level and ultimately interfering with insulin signaling and causing IR in patients with chronic hepatitis C.

关 键 词：肝炎 丙型 慢性 多态性 单核苷酸 胰岛素抗药性 细胞因子信号抑制因子 

分 类 号：R512.63[医药卫生—内科学]