miR-191在香烟烟雾抽提物所致肝细胞胰岛素抵抗中的作用研究  

The effect of miR-191 on hepatic insulin resistance induced by cigarette smoke extract

作　　者：徐文超 姚杏娟[1] 许峻[2] XU Wenchao;YAO Xingjuan;XU Jun(Changzhou Center for Disease Control and Prevention,Changzhou 213022 China)

机构地区：[1]常州市疾病预防控制中心慢性非传染性疾病防治科,213022 [2]常州市第二人民医院预保科

出　　处：《中国糖尿病杂志》2019年第9期695-700,共6页Chinese Journal of Diabetes

基　　金：常州市科学技术局应用基础研究计划（CJ20179054）

摘　　要：目的探讨miR-191在香烟烟雾抽提物(CSE)所致人肝上皮L-02细胞IR中的作用。方法建立CSE诱导L-02细胞IR模型。预实验根据CSE不同处理浓度分为正常对照组(NC,CSE0μg/ml)、CSE10组(CSE10μg/ml)、CSE20组(CSE20μg/ml)、CSE40组(CSE40μg/ml)和CSE80组(CSE80μg/ml)。实验组分为正常对照组(NC)、CSE40μg/ml单独处理组(CSE)、CSE40μg/ml联合miR-191抑制剂处理组(CSE+Anti-miR-191)、CSE40μg/ml联合抑制剂空载处理组(CSE+AntimiR-NC)。ELISA检测细胞内糖原含量和葡萄糖的摄取水平;qRT-PCR检测细胞中miR-191表达;Westernblot检测蛋白水平;瞬时转染技术探讨miR-191在CSE诱导L-02细胞IR中的作用。结果CSE染毒后,L-02细胞对葡萄糖的摄取和细胞内糖原含量降低,p-IRS-1、IRS-1和p-Akt蛋白水平降低,而miR-191表达水平升高(P<0.05)。与CSE组比较,CSE+Anti-miR-191组miR-191表达降低,葡萄糖摄取、糖原含量、p-IRS-1、IRS-1和p-Akt表达升高(P<0.05)。结论miR-191通过IRS-1/Akt信号通路在CSE诱导肝细胞IR中发挥作用。Objective To investigate the effect of miR-191 in hepatic insulin resistance (IR) induced by cigarette smoke extract (CSE) in L-02 cell. Methods Human liver epithelial L-02 cell was used to establish the insulin resistance model induced by CSE. Cell models were divided into control group (NC, CSE 0 μg/ml), CSE10 group (CSE 10 μg/ml), CSE20 group (CSE 20 μg/ml), CSE40 group (CSE 40 μg/ml) and CSE80 group (CSE 80 μg/ml) according to different concentrations of CSE. The experimental group was divided into normal control group (NC group), CSE 40 μg/ml treatment group (CSE group), CSE 40 μg/ml treatment group (CSE+Anti-miR-191 group), and CSE 40 μg/ml treatment group (CSE+ Anti-miR-191 group), and CSE 40 μg/ml treatment group (CSE+ Anti-miR-NC group). ELISA kit was used to detect the glucose uptake and intracellular glycogen content. qRT-PCR was used to detect the expres- sion of miR-191 and Western blot was used to detect the protein level. Transient transfection assay was used to investigate the effect of miR-191 in CSE^induced L-02 cell IR. Results Treatment with CSE significantly decreased the levels of glucose uptake, intracellular glycogen, p-IRS-1, IRS-1 and p-Akt and signifi cantly increased the levels of miR-191 (P<0. 05). The expression of miR-191 was lower, the expression of glucose uptake, glycoge n con tent p-IRS-1, IRS-1 and p-Akt were higher in CSE+Anti-miR-191 group than in CSE group (P<0. 05). Conclusion miR-191 plays an important biological role in CSE^induced IR in hepatic L-02 cell through IRS-1/Akt signaling pathway.

关 键 词：胰岛素抵抗 香烟烟雾抽提物 微小核糖核酸191 胰岛素受体底物1 

分 类 号：R587.1[医药卫生—内分泌]