降糖消渴颗粒含药血清对INS-1细胞FoxO1表达的影响  

Effects of Jiangtang Xiaoke Granule Contained Serum on FoxO1 Expression in INS-1 Cell

作　　者：莫芳芳[1] 刘海霞[1] 华静 赵丹丹[1] 田甜[1] 高思华[1] 

机构地区：[1]北京中医药大学,北京100029 [2]北京中医药大学第三附属医院,北京100029

出　　处：《世界中医药》2017年第12期3046-3049,3054,共5页World Chinese Medicine

基　　金：北京中医药大学中青年教师自主选题项目(2015-JYB-JSMS015)

摘　　要：目的:通过研究降糖消渴颗粒含药血清对INS-1细胞FoxO1因子表达的影响,探讨降糖消渴颗粒含药血清保护胰岛β细胞的分子机制。方法:通过慢病毒转染的方法制备FoxO1高表达INS-1稳定细胞株,以2.5μg/mL四环素干预12h诱导FoxO1高表达以成模。以10%的降糖消渴颗粒含药血清干预细胞模型24h,正常鼠血清作对照。高倍镜下观察细胞形态,MTT法计算细胞存活率,全自动生化仪检测细胞培养液中葡萄糖含量以计算葡萄糖消耗量,Elisa法检测细胞胰岛素分泌量,Western blot检测细胞中FoxO1总蛋白表达水平及其磷酸化水平,以qRT-PCR检测FoxO1mRNA表达情况。结果:高倍镜下观察细胞形态、细胞存活率均无统计学意义。降糖消渴颗粒含药血清组葡萄糖消耗量与细胞胰岛素分泌量均显著升高(P<0.01)。Western-blot结果显示FoxO1总蛋白表达无统计学意义(P>0.05),而p-FoxO1蛋白表达增加(P<0.05)。qRT-PCR结果显示FoxO1mRNA表达量稍有降低,但差异没有统计学意义(P>0.05)。结论:降糖消渴颗粒含药血清不能降低INS-1细胞FoxO1蛋白表达水平,但能促进FoxO1磷酸化,进而达到改善胰岛β细胞功能的作用。Objective： To study the molecular mechanism of Jiangtang Xiaoke Granule contained serum in protecting β-cells through studying the effects of Jiangtang Xiaoke Granule（ JTXK） contained serum on FoxO1 expression in INS-1 cell. Methods：Lentivirus transfection was used to prepare FoxO1-overexpressing INS-1 stable cell line,and the overexpression was induced by 2.5 μg/m L doxycycline for 12 hours. The cells were treated with 10% JTXK granule containing serum for 24 h. And normal rat serum was used as control. The morphological changes of cells were observed under high power microscope. MTT analysis was used to calculate cell viability. The glucose concentration in cell culture was detected by automatic biochemical analyzer. And the glucose consumption was calculated. ELISA analysis was used to examine insulin secretion. The expression levels of FoxO1 and P-FoxO1 were evaluated by western blot. And the expression levels of FoxO1 mRNA were evaluated by quantitative real-time PCR（ qRTPCR）. Results： The cells did not have significant differences in size and shape. And there was no difference in cell viability among three groups. The glucose consumption and insulin secretion were increased in JTXK group（ P〈0. 01）. At the same time,the results of western-blot and qRT-PCR showed that the expression of FoxO1 and FoxO1 mRNA had no difference（ P〈0. 05）.However,the expression of P-FoxO1 was increased（ P〈0. 05）. Conclusion： JTXK granule cannot decrease the expression of FoxO1 of INS-1 cell,but can promote phosphorylation of FoxO1,so as to improve β-cell function.

关 键 词：降糖消渴颗粒 FOXO1 INS-1细胞 2型糖尿病 

分 类 号：R285.6[医药卫生—中药学]