细胞外信号调节激酶1/2在白介素-1β抑制胰岛β细胞葡萄糖刺激下胰岛素分泌中的作用  

Effects of ERK1/2 signal transduction pathway on the GSIS inhibited by IL-1β in βTC-6 cells

作　　者：李超[1,2] 合湫[1] 王敏[3] 苏恒[1] 李清竹[1] 沈涛[4] 薛元明[1] 

机构地区：[1]云南省第一人民医院内分泌代谢科,云南省糖尿病研究中心,昆明650032 [2]玉溪市第一人民医院急诊科 [3]云南大学现代分析测试中心 [4]云南省第一人民医院基础研究所,昆明650032

出　　处：《中国糖尿病杂志》2015年第6期558-561,共4页Chinese Journal of Diabetes

基　　金：云南省中青年学术技术带头人后备人才项目(2007PY01-32)

摘　　要：目的探讨细胞外信号调节激酶1/2(ERK1/2)信号转导通路在人IL-1β抑制小鼠胰岛素瘤胰岛β细胞(βTC-6)葡萄糖刺激下胰岛素分泌反应(GSIS)中的作用。方法 (1)βTC-6细胞分别于含不同浓度(0、1.38、5.5mmol/L)葡萄糖的KRBH缓冲液中孵育60min,放射免疫法检测细胞上清液中胰岛素浓度;(2)βTC-6细胞分别于含不同浓度(0、1.38、5.5 mmol/L)葡萄糖的KRBH缓冲液中孵育5min,蛋白质免疫印迹法检测细胞蛋白中磷酸化ERK1/2及β-actin的水平;(3)βTC-6细胞加入IL-1β(0.15、1.5、15ng/ml)培养24h后于含1.38mmol/L葡萄糖的KRBH缓冲液中孵育60min,检测上清液中胰岛素浓度;(4)βTC-6细胞加入IL-1β(0.15、1.5、15ng/ml)培养24h后于含1.38mmol/L葡萄糖的KRBH缓冲液中孵育5min,检测细胞蛋白中磷酸化ERK1/2及β-actin的水平。结果 (1)βTC-6细胞在1.38mmol/L葡萄糖刺激时胰岛素分泌及ERK1/2磷酸化水平均达到高峰;(2)IL-1β可抑制βTC-6细胞葡萄糖刺激下的ERK1/2磷酸化及胰岛素分泌,作用与剂量呈正相关。结论 ERK1/2信号转导通路可能在IL-1β抑制βTC-6细胞葡萄糖刺激下的胰岛素分泌反应中发挥重要作用。Objective To investigate effects of extracellular signal-regulated kinase 1/2（ERK1/2）signal transduction pathway on glucose stimulated insulin secretion（GSIS）inhibited by IL-1βinβTC-6cells.Methods（1）βTC-6cells were incubated in Krebs-Ringer bicarbonate（KRB）buffer containing 0,1.38 or 5.5mmol/L glucose for 60 mins.Supernatants were collected for insulin assays by an insulin RIA kit.（2）βTC-6cells were incubated in KRB buffer containing 0,1.38 or 5.5mmol/L glucose for 5mins.At the end of the incubation,cells were suspended with lysis buffer and cell lysate was used for Western blot to test the levels of phosphorylated ERK1/2andβ-actin’s.（3）βTC-6cells were incubated in DMEM containing IL-1β（0.15,1.5,15ng/ml）for 24 hours.DMEM was then replaced with fresh KRB buffer containing 1.38mmol/L glucose for 60 mins.Supernatants were collected for insulin assays.（4）βTC-6cells were incubated in DMEM containing IL-1β（0.15,1.5,15ng/ml）for 24 hours.DMEM was then replaced with KRB buffer containing 1.38mmol/L glucose for 5mins.At the end of the incubation,cells were suspended with lysis buffer and cell lysate was used for Western blot.Results（1）The insulin level and ERK1/2phosphorylation reached a peak in response to 1.38 mmol/L glucose stimulation inβTC-6cells.（2）IL-1βcan inhibit ERK1/2phosphorylation and insulin secretion stimulated by glucose stimulation in a dose-dependent manner inβTC-6cells.Conclusion ERK1/2signal transduction pathway activation may have an important effect on the GSIS inhibited by IL-1βinβTC-6cells.

关 键 词：细胞外信号调节激酶1/2 信号转导通路 小鼠胰岛素瘤胰岛β细胞 糖尿病 

分 类 号：R587.1[医药卫生—内分泌]