升清降糖合剂对氧化损伤胰岛β细胞Bcl2BaxAkt表达影响.docVIP

升清降糖合剂对氧化损伤胰岛β细胞Bcl2BaxAkt表达影响 　　摘 要：目的：研究升清降糖合剂（SQ）对氧化损伤胰岛β细胞凋亡的保护作用，探讨其对凋亡蛋白Bcl-2和Bax、Akt蛋白表达的影响。方法：H2O2诱导胰岛细胞氧化损伤模型，流式细胞术测定胰岛细胞凋亡率，以荧光免疫法检测细胞凋亡蛋白Bcl-2和Bax的表达，免疫印迹测定细胞Akt磷酸化表达。结果：模型组细胞凋亡率增高，达（35.18±2.41）%，与正常组比较有差异（P0.01），与SQ保护组比较有差异（P0.01）。模型组细胞内Bax荧光表达较正常组增加（P0.05）；SQ保护组Bcl-2荧光表达上升，Bax荧光表达下降，与模型组比较，均有统计学差异（P0.05）。对于胰岛细胞瘤株RINm5F，模型组磷酸化Akt蛋白表达减少，与正常组比较有统计学差异（P0.05），与保护组比较有统计学差异（P0.01）；对于原代胰岛，模型组磷酸化Akt蛋白减少，与正常组比较有统计学差异（P0.05），与保护组比较有统计学差异（P0.05）。结论：升清降糖合剂可降低氧化损伤胰岛β细胞凋亡率，上调Bcl-2，抑制Bax蛋白表达，促进Akt磷酸化。升降糖合剂抗氧化和抗凋亡机制与调节凋亡蛋白Bcl-2和Bax及PI3K-Akt通路有关。 　　关键词：升清降糖合剂；糖尿病；细胞凋亡；氧化损伤；Bcl-2；Bax；丝/苏氨酸蛋白激酶；半仿生提取法 　　 中图分类号：R587.1文献标识码：A文章编号：1673-7717(2011)12-2692-03 　　Effect of Shengqing Jiangtang Decoction on Expression of Bcl-2， 　　Bax，Akt in Beta Cells with Oxidative Damage 　　NI Xiaofen1，HU Zhen2，ZHENG Chao2，XU Xiaofeng1 　　(1.First Affiliated Hospital of Wenzhou Medical College，Wenzhou 325000，Zhejiang，China； 　　2. Second Affiliated Hospital of Wenzhou Medical College，Wenzhou 325003，Zhejiang，China) 　　 Abstract:It is a study on protective mechanism of Shengqing Jiangtang decoction (SQ) on pancreatic beta-cells with oxidative damage. We made the oxidative damage of beta-cells by H2O2 and interfered with SQ, we assayed the cell apotosis rate，the expression levels of Bcl-2，Bax and phosphorylated Akt (pAkt) by flow cytometry, fluroimmuno-assay and Western Blot method, respectively. We found the apotosis rate of model group (35.18±2.41)％ had statistical defference compared with protection group (19.56±1.08)％,P0.01. Bcl-2 expression increased and Bax expression decreased in protection group, which had a difference compared with model group(P0.05), respectively. We found pAkt protein expressed in both RINm5F and primary rat pancreatic islets. In each object, expression of pAkt declined in model group, which had a statistical difference compared with negative group; pAkt increased in protection group, which had a statistical difference compared with model group, respectively. 　　 Ke