CHOP介导的内质网应激在针刺干预自发性糖尿病大鼠胰岛β细胞凋亡中的影响及机制

内质网应激(ERS)启动的胰岛β细胞凋亡在2型糖尿病(T2DM)发生发展中发挥重要作用，ERS三个信号均通过CHOP启动细胞凋亡，针灸能够抑制T2DM胰岛β细胞凋亡。在既往研究基础上，本课题对CHOP介导的ERS在OLETF大鼠胰岛β细胞凋亡中的影响机制及针刺对其的作用机制进行研究。30只8周龄OLETF大鼠根据OGTT结果随机分为针刺、二甲双胍和模型组，干预8周后，胰岛β细胞CHOP、IRE1、PERK、ATF6、Bax、Bcl-2基因和蛋白分别用RT-PCR和Western blot法检测 ，TUNEL法测β细胞凋亡，胰岛素释放试验测β细胞功能，四唑盐比色法测β细胞活性，透射电镜观察β细胞超微结构，10只同周龄LETO大鼠为对照。通过分析实验数据，探讨ERS启动的CHOP通路对针刺抑制糖尿病胰岛β细胞凋亡的影响机制，以期进一步阐明针灸治疗T2DM的机理，为针灸治疗本病提供理论依据。

The incidence of type 2 diabetes(T2DM) increases seriously. Pancreatic beta-cell apoptosis initiated by endoplasmic reticulum stress(ERS) plays an important role in pathogenesis of T2DM. All of three signaling pathways of ERS initiate cell apoptosis through the CHOP pathway. Acupuncture has good curative effect on improving beta-cells function and reducing beta-cells apoptosis in subjects with T2DM. On the basis of previous studies, this study will research the influence mechanism of ERS mediated by CHOP on pancreatic beta-cells apoptosis in OLETF rats and intervention mechanism of acupuncture on it.According to OGTT test, 30 OLETF rats with 8 week-age randomly are divided into acupuncture, metformin and model group, and 10 LETO rats with same ages as control group. After 8-weeks intervention,mRNA expressions and protein contents of beta-cells' CHOP, IRE1, PERK, ATF6, Bax and Bcl-2 are detected with real time-PCR and western blot respectively. Apoptosis, function and activity of the pancreatic beta-cells are measured with TUNEL, insulin release test and tetrazolium salts colorimetric method, respectively. Beta-cells ultrastructure are observed with transmission electron microscope. To clarify further therapeutic mechanism of acupuncture on T2DM,the research will discuss the influence of CHOP pathway initiated by ERS on acupuncture inhibiting beta-cells apoptosis and improving beta-cells function in diabetic objects by analyzing experimental data.