血管紧张素Ⅱ 1型受体自身抗体通过上调自噬诱导INS-1胰岛β细胞凋亡  

Angiotensin Ⅱ type 1 receptor autoantibodies induces INS-1 islet β-cell apoptosis by upregulation of autophagy

作　　者：李丹[1] 王瑾[1] 何金玲 冯艳金 曹竹杰 焦向英[1] LI Dan;WANG Jin;HE Jin-ling;FENG Yan-jin;CAO Zhu-jie;JIAO Xiang-ying(Department of Physiology, Shanxi Medical University, Taiyuan 030001, China)

机构地区：[1]山西医科大学生理学教研室,山西太原030001

出　　处：《中国病理生理杂志》2018年第3期474-480,共7页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.30800399);山西省重点实验室建设项目(No.2014011049-12);山西医科大学科技创新基金(No.01201406)

摘　　要：目的:本研究旨在探究血管紧张素Ⅱ1型受体自身抗体(AT1-AA)能否引起胰岛β细胞的凋亡,自噬是否参与其中以及其所发挥的作用。方法:10-6mol/L AT1-AA处理INS-1细胞24 h后,用流式细胞术、Western blot及Hoechst 33258染色检测细胞的凋亡水平;Western blot检测自噬相关蛋白LC3和beclin 1的蛋白水平。使用血管紧张素Ⅱ1型受体拮抗剂替米沙坦以及经典的自噬抑制剂3-甲基腺嘌呤(3-MA)预处理细胞1 h之后再加入AT1-AA处理24 h,检测细胞凋亡、自噬及存活率的变化情况。结果:10-6mol/L AT1-AA处理INS-1细胞24 h可明显降低细胞存活率(P<0.05)。与阴性Ig G对照组相比,AT1-AA分别处理细胞12 h、24 h和36 h后细胞凋亡水平明显增高(P<0.05);此外,LC3及beclin 1的蛋白水平也随处理时间的延长逐渐升高,且凋亡和自噬水平的升高均可被替米沙坦所阻滞。使用自噬抑制剂3-MA预处理之后,细胞的凋亡率较单独给予AT1-AA处理组明显降低(P<0.05)。结论:AT1-AA可通过血管紧张素Ⅱ1型受体上调自噬来诱导INS-1胰岛β细胞凋亡。AIM:To explore whether the angiotensin II type1receptor autoantibodies(AT1-AA)induces isletβ-cell apoptosis and whether autophagy is involved in the process.METHODS:The INS-1cells treated with AT1-AA at10-6mol/L for24h,and then the apoptosis was analyzed by flow cytometry,Western blot and Hoechst33258staining.In addition,the expression of autophagy-related proteins such as LC3and beclin1were determined by Western blot.The effects of AT1-AA on the apoptosis,autophagy and viability of INS-1cells with or without3-methyladenine(3-MA;a common autophagy inhibitor)or telmisartan(an angiotensinⅡtype1receptor blocker)pretreatment,were detected by flow cytometry,Western blot and CCK-8assay.RESULTS:Treatment with AT1-AA at10-6mol/L for24h significantly reduced the cell viability(P<0.05).Compared with the negative IgG control group,the apoptotic cells increased after incubation with AT1-AA for12h,24h and36h,respectively(P<0.05).Moreover,the protein levels of LC3and beclin1also increased gradually with the prolongation of treatment time,and the elevation of apoptosis and autophagy were blocked by telmisartan.After pretreatment with3-MA,the apoptotic rate of the cells was obviously decreased compared with the cells treated with AT1-AA alone.CONCLUSION:AT1-AA induces the apoptosis of INS-1isletβcells by upregulating autophagy via the angiotensinⅡtype1receptor pathway.

关 键 词：血管紧张素Ⅱ1型受体自身抗体 INS-1细胞 细胞凋亡 自噬 

分 类 号：R587.1[医药卫生—内分泌] R329.2[医药卫生—内科学]