MicroRNA-139-5p靶向Bcl-2调控棕榈酸诱导的胰岛β细胞凋亡  

MicroRNA-139-5p regulating palmitate-induced apoptosis of pancreaticβcells by targeting Bcl-2

作　　者：华强 孔祥 孟祥健 程锦瀚 吴涵 张妍 姚新明 Hua Qiang;Kong Xiang;Meng Xiangjian;Cheng Jinhan;Wu Han;Zhang Yan;Yao Xinming(Department of Endocrinology,The First Affiliated Hospital of Wannan Medical College,Yijishan Hospital,Wuhu 241001,Anhui,China;Anhui Education Department Key Laboratory of Research on Non-coding RNA Transformation for Major Diseases,Wannan Medical College,Wuhu 241002,Anhui,China)

机构地区：[1]皖南医学院第一附属医院,弋矶山医院内分泌科,安徽芜湖241001 [2]安徽省教育厅重大疾病非编码RNA转化研究重点实验室,皖南医学院,安徽芜湖241002

出　　处：《右江民族医学院学报》2021年第6期705-709,共5页Journal of Youjiang Medical University for Nationalities

基　　金：国家自然科学基金项目(81600645);皖南医学院自然科学基金重点项目(WK2019ZF04);安徽高校自然科学研究重点项目(KJ2020A0594);安徽省重点研究与开发计划项目(202004j07020050)。

摘　　要：目的 探索miR-139-5p在棕榈酸(palmitate,PA)诱导胰岛β细胞凋亡中的作用及机制。方法 PA孵育Min6细胞,RT-PCR检测miR-139-5p表达,Western blot检测cleaved caspase 3和Bcl-2蛋白表达,双荧光素酶报告实验验证miR-139-5p与Bcl-2之间的相互作用,GreenNuc^(TM)检测凋亡细胞。高脂饲料喂养联合STZ注射建立T2DM小鼠模型,TUNEL检测胰腺组织miR-139-5p表达。结果 过表达miR-139-5p增加cleaved caspase 3表达和Min6细胞凋亡(P<0.05)。在PA孵育的Min6细胞中miR-139-5p表达上调,而抑制miR-139-5p表达降低了cleaved caspase 3蛋白水平和Min6细胞凋亡(P <0.05)。荧光素酶报告分析表明miR-139-5p与Bcl-2相互作用,miR-139-5p过表达降低了Bcl-2表达(P <0.05),而PA诱导的Bcl-2表达下调可通过抑制miR-139-5p来恢复(P <0.05)。此外,过表达Bcl-2可拮抗miR-139-5p诱导的cleaved caspase 3蛋白上调(P <0.05),并减少凋亡的Min6细胞数量。T2DM小鼠胰腺中miR-139-5p表达增加(P <0.05),胰岛细胞凋亡增多(P <0.05)。结论 miR-139-5p抑制Bcl-2表达促进PA诱导的胰岛β细胞凋亡。Objective To explore the role and mechanism of miR-139-5p in the apoptosis of pancreaticβcells induced by palmitate(PA).Methods Min6 cells were incubated with PA.The expression of miR-139-5p was detected by RT-PCR.The expressions of cleaved caspase 3 and Bcl-2 were detected by Western blot.The dual-luciferase reporter assay was used to verify the interaction between miR-139-5p and Bcl-2.GreenNuc^(TM) was adopted to detect the apoptosis of Min6 cells.The T2DM mouse model was established by high-fat diet combined with STZ injection.TUNEL was employed to detect the expression of miR-139-5p in pancreas tissue.Results Over-expression of miR-139-5p increased the expression of cleaved caspase 3 and the apoptosis of Min6 cells(P<0.05).The expression of miR-139-5p was up-regulated in PA-incubated Min6 cells,while the inhibition of miR-139-5p expression reduced the level of cleaved caspase 3 and the apoptosis of Min6 cells.Luciferase reporter assay indicated that miR-139-5p interacted with Bcl-2.Over-expression of miR-139-5p decreased the expression of Bcl-2(P<0.05),while down-regulated expression of Bcl-2 induced by PA could be recovered by inhibiting miR-139-5p(P<0.05).In addition,over-expression of Bcl-2 antagonized the up-regulation of cleaved caspase 3 protein induced by miR-139-5p(P<0.05)and reduced the number of apoptotic Min6 cells(P<0.05).There was an increase in the expression of miR-139-5p in pancreas of T2DM mice(P<0.05),and the apoptosis of pancreatic cells also increased(P<0.05).Conclusion miR-139-5p inhibits the expression of Bcl-2 to promote the apoptosis of pancreaticβcells induced by PA.

关 键 词：糖尿病 2型 miR-139-5p 脂毒性 

分 类 号：R587.1[医药卫生—内分泌]