小檗碱通过PTEN/Nrf2途径抑制胰岛βTC6氧化应激及细胞凋亡  

Berberine Inhibits Oxidative Stress and Apoptosis of Pancreatic Islet βTC6via PTEN/Nrf2 Pathway

作　　者：李茉 翟佳佳 杨泽龙 张辉锋[3] 马政权 马晓瑞 李洺 马楼艳 马利 李晓苗 LI Mo;ZHAI Jia-jia;YANG Ze-long;ZHANG Hui-feng;MA Zheng-quan;MA Xiao-rui;LI Ming;MA Lou-yan;MA Li;LI Xiao-miao(Department of General Medicine,Xian Ninth Hospital,Xi'an,Shaanxi,710054,China;Department of Hepatobiliary Surgery,Xijing Hospital,Airforce Military Medical University,Xian,Shaanxi,710032,China;Department of Neurology,Xi'an Electric Power Central Hospital,Xi'an,Shaanxi,710043,China;Department of Endocrinology,Xijing Hospital,Airforce Military Medical University,Xi'an,Shaanxi,710032,China)

机构地区：[1]西安市第九医院全科医学科,陕西西安710054 [2]空军军医大学西京医院肝胆外科,陕西西安710032 [3]西安电力中心医院神经内科,陕西西安710043 [4]空军军医大学西京医院内分泌科,陕西西安710032

出　　处：《现代生物医学进展》2022年第21期4035-4042,共8页Progress in Modern Biomedicine

基　　金：西安市科技计划项目(2019114913YX004SF037(1))。

摘　　要：目的:探讨小檗碱(Berberine,BBR)在棕榈酸(palmitic acid,PA)诱导的胰岛β细胞氧化应激及凋亡中的角色及分子机制。方法:BBR和PA单独或联合处理敲低PTEN的βTC6细胞,利用MTT、Caspase-3活性检测、流式细胞术、ROS含量检测、硝基酪氨酸定量等测定各实验分组的细胞凋亡程度并比较彼此氧化应激水平,利用定量PCR以及Western blotting检测PTEN、AMPK、Nrf2的表达变化。此外,我们还评估了BBR是否可以缓解糖尿病小鼠全身炎症状态和胰岛细胞凋亡,并再次验证了BBR对糖尿病小鼠的治疗效果。结果:BBR通过降低PTEN同时升高Nrf2的表达,进而减轻PA诱导胰岛βTC6细胞ROS以及硝基酪氨酸积累,降低PA诱导性Caspase-3升高。干扰PTEN表达可以与BBR发生协同效应,即协同降低氧化应激性凋亡。经动物实验发现BBR可明显降低糖尿病小鼠血糖以及血清IL-6水平,同时在转录水平降低小鼠胰腺PTEN并上调Nrf2,TUNEL实验发现BBR可以明显抑制糖尿病小鼠胰岛细胞凋亡,而二甲双胍(Metformin, Met)未发现抑制效应。结论:BBR通过下调PTEN并上调Nrf2的表达来发挥对PA引起的βTC6细胞氧化应激以及凋亡的保护作用,而沉默PTEN可反过来与BBR形成协同保护作用。BBR与MET治疗2型糖尿病的降糖效果没有差异性,但BBR可以额外地通过PTEN/Nrf2途径发挥抗炎及抗氧化应激作用。Objective: To investigate the role and molecular mechanism of Berberine(BBR) in palmitic acid(PA)-induced oxidative stress and apoptosis of islet beta cells. Methods: The βTC6 cells with PTEN interference were treated by BBR and PA alone or in combination. MTT, Caspase-3 activity assay, flow cytometry, ROS detection, and nitrotyrosine quantification assay were used to determine the degree of cell apoptosis and the oxidative stress levels, the expression in each group. Quantitative PCR and Western blotting were adopted to detect changes of PTEN, AMPK, and Nrf2. In addition, we also evaluated whether BBR can alleviate the systematic inflammatory state and apoptosis of islet cells in diabetic mice. And the therapeutic effect of BBR on diabetic mice was reconfirmed.Results: BBR can reduce the ROS and nitrotyrosine accumulation induced by PA in βTC6 cells by decreasing PTEN and increasing Nrf2expression, and also can decrease the activity of Caspase-3. Interference of PTEN expression can have a synergistic effect with BBR, that is, the two can synergistically reduce oxidative stress induced apoptosis. It was found in animal experiments that BBR could significantly reduce the levels of blood glucose and serum IL-6 in diabetic mice, and at the same time downregulate PTEN and upregulate Nrf2.TUNEL experiments showed that BBR could significantly inhibit the apoptosis of pancreatic islet cells in diabetic mice, while Metformin(Met) had no therapeutic effect. Conclusion: BBR protects βTC6 cells from oxidative stress and apoptosis by down-regulating PTEN and up-regulating Nrf2. Silencing PTEN can in turn form a synergistic protective effect with BBR. There was no difference in the hypoglycemic effect between BBR and MET in the treatment of type 2 diabetes, but BBR could exert additional anti-inflammatory and antioxidant stress effects through the PTEN/Nrf2 pathway.

关 键 词：小檗碱 PTEN NRF2 凋亡 氧化应激 

分 类 号：R-33[医药卫生] R587.1