α-葡萄糖苷酶低表达对胰岛β细胞形态、增殖能力、炎症及氧化应激反应的影响  

Effects of low expression ofα-glucosidase on morphology,proliferation,inflammatory response and oxidative stress response of pancreaticβcells

作　　者：庞越胜 罗倩倩 蒋靓 徐逸凡 帕它木·莫合买提[1] PANG Yuesheng;LUO Qianqian;JIANG Liang;XU Yifan;Patamu Mohemaiti(School of Public health,Xinjiang Medical University,Urumqi 830017,China)

机构地区：[1]新疆医科大学公共卫生学院,乌鲁木齐830017

出　　处：《山东医药》2024年第27期17-20,共4页Shandong Medical Journal

基　　金：新疆维吾尔自治区天山雪松计划项目(2019XS03)。

摘　　要：目的观察α-葡萄糖苷酶(GAA)低表达对胰岛β细胞形态、增殖能力、炎症反应及氧化应激反应的影响,以探讨GAA在糖尿病发病机制中的潜在作用。方法分别用siRNA1、2、3、4序列干扰胰岛素瘤胰岛β细胞(βTC6细胞)中GAA的表达,结果siRNA4序列的干扰效果最好,将其作为后续实验的干扰序列。将βTC6细胞随机分为siRNA组、阴性对照组(NC组)、空白对照组(Con组)。siRNA组转染siRNA4序列,以降低GAA的表达;阴性对照组转染siRNANC序列;Con组不进行任何处理。在细胞培养24、48、72和96 h,用CCK8法观察各组细胞增殖能力;培养48 h,使用倒置显微镜观察各组细胞数目及形态,采用Western blotting法检测细胞中炎症因子IL-6和IL-1β蛋白,免疫荧光法检测细胞中活性氧(ROS)。结果与Con组和NC组相比,siRNA组在显微镜下细胞数目增加,细胞间隙变小,培养24、48、72、96 h的细胞OD值均升高,培养48 h的细胞IL-6、ROS降低(P均<0.01)。Con组与NC组上述指标比较,P均>0.05。结论GAA低表达可使胰岛β细胞形态改变,增殖能力提高,炎症反应及氧化应激反应减弱;GAA可能通过使胰岛β细胞形态改变、增强炎症反应及氧化应激反应而参与糖尿病的发病过程。Objective To observe the effects of low expression ofα-glucosidase(GAA)on morphology,proliferation,inflammatory response,and oxidative stress response of pancreaticβ-cells in order to investigate the potential role of GAA in the pathogenesis of diabetes mellitus.Methods The expression of GAA inβTC6 cells was interfered by siRNA1,2,3 and 4,respectively.As a result,the interference effect of siRNA4 was the best,and it was used as the interference sequence in subsequent experiments.βTC6 cells were randomly divided into the siRNA group,negative control group(NC group)and blank control group(Con group),respectively.Cells in the siRNA group were transfected with siRNA4 sequence to reduce the expression of GAA,cells in the negative control group were transfected with siRNANC sequence,and cells in the Con group were not treated.After 24,48,72 and 96 h of cell culture,the cell proliferation of each group was detected by CCK-8.They were cultured for 48 h,the number and morphology of cells in each group were observed by inverted microscope,the inflammatory factors IL-6 and IL-1βprotein in cells were detected by Western blotting,and the content of reactive oxygen species(ROS)in cells was detected by immunofluorescence.Results Compared with the Con and NC groups,the cells increased,the intercellular space became smaller under the microscope,the OD values of cells increased at 24,48,72,and 96 h of culture,and the levels of IL-6 and ROS decreased after 48 h of culture in the siRNA group(all P<0.01).No significant differences were found in these indicators between the Con and NC groups(all P>0.05).Conclusions Low expression of GAA can lead to alterations in cell morphology,enhanced proliferative capacity,and diminished inflammatory and oxidative stress responses of pancreaticβ-cells.GAA may participate in the pathogenesis of diabetes by changing the morphology of pancreaticβ-cells and enhancing inflammatory and oxidative stress responses.

关 键 词：Α-葡萄糖苷酶 胰岛Β细胞 βTC6细胞 细胞增殖 炎症反应 氧化应激反应 糖尿病 

分 类 号：R587.1[医药卫生—内分泌]