首页|期刊导航|山东医药|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

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OACSTPCD

Effects of low expression of α-glucosidase on morphology,proliferation,inflammatory response and oxidative stress response of pancreatic β cells

中文摘要英文摘要

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 of α-glucosidase(GAA)on morphology,prolifera-tion,inflammatory response,and oxidative stress response of pancreatic β-cells in order to investigate the potential role of GAA in the pathogenesis of diabetes mellitus.Methods The expression of GAA in βTC6 cells was interfered by siR-NA1,2,3 and 4,respectively.As a result,the interference effect of siRNA4 was the best,and it was used as the interfer-ence 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 siR-NA4 sequence to reduce the expression of GAA,cells in the negative control group were transfected with siRNANC se-quence,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-1β protein in cells were detected by Western blot-ting,and the content of reactive oxygen species(ROS)in cells was detected by immunofluorescence.Results Com-pared 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 cul-ture 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 prolif-erative capacity,and diminished inflammatory and oxidative stress responses of pancreatic β-cells.GAA may participate in the pathogenesis of diabetes by changing the morphology of pancreatic β-cells and enhancing inflammatory and oxidative stress responses.

庞越胜;罗倩倩;蒋靓;徐逸凡;帕它木·莫合买提

新疆医科大学公共卫生学院,乌鲁木齐 830017

临床医学

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

α-glucosidasepancreatic β-cellsβTC6 cellscell proliferationinflammatory responseoxidative stress responsediabetes mellitus

《山东医药》 2024 (027)

17-20 / 4

新疆维吾尔自治区天山雪松计划项目(2019XS03).

10.3969/j.issn.1002-266X.2024.27.004

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