miR-200b通过靶向调控Ets-1对白介素-1β诱导胰岛β细胞损伤的保护作用

发布时间：2018-11-20 09:55

【摘要】：目的探讨miR-200b靶向Ets-1对白介素(IL)-1β诱导胰岛β细胞损伤的保护作用。结果体外培养大鼠胰岛β细胞株RINm5F,采用IL-1β诱导RINm5F细胞损伤,将RIN-m5F细胞分为对照组、损伤组(0.5 ng/ml IL-1β)、空转染组(0.5 ng/ml IL-1β+空载体)和过表达组(0.5 ng/ml IL-1β转染+miR-200b mimics),采用qRT-PCR检测转染48 h各组的miR-200b和Ets-1 mRNA水平,采用MTT法、FITC-Annexin V/PI双染流式细胞术及酶联免疫吸附法检测各组转染12、24、48及72 h的细胞存活率、凋亡率及胰岛素水平,检测各组细胞内谷胱甘肽过氧化物酶(GSHPx)、丙二醛(MDA)水平、总抗氧化能力(T-AOC)及超氧化物歧化酶(SOD)活性。结果与对照组相比,损伤组、空转染组的miR-200b水平降低,Ets-1 mRNA水平升高,但过表达组的miR-200b水平升高,Ets-1 mRNA水平降低(P0.05)。损伤组、空转染组转染后的存活率、上清液胰岛素水平及GSH-Px、T-AOC和SOD活性低于对照组,MDA水平和凋亡率高于对照组(P0.05);过表达组的以上指标均优于对照组(P0.05),但与对照组均有统计学差异(P0.05)。结论上调miR-200b水平对IL-1β诱导胰岛β细胞损伤有保护作用,主要表现在提高细胞存活、促进胰岛素分泌及改善氧化应激,可能与其靶向作用Ets-1有关。
[Abstract]:Objective to investigate the protective effect of miR-200b targeted Ets-1 on islet 尾 -cell injury induced by interleukin (IL)-1 尾. Results in vitro cultured rat islet 尾 cell line RINm5F, induced RINm5F cell injury by IL-1 尾. RIN-m5F cells were divided into control group (0. 5 ng/ml IL-1 尾) and control group (0. 5 ng/ml IL-1 尾). The levels of miR-200b and Ets-1 mRNA in empty transfection group (0.5 ng/ml IL-1 尾 empty vector) and overexpression group (0.5 ng/ml IL-1 尾 transfection miR-200b mimics),) were detected by qRT-PCR for 48 h. MTT assay was used. FITC-Annexin V/PI double staining flow cytometry and enzyme linked immunosorbent assay (Elisa) were used to detect the cell survival rate, apoptosis rate and insulin level at 48 and 72 hours after transfection. The malondialdehyde (MDA) level of glutathione peroxidase (GSHPx),) was detected in the cells of each group. Total antioxidant capacity (T-AOC) and superoxide dismutase (SOD) activity. Results compared with the control group, the levels of miR-200b and Ets-1 mRNA in the empty transfection group were lower than those in the control group, but the miR-200b level and Ets-1 mRNA level in the overexpression group were higher than those in the control group (P0.05). The survival rate, insulin level, GSH-Px,T-AOC and SOD activity of supernatant in the injury group and empty transfection group were lower than those in the control group, and the MDA level and apoptosis rate were higher than those in the control group (P0.05). The above indexes in the overexpression group were better than those in the control group (P0.05), but there was statistical difference between the overexpression group and the control group (P0.05). Conclusion Up-regulation of miR-200b level can protect islet 尾 cells from IL-1 尾 -induced injury, which may be related to its targeted action on Ets-1, which may be related to the increase of cell survival, insulin secretion and oxidative stress.
【作者单位】：承德医学院附属医院内分泌科;解放军总医院内分泌科;
【基金】：承德市科技支撑计划项目(No.20151049)
【分类号】：R587.1

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