Abstract:

Objective　To observe the protective effect of low expression of Gm9866 on high glucose-induced pancreatic β cell injury. Methods　The study was from January to May 2024. Small interfering RNA (siRNA) was used to silence the expression of long-chain non-coding RNA (lncRNA) Gm9866 in MIN6 of mouse pancreatic β cells. The MIN 6 cells were divided into a control group and a high glucose group. The expression of Gm9866 levelin the MIN6 cells was detected by the real-time quantitative polymerase chain reaction (qRT-PCR). The MIN6 cells cultured in high glucose were divided into an si-NC group and an si-Gm9866 group, infected with si-NC lentivirus and si-Gm9866 lentivirus, respectively. Real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression level of Gm9866 in the MIN6 cells. The CCK-8 method and flow cytometry were used to detect the activity and apoptosis of the MIN6 cells. The dual luciferase assay was used to detect the targeting relationship between Gm9866 and miR-149-5p. qRT-PCR was used to detect the expression level of miR-149-5p in the MIN6 cells. Western blot was used to detect the expression levels of apoptotic proteins (BID and BIM) and proliferation proteins (Cdk5 and Cyclin D3). t test was used for the statistical analysis. Results　The expression level of Gm9866 in the control group was lower than that in the high glucose group (1.01±0.29 vs. 7.87±1.20; P<0.01). The MIN6 cells' viability in the si-Gm9866 group was higher than that in the si-NC group (P<0.05); the apoptosis rate in the si-Gm9866 group was lower than that in the si-NC group (P<0.01). Gm9866 targeted and bounded miR-149-5p (P<0.01). The expression level of miR-149-5p in the si-Gm9866 group was higher than that in the si-NC group (P<0.01); the expression levels of apoptotic proteins BID and BIM were lower than those in the si-NC group (both P<0.05); the expression levels of proliferation proteins Cdk5 and Cyclin D3 in the si-Gm9866 group were higher than those in the si-NC group (both P<0.05). Conclusion　Silencing Gm9866 increases the activity of islet cells and reduces the apoptosis of islet cells upregulating the expression of miR-149-5p.

Key words:

Diabetes, LncRNA, Gm9866, MiR-149-5p, Cell proliferation, Cell apoptosis

摘要： 目的　观察低表达Gm9866对高糖诱导的胰岛β细胞损伤的保护作用。方法　研究时 间为2024年1月至5月。以小干扰RNA（siRNA）沉默小鼠胰岛β细胞MIN6细胞中长链非编码RNA （long-chain non-coding RNA，lncRNA）Gm9866的表达。将MIN6细胞分为对照组和高糖组。采用实 时定量聚合酶链式反应（real-time quantitative polymerase chain reaction，qRT-PCR）检测 MIN6 细胞中 Gm9866的表达水平。将高糖培养的MIN6细胞分为si-NC组和si-Gm9866组，分别感染si-NC慢病毒 和si-Gm9866慢病毒。CCK-8法和流式细胞术检测MIN6细胞活性和凋亡情况。双荧光素酶实验检 测 Gm9866 和 miR-149-5p 的 靶 向 关 系 。 qRT-PCR 检 测 MIN6 细 胞 中 miR-149-5p 的 表 达 水 平 。 Western blot检测凋亡蛋白（BID、BIM）和增殖蛋白（Cdk5、Cyclin D3）的表达水平。采用t检验进行统 计分析。结果　对照组MIN6细胞中Gm9866表达水平低于高糖组（1.01±0.29比7.87±1.20，P<0.01）。 si-Gm9866组 MIN6细胞活力高于 si-NC组（P<0.05），细胞凋亡率低于 si-NC组（P<0.01）。Gm9866靶 向结合 miR-149-5p（P<0.01）。si-Gm9866 组 miR-149-5p 表达水平高于 si-NC 组（P<0.01），凋亡蛋白 BID、BIM 表达水平均低于 si-NC 组（均 P<0.05），增殖蛋白 Cdk5、Cyclin D3 表达水平均高于 si-NC 组 （均 P<0.05）。结论　沉默 Gm9866 通过上调 miR-149-5p 表达，增加胰岛细胞活性，减少胰岛细胞 凋亡。

关键词:

糖尿病, lncRNA, Gm9866, miR-149-5p, 细胞增殖, 细胞凋亡