a Department of Nephrology, Jining No.1 People's Hospital , Jining , China.
b Affiliated Jining No.1 People's Hospital of Jining Medical University, Jining Medical University , Jining , China.
Artif Cells Nanomed Biotechnol. 2019 Dec;47(1):2940-2947. doi: 10.1080/21691401.2019.1640231.
Arbutin (ARB) has been widely used in skin pigmentation disorders. Nevertheless, the involvements of ARB in diabetic nephropathy (DN) are still unknown. We investigated the functions of ARB in high glucose (HG)-induced cell apoptosis and autophagy in HK-2 cells. Cell viability was examined through CCK-8 in HK-2 cells after disposal with 45 mM glucose and ARB (10-50 μM). Flow cytometry and western blot tested cell apoptosis and the related protein levels in HK-2 cells after 45 mM glucose and 50 μM ARB administration. RT-qPCR delved microRNA (miR)-27a expression in HG and ARB co-treated HK-2 cells. Effect of miR-27a on ARB affected cell apoptosis and autophagy was investigated after miR-27a inhibitor transfection. JNK and mTOR pathways were finally assessed by western blot. ARB alleviated HG-induced cell apoptosis, autophagy and regulated the related protein levels in HK-2 cells. MiR-27a expression was reduced in HG-treated cells, but was accelerated in HG and ARB co-treated HK-2 cells with the increased concentration. Inhibition of miR-27a apparently abolished the outcomes of ARB in HG-induced HK-2 cells apoptosis and autophagy. Besides, ARB blocked JNK and mTOR pathways by regulating miR-27a. The findings demonstrated that ARB alleviated apoptosis and autophagy in HG-treated HK-2 cells by regulating miR-27a/JNK/mTOR axis.
熊果苷 (ARB) 已广泛用于皮肤色素沉着紊乱。然而，ARB 在糖尿病肾病 (DN) 中的作用仍不清楚。我们研究了 ARB 在高葡萄糖 (HG) 诱导的 HK-2 细胞凋亡和自噬中的作用。用 45mmol/L 葡萄糖和 ARB（10-50μM）处理 HK-2 细胞后，通过 CCK-8 检测细胞活力。流式细胞术和 Western blot 检测 45mmol/L 葡萄糖和 50μM ARB 处理后 HK-2 细胞的细胞凋亡和相关蛋白水平。RT-qPCR 检测 HG 和 ARB 共同处理的 HK-2 细胞中 microRNA (miR)-27a 的表达。转染 miR-27a 抑制剂后，研究 miR-27a 对 ARB 影响细胞凋亡和自噬的作用。最后通过 Western blot 评估 JNK 和 mTOR 通路。ARB 减轻了 HG 诱导的 HK-2 细胞凋亡、自噬，并调节了 HK-2 细胞中相关蛋白的水平。miR-27a 在 HG 处理的细胞中表达减少，但在 HG 和 ARB 共同处理的 HK-2 细胞中表达增加。miR-27a 抑制剂的抑制作用明显消除了 ARB 在 HG 诱导的 HK-2 细胞凋亡和自噬中的作用。此外，ARB 通过调节 miR-27a/JNK/mTOR 轴来阻断 JNK 和 mTOR 通路。研究结果表明，ARB 通过调节 miR-27a/JNK/mTOR 轴减轻 HG 处理的 HK-2 细胞中的凋亡和自噬。
