Department of Ophthalmology, Second Hospital of Jilin University, Changchun, 130041, China.
Department of Pharmacology, College of Basic Medical Sciences, Jilin University, Changchun 130021, China.
Biomed Pharmacother. 2018 Dec;108:1201-1207. doi: 10.1016/j.biopha.2018.09.140. Epub 2018 Oct 2.
Berberine (BBR) has beneficial effects on diabetes and the multiple complications of diabetes due to its anti-apoptotic activity; however, the effect of BBR on diabetic retinopathy and its mechanism of action have not been clarified. The present study investigated the effect of BBR on Müller cells stimulated with high glucose (HG). Primary retinal Müller cells were incubated with high glucose to induce cell apoptosis; cells were pretreated with the AMPK inhibitor compound C and the AMPK activator AICAR to further explore the role of the AMPK/mTOR signaling pathway in the anti-apoptotic action of BBR. Immunofluorescence was used to measure apoptosis and autophagy. Western blot analysis was employed to determine the levels of p-AMPK and p-mTOR, as well as apoptosis-related proteins and autophagy-related proteins in Müller cells. Our results showed that BBR attenuated apoptosis, up regulated Bcl-2 and down regulated Bax and caspase-3 expression; enhanced the formation of autophagy, elevated the expression of Beclin-1 and LC3II and activated the AMPK/mTOR signaling pathway in Müller cells under high glucose conditions compared with the control group. The effect of BBR was partly blocked by compound C and strengthened by AICAR. BBR may have therapeutic potential to protect Müller cells from high-glucose-inducing apoptosis through enhancing autophagy and activating the AMPK/mTOR signaling pathway.
小檗碱(BBR)具有抗细胞凋亡活性,对糖尿病及其多种并发症有有益作用;然而,BBR 对糖尿病性视网膜病变的作用及其作用机制尚未阐明。本研究探讨了 BBR 对高糖(HG)刺激的 Muller 细胞的作用。原代视网膜 Muller 细胞在高糖中孵育以诱导细胞凋亡;用 AMPK 抑制剂化合物 C 和 AMPK 激活剂 AICAR 预处理细胞,进一步探讨 AMPK/mTOR 信号通路在 BBR 抗细胞凋亡作用中的作用。免疫荧光法用于测量细胞凋亡和自噬。Western blot 分析用于测定 Muller 细胞中 p-AMPK 和 p-mTOR 以及凋亡相关蛋白和自噬相关蛋白的水平。结果表明,BBR 可减轻细胞凋亡,上调 Bcl-2 并下调 Bax 和 caspase-3 表达;与对照组相比,BBR 可增强高糖条件下 Muller 细胞的自噬形成,上调 Beclin-1 和 LC3II 的表达,并激活 AMPK/mTOR 信号通路。BBR 的作用部分被化合物 C 阻断,AICAR 增强。BBR 可能通过增强自噬并激活 AMPK/mTOR 信号通路,具有保护 Muller 细胞免受高糖诱导的细胞凋亡的治疗潜力。