Li R-X, Wang Q, Shen Z-W, Jie L-M
Department of Ophometry, Xiamen University Affiliated Xiamen Eye Center, Xiamen, Fujian, China.
Eur Rev Med Pharmacol Sci. 2020 May;24(10):5209-5216. doi: 10.26355/eurrev_202005_21302.
Retinal ganglion cells (RGCs) are the main cells that form vision in the retina. MT1M is involved in the occurrence and progression of various diseases. However, the role of MTIM in RGCs cells remains unclear.
RGCs were cultured in vitro and randomly divided into control group, MT1M group (transfected with MT1M-pcDNA3.1 plasmid), and MT1M siRNA group (transfected with MT1M siRNA) followed by measuring MT1M and NT-3 expression by real time PCR and Western blot, cell proliferation by MTT assay, secretion of IL-2 and IL-6 by enzyme-linked immunosorbent assay (ELISA), SOD activity and ROS content. In addition, expression of PI3K/AKT signaling pathway protein was detected by Western blot.
MT1M expression in MT1M group was significantly increased, which promoted cell proliferation, increased NT-3 expression, and decreased Caspase 3 activity and IL-2 and IL-6 secretion. Meanwhile, SOD activity was increased, ROS content was decreased and PI3K/AKT protein phosphorylation was elevated. The differences were statistically significant compared with control group (p < 0.05). MT1M siRNA decreased MT1M expression, inhibited cell proliferation, decreased NT-3, and increased Caspase 3 activity and IL-2 and IL-6 secretion. In addition, MT1M siRNA decreased SOD activity, increased ROS content and reduced PI3K/AKT protein phosphorylation. Compared with control group, the differences were statistically significant (p < 0.05).
Up-regulation of MT1M can inhibit RGC cell apoptosis and inflammation, and promote RGC cell proliferation through the PI3K/AKT signaling pathway.
视网膜神经节细胞(RGCs)是视网膜中形成视觉的主要细胞。MT1M参与多种疾病的发生和发展。然而,MTIM在RGCs细胞中的作用仍不清楚。
体外培养RGCs,随机分为对照组、MT1M组(转染MT1M-pcDNA3.1质粒)和MT1M siRNA组(转染MT1M siRNA),然后通过实时PCR和蛋白质印迹法检测MT1M和NT-3表达,通过MTT法检测细胞增殖,通过酶联免疫吸附测定(ELISA)检测IL-2和IL-6分泌、SOD活性和ROS含量。此外,通过蛋白质印迹法检测PI3K/AKT信号通路蛋白的表达。
MT1M组中MT1M表达显著增加,促进细胞增殖,增加NT-3表达,并降低Caspase 3活性以及IL-2和IL-6分泌。同时,SOD活性增加,ROS含量降低,PI3K/AKT蛋白磷酸化升高。与对照组相比,差异具有统计学意义(p<0.05)。MT1M siRNA降低MT1M表达,抑制细胞增殖,降低NT-3,并增加Caspase 3活性以及IL-2和IL-6分泌。此外,MT1M siRNA降低SOD活性,增加ROS含量,并降低PI3K/AKT蛋白磷酸化。与对照组相比,差异具有统计学意义(p<0.05)。
MT1M的上调可抑制RGC细胞凋亡和炎症,并通过PI3K/AKT信号通路促进RGC细胞增殖。