眼科新进展2017，Vol.37Issue(9)：805-809,5.DOI:10.13389/j.cnki.rao.2017.0204
补肾活血中药血清对加压纯化培养视网膜神经节细胞PI3K/AKT信号转导通路的影响
Effects of Chinese medicine bushenhuoxue on PI3K/ Akt signal transduction pathway of pressurized and cultured retinal ganglion cells in vitro
摘要
Abstract
Objective To investigate the influence of bushenhuoxue drug-containing serum on PI3K and Akt signaling pathway in purified retinal ganglion cell (RGCs) in vitro of Sprague-Dawley (SD) rats,and to explore the protective mechanisms of bushenhuoxue recipe on RGCs.Methods At first,bushenhuoxue drug-containing serum was prepared,and the RGCs of SD rats were purified;after the apoptotic model of pressurized and purified RGCs was established successfully in vitro using open pressure control system,RGCs were dealt with 50 g · L-1,100 g · L-1,200 g · L-1 concentration gradient of bushenhuoxue drug-containing serum.Then the subjected cells were divided into normal culture group (N group),control group (C group),50 g · L-1 bushenhuoxue group (50 g · L-1 BSHX group),100 g · L-1 bushenhuoxue group (100 g · L-1 BSHX group),200 g · L-1 bushenhuoxue group (200 g · L-1 BSHX group).Finally,cell apoptotic rate was detected by Annexin V-FITC/PI staining,while real-time quantitative PCR (qRT-PCR) and Western blot were used to detect the mRNA and protein expression of PI3K and Akt in each group respectively.Results The results of qRT-PCR detection showed that PI3K,Akt mRNA expression level in C group (0.04 ±0.01) was decreased compared with N group (1.00 ± 0.04),and the difference was statistically significant (all P＜0.05),while PI3K,Akt mRNA levels in 50 g · L-1,100 g · L-1 and 200 g · L-1 BXHX group (0.18 ±0.01,0.21 ±0.02,0.22 ±0.01,0.36 ±0.01,0.84 ±0.10,1.07 ± 0.17) were increased compared with the C group,and the difference was statistically significant (all P ＜0.05).The Western blot results of each group showed that PI3K,Akt protein expression level in C group was decreased compared with N group,with statistical difference (all P ＜ 0.05),while PI3 K,Akt protein expression levels in 50 g · L-1,100 g · L-1 and 200 g · L-1 BSHX group were increased compared with C group,with staffstical difference (all P ＜ 0.05).Conclusion Bushenhuoxue drug-containing serum may inhibit the RGCs apoptosis induced by pressure,which may be related to the activation of PBK/Akt signal transduction pathway.关键词
视网膜神经节细胞/中药/补肾活血/PI3K/Akt/加压纯化培养/信号转导通路Key words
retinal ganglion cells/traditional Chinese medicine/bushenhuoxue/PI3K/Akt/pressurized and purified culture/signal transduction pathway分类
医药卫生引用本文复制引用
汪伟,李翔,刘红佶,牟琳,张丹..补肾活血中药血清对加压纯化培养视网膜神经节细胞PI3K/AKT信号转导通路的影响[J].眼科新进展,2017,37(9):805-809,5.基金项目
国家自然科学基金资助(编号:81373695)National Natural Science Foundation of China (No:81373695) （编号:81373695）
