主 办:北 京 中 医 药 大 学
ISSN 1006-2157 CN 11-3574/R

北京中医药大学学报 ›› 2020, Vol. 43 ›› Issue (8): 661-667.doi: 10.3969/j.issn.1006-2157.2020.08.008

• 中药药理 • 上一篇    下一篇

益气养血疏肝方对谷氨酸钠损伤的视网膜神经节细胞的保护作用*

李甜甜1, 周剑2#, 闫晓玲2, 苏艳2, 周晓昱1, 韦企平2   

  1. 1 北京中医药大学 北京 100029;
    2 北京中医药大学东方医院眼科
  • 收稿日期:2020-04-21 出版日期:2020-08-30 发布日期:2020-08-27
  • 通讯作者: # 周剑,男,硕士,教授,主任医师,博士生导师,主要研究方向:中西医结合治疗视神经病变,E-mail:zhj9667@163.com
  • 作者简介:李甜甜,女,在读博士生
  • 基金资助:
    * 国家自然科学基金面上项目(No.81874491)

Effect of Yiqi Yangxue Shugan Formula on glutamate-induced RGC-5 cell injury model *

Li Tiantian1, Zhou Jian2#, Yan Xiaoling2, Su Yan2, Zhou Xiaoyu1, Wei Qiping2   

  1. 1 Beijing University of Chinese Medicine, Beijing 100029, China;
    2 Department of Ophthalmology, Dongfang Hospital, the Second Affiliated Hospital of Beijing University of Chinese Medicine, Beijing 100078, China
  • Received:2020-04-21 Online:2020-08-30 Published:2020-08-27
  • Contact: Prof. Zhou Jian, M.D., Chief Physician of Ophthalmology Department, Dongfang Hospital, the Second Affiliated Hospital of Beijing University of Chinese Medicine, Beijing 100078. E-mail:zhj9667@126.com
  • Supported by:
    National Natural Science Foundation of China (No. 81874491)

摘要: 目的 研究益气养血疏肝方(青盲一号方)抑制谷氨酸钠诱导的视网膜神经节细胞(RGC-5)凋亡的作用及可能机制。方法 星孢菌素(STSN)分化体外培养的RGC-5细胞后,L-谷氨酸钠诱导RGC-5细胞损伤模型,分别用2.5%、5%、10%益气养血疏肝方药物血清干预24 h。采用CCK-8检测各组细胞活性;流式细胞术检测各组细胞凋亡率;Western-blot检测Bcl-2/Bax信号通路相关蛋白的表达水平;RT-PCR检测Bcl-2、Bax基因表达。结果 各药物血清组的细胞存活率均高于模型组(P<0.05),且5%益气养血疏肝方药物血清组的存活率高于其他药物血清组(P<0.05),同时凋亡率最低。与空白对照组比较,模型组Bcl-2蛋白表达下降(P<0.05),Caspase-9、Caspase-3蛋白表达增加(P<0.05)。与模型组比较,5%益气养血疏肝方药物血清组Bcl-2蛋白表达增加,Caspase-9、Caspase-3蛋白表达下降(P<0.05),且5%药物血清组Bcl-2/Bax高于2.5%、10%药物血清组。结论 5%益气养血疏肝方药物血清能够通过调控Bcl-2/Bax信号通路来抑制RGC-5细胞的凋亡。

关键词: 益气养血疏肝方, 谷氨酸, 视网膜神经节细胞, 凋亡, Bcl-2/Bax信号通路

Abstract: Objective To evaluate the effect of Yiqi Yangxue Shugan Formula (also known as Qing Mang Yi Hao Formula, QMYH) on inhibiting apoptosis of RGC-5 cells by detecting the apoptosis rate in RGC-5 glutamate-induced injury model and related mechanism. Methods Staurosporine (STSN) was used to differentiate RGC-5 cells in vitro. Sodium L-glutamate was used to establish RGC-5 cell apoptosis model, and the cell culture medium containing QMYH serum was used for intervention. CCK-8 Kit was used to detect the survival rate of RGC-5 cells in each group; flow cytometry was used to detect the apoptosis rate of cells in each group; Western-blot was used to analyze the expression levels of Bcl-2/Bax signaling pathway-related proteins in RGC-5 cells; RT-PCR detected the mRNA expression levels of Bcl-2 and Bax. Results Intervention with 0.125 μmol/L STSN medium for 1 h could induce RGC-5 neuronal differentiation. 8 mmol/L sodium L-glutamate interfering for 18 h could cause 50% of RGC-5 cells injury. The medium-dose QMYH group (5% QMYH group) had the highest survival rate of RGC-5 cells (51.1%) and the lowest apoptosis rate (3.9%); compared with the low-dose and high-dose QMYH groups, the mRNA expression level of Bcl-2 in the middle-dose group was obviously up-regulated (P> 0.05); in addition, the protein Bcl-2/Bax ratio increased significantly. Conclusion The 5% QMYH serum could possibly inhibit the apoptosis of RGC-5 cells by regulating the Bcl-2/Bax signaling pathway.

Key words: Yiqi Yangxue Shugan Formula (Qing Mang Yi Hao Formula, QMYH), glutamate, RGC-5 cells, apoptosis, Bcl-2/Bax signaling pathway

中图分类号: 

  • R285.5