基于PI3K/AKT通路探讨青光增视方对过氧化氢诱导的视网膜神经节细胞损伤的保护机制  

Protective effect of Qingguang Zengshi prescription on hydrogen peroxide-induced injuries of retinal ganglion cell in vitro based on PI3K/AKT pathway

作　　者：姜艳华 赵磊[1,3] 陈琳琳[2] 左韬[1,3] JIANG Yanhua;ZHAO Lei;CHEN Linlin;ZUO Tao(Department of Ophthalmology,The Second Clinical College of Liaoning University of Traditional Chinese Medicine,Shenyang 110032,China;Department of Ophthalmology,The Fourth People’s Hospital of Shenyang,China Medical University,Shenyang 110031,China;Department of Ophthalmology,The Second Affiliated Hospital of Liaoning University of Traditional Chinese Medicine,Shenyang 110034,China)

机构地区：[1]辽宁中医药大学第二临床学院眼科,沈阳110032 [2]中国医科大学沈阳市第四人民医院眼科,沈阳110031 [3]辽宁中医药大学附属第二医院眼科,沈阳110034

出　　处：《中国医科大学学报》2023年第8期706-710,717,共6页Journal of China Medical University

基　　金：辽宁省自然科学基金(2018010548-301)。

摘　　要：目的探讨青光增视方对过氧化氢(H2O2)诱导的视网膜神经节细胞(RGC)-5损伤的保护作用及其对磷脂酰肌醇3激酶/蛋白激酶B(PI3K/AKT)通路的调节作用。方法通过H2O2诱导体外培养的RGC-5建立氧化应激损伤细胞模型,用空白血清(模型组)、甲钴胺含药血清(西药组)、青光增视方高、中、低剂量含药血清(青光增视方组)处理模型细胞。采用CCK-8法检测RGC-5的存活率,流式细胞术检测RGC-5的凋亡情况,采用定量PCR及Western blotting检测RGC-5的PI3K、AKT、PTEN、eNOS mRNA及蛋白的表达情况。结果与空白组比较,各组RGC-5存活率降低,模型组PI3K、eNOS mRNA与PI3K、p-AKT/AKT、eNOS蛋白表达水平降低,PTEN mRNA及蛋白表达升高;与模型组比较,西药组、青光增视方高、中剂量组RGC-5存活率增加,西药组、青光增视方高剂量组PI3K、eNOS mRNA及中剂量组的eNOS mRNA表达升高,各给药组PTEN mRNA及蛋白表达降低,PI3K、p-AKT/AKT、eNOS蛋白表达升高;与西药组比较,青光增视方低、中剂量组PTEN mRNA表达升高,eNOS mRNA降低(P<0.05),高剂量组PI3K、p-AKT/AKT及中剂量组p-AKT/AKT蛋白升高,青光增视方高、中剂量组PTEN蛋白降低,差异均有统计学意义(均P<0.05)。结论青光增视方对H2O2诱导的RGC损伤具有保护作用,其机制可能与调控PI3K、AKT、PTEN、eNOS的表达有关。Objective To analyze the effect of Qingguang Zengshi prescription on hydrogen peroxide-induced injuries to retinal ganglion cells(RGCs)in vitro and on the phosphatidylinositol 3 kinase(PI3K)/protein kinase B(AKT)signal pathway and its related factors.Methods Hydrogen peroxide was applied to RGC-5 to create an oxidative damage cell model.Blank serum(model group),mecobalamin-medicated serum(western medicine group),and low-,medium-,and high-dose Qingguang Zengshi prescription-medicated serum(Qingguang Zengshi administration group)were applied to the oxidative damage cell model(RGC-5).CCK-8 was used to detect the survival rate;flow cytometry detected the apoptosis of RGC-5;and qRT-PCR and Western blotting were used to detect PI3K,AKT,PTEN,and eNOS mRNA and protein expressions.Results Compared with those in the blank group,RGC-5 survival rates in all other groups were lower(P<0.05);PI3K and eNOS mRNA and PI3K,p-AKT/AKT,and eNOS protein expression levels in the model group were lower(P<0.05);and PTEN mRNA and protein expression levels in the model group were higher(P<0.05).Compared with those in the model group,the survival rates of RGC-5 in the mecobalamin group and the high-and middle-dosage Qingguang Zengshi prescription groups were higher(P<0.05);the expression levels of PI3K and eNOS mRNA in the western-medicine group and the high-dosage Qingguang Zengshi prescription group and eNOS mRNA in the middle-dosage group were higher(P<0.05);the expression levels of PTEN mRNA and protein in each administration group were lower(P<0.05);and the expression levels of PI3K,p-AKT/AKT and eNOS protein in each administration group were higher(P<0.05).Compared with those in the mecobalamin group,the PTEN mRNA expression levels in the low-and middle-dosage Qingguang Zengshi prescription groups were higher(P<0.05);eNOS mRNA expression level was higher(P<0.05);PI3K and p-AKT/AKT in the high-dosage group and p-AKT/AKT in the middle-dosage group were higher(P<0.05);and PTEN protein levels in the high-and middle-dosage groups were lower

关 键 词：青光眼 视网膜神经节细胞 青光增视方 磷脂酰肌醇3激酶 蛋白激酶B 

分 类 号：R243[医药卫生—中医临床基础]