摘要
目的:探讨当归多糖对高糖诱导的视网膜神经节细胞凋亡的抑制作用及其可能机制。方法:采用不同浓度当归多糖干预视网膜神经节细胞RGC-5,MTT法检测细胞存活情况并筛选合适浓度。培养RGC-5细胞并添加50 mmol/L葡萄糖建立高糖损伤模型后,分为高糖组、当归多糖组和当归多糖+Nrf2信号通路抑制剂(ML385)组,另设对照组。当归多糖组使用100μmol/L的当归多糖处理,当归多糖+ML385组使用100μmol/L的当归多糖联合20μmol/L的ML385处理,高糖组使用50 mmol/L葡萄糖处理,对照组加入等量完全培养基。CCK-8法检测细胞存活情况;流式细胞术检测细胞凋亡率;ELISA法检测各组细胞中丙二醛(MDA)、谷胱甘肽过氧化物酶(GSH-Px)、超氧化物岐化酶(SOD)含量;实时定量PCR法检测细胞中蛋白激酶B(Akt)mRNA、糖原合成酶激酶-3β(GSK-3β)mRNA和核因子E2相关因子2(Nrf2)mRNA表达情况;Western blotting检测凋亡蛋白B细胞淋巴瘤因子2(Bcl-2)、Bcl-2相关蛋白(Bax)、Caspase-3和Caspase-9蛋白表达情况。结果:当归多糖组细胞活力、GSH-Px含量、SOD含量、Nrf2 mRNA相对表达量、Bcl-2蛋白相对表达量均高于高糖组,细胞凋亡率、MDA含量、Akt mRNA相对表达量、GSK-3βmRNA相对表达量、Bax蛋白相对表达量、Caspase-3蛋白相对表达量、Caspase-9蛋白相对表达量均低于高糖组(P<0.05)。当归多糖+ML385组细胞活力、GSH-Px含量、SOD含量、Nrf2 mRNA相对表达量、Bcl-2蛋白相对表达量均低于当归多糖组,细胞凋亡率、MDA含量、Akt mRNA相对表达量、GSK-3βmRNA相对表达量、Bax蛋白相对表达量、Caspase-3蛋白相对表达量、Caspase-9蛋白相对表达量均高于当归多糖组(P<0.05)。结论:当归多糖能抑制高糖诱导的RGC-5细胞凋亡,降低细胞内氧化应激水平,可能机制为通过阻滞Akt/GSK-3β通路,提高细胞内抗氧化Nrf2表达水平。
Objective:To investigate the inhibitory effect of angelica polysaccharide on retinal ganglion cell apoptosis induced by high glucose and its possible mechanism.Methods:RGC-5 cells were treated with different concentrations of angelica polysaccharide.The survival of RGC-5 cells was detected by MTT assays to identify the appropriate concentration of angelica polysaccharide.RGC-5 cells were cultured and treated with 50 mmol/L glucose to establish a high glucose model.They were divided into high glucose group,angelica polysaccharide group,angelica polysaccharide+Nrf2 signal pathway inhibitor(ML385)group.Set up another control group.The angelica polysaccharide group was treated with 100μmol/L angelica polysaccharide,the angelica polysaccharide+ML385 group was treated with 100μmol/L angelica polysaccharide combined with 20μmol/L ML385,the high glucose group was treated with 50 mmol/L glucose,and the control group was treated with equal amounts of complete medium.Cell survival was detected by CCK-8 assays.The apoptosis rate was detected by flow cytometry.The levels of malondialdehyde(MDA),glutathione peroxidase(GSH-Px)and superoxide dismutase(SOD)were detected by ELISA.The mRNA expressions of protein kinase B(Akt),glycogen synthase kinase-3β(GSK-3β)and nuclear factor E2 related factor 2(Nrf2)were detected by real-time quantitative PCR.The expressions of apoptotic protein B-cell lymphoma factor 2(Bcl-2),Bcl-2 related protein(Bax),Caspase-3 and Caspase-9 were detected by Western blotting.Results:The cell viability,GSH-Px and SOD content,levels of Nrf2 mRNA and Bcl-2 protein were higher in the angelica polysaccharide group than those in the high glucose group,and the apoptosis rate,MDA content,levels of Akt mRNA,GSK-3βmRNA,Bax protein,Caspase-3 protein and Caspase-9 protein were lower than those in the high glucose group(P<0.05).The cell viability,GSH-Px and SOD content,levels of Nrf2 mRNA and Bcl-2 protein were lower in the angelica polysaccharide+ML385 group than in the angelica polysaccharide group,and the apoptos
作者
王艳新
贾冠美
曹朗
曹顺义
张士宏
WANG Yanxin;JIA Guanmei;CAO Lang;CAO Shunyi;ZHANG Shihong(Baoding Second Central Hospital,Baoding Hebei 072750,China)
出处
《中医药导报》
2023年第5期16-21,共6页
Guiding Journal of Traditional Chinese Medicine and Pharmacy
基金
保定市科技计划项目(2141ZF218)。
