枸杞多糖通过增强自噬抑制饥饿诱导的施万细胞凋亡
Lycium barbarum polysaccharide inhibits starvation-induced apoptosis of Schwann cells by enhancing autophagy
刘玉梅 1王贺颖 1魏财鸿 1王进娟 1李军平 1贾桦1
作者信息
- 1. 宁夏医科大学基础医学院人体解剖与组织胚胎学系,银川 750004
- 折叠
摘要
目的:探讨枸杞多糖(LBP)抑制施万细胞(SCs)凋亡的作用及相关机制.方法:RSC96细胞饥饿处理12 h制备自噬模型,Western Blot检测自噬相关蛋白LC3、p62的表达;采用Cell Counting Kit-8(CCK-8)试剂盒筛选出LBP作用的最佳浓度.将RSC96细胞随机分为正常组(Control)、饥饿组(Starvation)、Starvation+LBP组,Western Blot或免疫荧光染色检测细胞自噬相关蛋白(LC3、p62)以及髓鞘相关蛋白(p75NTR、PMP22、S100β)的表达;采用Annexin V/PI荧光染色检测细胞的凋亡情况;应用流式细胞术分析细胞周期;Western Blot分析通路蛋白Erk1/2、Akt的磷酸化水平.结果:CCK8结果显示,LBP为300 μg/ml时受损RSC96细胞活力最佳.与 Control 组相比,Starvation 组 LC3-Ⅱ/LC3-I 水平显著增高(P＜0.05).与 Starvation 组相比,Starvation+LBP组的凋亡和坏死细胞比例显著降低,处于S期和G2/M期的细胞比例升高;LC3-Ⅱ、p75NTR、PMP22、S100β蛋白表达量增高,自噬底物蛋白p62的表达量降低;通路蛋白p-Erk1/2表达量增高(P＜0.05),而p-Akt蛋白表达量略有下降.结论:LBP可通过增强细胞自噬抑制SCs凋亡,促进髓鞘相关蛋白的表达,其机制与Erk1/2激活和/或Akt抑制有关.
Abstract
Objective:To investigate the inhibitory effect of lycium barbarum polysaccharide(LBP)on apoptosis of Schwann cells(SCs)and its related mechanisms.Methods:The autophagy model was prepared by starvation treatment of RSC96 cells for 12 h,and the expressions of autophagy related proteins LC3 and p62 were detected by Western Blot.Cell Counting Kit-8(CCK-8)kits were used to detect the optimal concentration of LBP.RSC96 cells were randomly divided into Control group,Starvation group and Starvation+LBP group.The expressions of autophagy associated pro-teins(LC3,p62)and myelin associated proteins(p75NTR,PMP22,S100β)were detected by Western Blot or immu-nofluorescence staining.Annexin V/PI fluorescence staining was used to detect apoptosis of the cells.The cell cycle was analyzed by flow cytometry.Western Blot analysis of phosphorylation levels of pathway proteins Erk1/2 and Akt.Results:CCK8 results showed that the viability of damaged RSC96 cells was the best when LBP was 300 μg/ml.Com-pared with Control group,LC3-Ⅱ/LC3-I levels in Starvation group were significantly increased(P＜0.05).Compared with Starvation group,the proportion of apoptotic and necrotic cells in Starvation+LBP group was significantly de-creased,and the proportion of cells in S and G2/M stages was increased.The expression levels of LC3-Ⅱ,p75NTR,PMP22 and S100β were increased,while the expression levels of autophagy substrate protein p62 were decreased.In-creased expression of pathway protein p-Erk1/2(P＜0.05),while the expression of p-Akt protein decreased slightly.Conclusion:LBP can inhibit the apoptosis of SCs and promote the expression of myelin-related proteins by enhancing autophagy,which is related to the activation of Erk1/2 and/or the inhibition of Akt.
关键词
枸杞多糖/施万细胞/自噬/凋亡/大鼠Key words
lycium barbarum polysaccharide(LBP)/Schwann cells(SCs)/autophagy/apoptosis/rat引用本文复制引用
出版年
2024
NETL
NSTL
万方数据