基于自噬探讨龙蛭汤含药血清对H_(2)O_(2)诱导SH-SY5Y神经细胞氧化损伤的保护作用  

Protective effect of Longzhi decoction containing serum on H_(2)O_(2)-induced oxidative damage of SH-SY5Y neurons based on autophagy

作　　者：罗燕萍 陈永斌[1] 刘启华[1] 黄显雯 李南方 范俊逸 张保德 Luo Yanping;Chen Yongbin;Liu Qihua;Huang Xianwen;Li Nanfang;Fan Junyi;Zhang Baode(The First Affiliated Hospital of Guangxi Medical University,Nanning 530021,China)

机构地区：[1]广西医科大学第一附属医院,南宁530021

出　　处：《广西医科大学学报》2021年第5期905-911,共7页Journal of Guangxi Medical University

基　　金：国家自然科学基金资助项目(No.81560790);广西中医药重点学科建设项目(No.GZXK-Z-20-52);陈永斌广西名老中医工作室建设项目(No.桂中医药办发[2019]36号)。

摘　　要：目的:基于自噬探讨龙蛭汤(LZD)含药血清对过氧化氢(H_(2)O_(2))诱导的神经母细胞瘤(SH-SY5Y)氧化损伤的保护作用。方法:采用H_(2)O_(2)诱导SH-SY5Y构建氧化应激损伤模型,观察LZD含药血清对细胞的保护作用。加入自噬激动剂雷帕霉素(RAP)、抑制剂氯喹(CQ)干预SH-SY5Y细胞,观察自噬在SH-SY5Y损伤中的作用,实验分为8组:正常对照组、空白血清(BS)组、模型组、LZD组、RAP组、RAP+LZD组、CQ组、CQ+LZD组。CCK-8法检测各组细胞活力,实时荧光定量PCR(qPCR)法检测自噬相关基因Beclin1、LC3 mRNA相对表达量,Western blotting法比较细胞Beclin1蛋白表达水平及LC3-Ⅱ/LC3-Ⅰ比值。结果:与模型组比较,LZD组细胞存活率显著提高,LZD组Beclin1、LC3 mRNA表达增强,Beclin1蛋白表达及LC3-Ⅱ/LC3-Ⅰ比值升高(均P<0.05)。LZD含药血清协同RAP、CQ干预SH-SY5Y细胞,与CQ组比较,CQ+LZD组Beclin1、LC3 mRNA相对表达量和细胞活力显著升高(均P<0.05);与RAP组比较,RAP+LZD组Beclin1、LC3 mRNA相对表达量降低,细胞活力升高(均P<0.05)。结论:LZD含药血清能够提高神经母细胞瘤细胞活力,并且可能通过调控自噬发挥神经元保护功能。Objective:To observe the protective effect of Longzhidecoction(LZD)-containing serum on oxidative damage induced by hydrogen peroxide(H_(2)O_(2))in neuroblastoma(SH-SY5Y)based on autophagy.Methods:H_(2)O_(2)was used to induce SH-SY5Y to establish an oxidative stress injury model,and the protective effect of LZDcontaining serum on cells was observed.Autophagy agonist Rapamycin(RAP)and inhibitor Chloroquine(CQ)were added to interfere with SH-SY5Y cells,and the role of autophagy in SH-SY5Y injury was observed.The cells were divided into 8 groups:normal control group,blank serum(BS)group,model group,LZD group,RAP group,RAP+LZD group,CQ group,and CQ+LZD group.CCK-8 assay was used to detect the cell viability.qPCR was used to detect the expression levels of autophagy-related genes Beclin1 and LC3.Western blotting was used to determine the protein expressions of Beclin1 and LC3.Results:Compared with model group,the cell survival rate,the mRNA levels of Beclin1 and LC3,the proteinexpression of Beclin1,and the LC3-II/LC3-I ratio were increased in LZD group(P<0.05).The mRNA expression levels of Beclin1 and LC3 and the cell viability in CQ+LZD group were significantly higher than those in CQ group(P<0.05);compared with the RAP group,the expression levels of Beclin1 and LC3 mRNA were decreased while the cell viability was increased in the RAP+LZD group(P<0.05).Conclusion:The serum containing LZD can improve the cell viability of neuroblastoma cells and protect neurons by regulating autophagy.

关 键 词：龙蛭汤含药血清 自噬 神经母细胞瘤细胞 神经元保护 

分 类 号：R963[医药卫生—微生物与生化药学]