摘要
目的:研究三七总皂苷(PNS)抗氧化应激诱导的血管内皮细胞自噬作用机制。方法:将人脐静脉内皮细胞(HUVECs)分为空白组、模型组和治疗组,运用qRT-PCR和Western blot检测自噬蛋白6(Beclin1)、自噬蛋白5(Atg5)mRNA和蛋白表达及细胞自噬流情况;利用透射电子显微镜观察细胞自噬小体形成情况。结果:与空白组比较,模型组能促进自噬小体形成,增加自噬相关膜的合成,上调Beclin1 mRNA和蛋白表达(P<0.05,P<0.01),同时提高Atg5蛋白水平(P<0.01),下调Atg5 mRNA表达(P<0.01)。与模型组比较,治疗组能抑制自噬小体形成,减少细胞自噬流和自噬相关膜合成,并显著下调Beclin1 mRNA和蛋白表达水平(P<0.05),上调Atg5 mRNA表达(P<0.01)。结论:PNS可以通过抑制Beclin1 mRNA及蛋白表达水平,抑制细胞自噬小体和自噬流,从而发挥抗H2O2诱导的HUVECs细胞自噬作用。
Objective:To study the mechanism of anti-oxidative stress-induced autophagy in human umbilical vein endothelial cells(HUVECs)by panax notoginseng saponins(PNS).Methods:The experiment was divided into control group,model group and treatment group.The mRNA and protein expressions of Beclinl and Atg5 as well as the autophagy flow were detected by qRT-PCR and Western blot,respectively.Transmission electron microscope was applied to observe the formation of autophagosomes.Results:Compared with the control group,the model group could promote the formation of autophagosomes and the synthesis of autophagy-related membranes,the model group also up-regulated the expressions of Beclinl mRNA and protein(P<0.05,P<0.01)as well as Atg5 protein(P<0.01),but down-regulated the expression of Atg5 mRNA(P<0.01).Compared with the model group,the PNS treatment group inhibited the formation of autophagosomes,the activation of cell autophagy flux and the synthesis of autophagy-related membrane.Moreover,PNS treatment group could significantly down-regulate the expressions of Beclinl mRNA and protein(P<0.05),but up-regulate Atg5 mRNA expression(P<0.01).Conclusion:PNS could inhibit autophagosomes and autophagic flux by inhibiting the mRNA and protein expression levels of Beclin1,thereby resisting H_(2)O_(2)-induced autophagy of HUVECs.
作者
董艳
李军
张振鹏
刘咏梅
王阶
DONG Yan;LI Jun;ZHANG Zhenpeng;LIU Yongmei;WANG Jie(Guang'anmen Hospital,China Academy of Chinese Medical Sciences,Beijing 100053,China)
出处
《中华中医药杂志》
CAS
CSCD
北大核心
2023年第10期4683-4687,共5页
China Journal of Traditional Chinese Medicine and Pharmacy
基金
中国中医科学院优秀青年科技人才(创新类)培养专项(No.ZZ14-YQ-018)。
