摘要
目的探索人参皂苷Rk3在减轻视网膜色素上皮(retinal pigment epithelial,RPE)细胞衰老及治疗新生血管性老年性黄斑变性(neovascular age-related macular degeneration,nAMD)的作用及机制。方法通过过氧化氢诱导ARPE-19细胞衰老模型和nAMD小鼠模型,验证人参皂苷Rk3减轻RPE细胞衰老和缩小脉络膜新生血管(choroidal neovascularization,CNV)面积的药物活性;通过转录组测序分析差异基因表达谱变化;通过蛋白免疫印迹实验检测mTOR、S6K1、p-S6K1、4EBP1、p-4EBP1、p62、LC3A/B等蛋白的表达水平;通过分子对接和动力学模拟分析人参皂苷Rk3与mTOR蛋白的亲和力。结果①细胞实验发现人参皂苷Rk3显著减轻ARPE-19细胞衰老(P<0.05);②动物实验证实人参皂苷Rk3显著缩小nAMD小鼠视网膜CNV面积(P<0.01),且与临床一线治疗药物阿柏西普效果相当(P>0.05)。③转录组测序提示人参皂苷Rk3导致PI3K-Akt信号通路富集,蛋白免疫印迹实验显示人参皂苷Rk3可抑制mTORC1信号的过度激活(P<0.05),同时增强细胞自噬(P<0.05)。④分子对接和动力学模拟研究进一步验证人参皂苷Rk3很可能靶向mTOR蛋白减轻RPE细胞衰老。结论人参皂苷Rk3通过抑制mTOR增强细胞自噬,减轻了RPE细胞衰老并缩小了nAMD小鼠视网膜CNV面积。
Objective To investigate the role and mechanism of ginsenoside Rk3 in alleviating the senescence of retinal pigment epithelial(RPE)cells and in treating neovascular age-related macular degeneration(nAMD).Methods An ARPE-19 cell senescence model induced by hydrogen peroxide and an nAMD mouse model were subjected to validate the pharmacological activity of ginsenoside Rk3 in reducing RPE cell senescence and shrinking choroidal neovascularization(CNV)area.Transcriptome sequencing was performed to analyze the changes in differential gene expression profiles.Western blotting was used to detect the expression levels of mTOR,S6K1,p-S6K1,4EBP1,p-4EBP1,p62,and LC3A/B.Molecular docking and dynamics simulations were conducted to analyze the binding affinity of ginsenoside Rk3 with mTOR protein.Results①Cell experiments showed that ginsenoside Rk3 significantly alleviated the senescence in ARPE-19 cells(P<0.05).②Animal experiments confirmed that ginsenoside Rk3 significantly reduced the retinal CNV area in nAMD mice(P<0.01),and its efficacy was comparable to the first-line clinical drug aflibercept(P>0.05).③Transcriptome sequencing suggested that ginsenoside Rk3 led to enrichment of the PI3K-Akt signaling pathway.Western blotting showed that ginsenoside Rk3 inhibited the overactivation of mTORC1 signaling(P<0.05)and simultaneously enhanced cellular autophagy(P<0.05).④Molecular docking and dynamics simulations further validated that ginsenoside Rk3 probably targets mTOR to alleviate RPE cell senescence.Conclusion Ginsenoside Rk3 alleviates RPE cell senescence and reduces retinal CNV area in nAMD mice by enhancing autophagy through inhibition of mTOR.
作者
赫飞翔
刘雪梅
唐倩
刘莛
HE Feixiang;LIU Xuemei;TANG Qian;LIU Ting(Department of Ophthalmology,Army Medical Center of PLA/Daping Hospital of Third Military Medical University,Chongqing;Department of Ophthalmology,No.904 Hospital of Joint Logistics Support Force,Wuxi,Jiangsu;School of Pharmacy and Bioengineering,Chongqing University of Technology,Chongqing,China)
出处
《陆军军医大学学报》
2025年第19期2340-2350,共11页
Journal of Army Medical University
基金
重庆市自然科学基金面上项目(CSTB2024NSCQ-KJFZZDX0038)。
作者简介
通信作者:刘莛,Email:liuting0727@tmmu.edu.cn。
