活血散结方对PDGF干预下兔RPE细胞凋亡及caspase-3、P53表达的影响  

Effect of Huoxue Sanjie Recipe on Apoptosis and Expression of Caspase-3 and p53 in Rabbit RPE Cells Under PDGF Intervention

作　　者：刘晓清 李建超 谭涵宇[1] 彭俊[4] 彭清华[1,2] 吴权龙 LIU Xiao-qing;LI Jian-chao;TAN Han-yu;PENG Jun;PENG Qing-hua;WU Quan-long(Hunan University of Chinese Medicine,Changsha 410000,China;Hunan Provincial Engineering and Technological Research Center for Prevention and Treatment of Ophthalmology and Otolaryngology Diseases with Chinese Medicine and Protecting Visual Function,Changsha 410000,China;Xi’an Hospital of Traditional Chinese Medicine,Xi’an 710021,China;The First Affiliated Hospital of Hunan University of Chinese Medicine,Changsha 410007,China)

机构地区：[1]湖南中医药大学,长沙410208 [2]中医药防治眼病与视功能保护湖南省工程研究中心,长沙410208 [3]西安市中医医院,西安714000 [4]湖南中医药大学第一附属医院,长沙410007

出　　处：《中国中医基础医学杂志》2022年第3期370-374,共5页JOURNAL OF BASIC CHINESE MEDICINE

基　　金：国家自然科学基金资助项目(81774372)-破血消癥法对hRPE细胞增殖凋亡和MAPK、PI3K/AKT信号通路表达的影响;湖南省自然科学基金资助项目(2016JJ6111)-基于MAPK、PI3K/AKT信号途径研究散血明目片对PVR的作用机制;湖南省中医药管理局科研项目(201622)-散血明目片对hRPE细胞增殖及MAPK、PI3K/AKT信号通路的影响;湖南省科技创新平台与人才专项重点实验室和工程技术研发中心组项目(2017TP1018)-中医药防治五官科疾病湖南省重点实验室建设项目;湖南省中医药防治耳鼻喉疾病与视功能保护工程研究中心开放基金项目(2018YGCO4)-内服与外服相结合防治干眼的研究。

摘　　要：目的:观察活血散结方含药血浆对血小板源性生长因子(platelet derived growth factor,PDGF)干预下兔视网膜色素上皮(retinal pigment epithelium,RPE)细胞凋亡的影响以及对caspase-3、P53蛋白表达的干预作用。方法:采用高效液相色谱法检测活血散结方主要成分,选取第四代兔RPE细胞为实验细胞,分为空白对照组、正常血浆组、PDGF(10μg·L^(-1))组、PDGF(10μg·L^(-1))+AG1296(10μmol·L^(-1))组、PDGF(10μg·L^(-1))+10%含药血浆组、PDGF(10μg·L^(-1))+20%含药血浆组,分别加入相应处理因素干预48 h后,流式细胞仪检测兔RPE细胞凋亡情况,western blot检测兔RPE细胞caspase-3及P53蛋白表达情况。结果:活血散结方含药血浆中三七皂苷R1与丹参酮的含量均在线性范围之内,含药浓度较理想,活血散结含药血浆能促进PDGF干预下的兔RPE细胞凋亡(P<0.01),对caspase-3及P53蛋白表达亦有促进作用(P<0.01)。结论:活血散结中药复方含药血浆可能通过促进RPE细胞凋亡,达到抑制增殖性玻璃体视网膜病变发生发展的作用。Objective:To observe the effect of Huoxue Sanjie Recipe containing plasma on the apoptosis of rabbit retinal pigment epithelium(RPE)cells under PDGF intervention.Methods:The main components of Huoxue Sanjie Recipe containing plasma were detected by HPLC.The fourth generation of rabbit RPE cells were used as research subjects.According to respective processing factors,the subjects were divided into blank control group(DMEM),normal plasma group,PDGF(10μg·L^(-1))group,PDGF(10μg·L^(-1))+AG1296(10μmol·L^(-1))group,PDGF(10μg·L^(-1))+10%plasma containing drugs group,PDGF(10μg·L^(-1))+20%plasma containing drugs group.After 48 hours of intervention,the apoptosis of rabbit RPE cells was tested by flow cytometry,and the expression of caspase-3 and P53 protein in subjects were conducted by western blot.Results:The contents of notoginsenoside R1 and tanshinone in Huoxue Sanjie Recipe containing plasma were within the linear range,and both had ideal drug concentration.The medicated plasma can promote the apoptosis of rabbit RPE cells under PDGF intervention(P<0.01),and increase the expression of caspase-3 and P53 protein(P<0.01).Conclusions:Huoxue Sanjie Recipe containing plasma may inhibit the occurrence and development of PVR by promoting the apoptosis of RPE cells.

关 键 词：活血散结方 视网膜色素上皮细胞 凋亡 血小板源性生长因子 CASPASE-3 P53 

分 类 号：R285.5[医药卫生—中药学]