摘要
目的探讨白藜芦醇在抑制缺氧/复氧诱导心肌微血管内皮细胞凋亡中的作用机制。方法体外培养大鼠心肌微血管内皮细胞,建立细胞缺氧/复氧损伤模型,随机分为正常对照组、缺氧/复氧组(H/R,2h/4h)、缺氧/复氧+白藜芦醇组(H/R+Res)。白藜芦醇分别选择5、10、20、30、40μmol/L浓度,通过TUNEL法检测心肌微血管内皮细胞的凋亡率以确定药物的最适浓度,确定最适浓度后进行后续实验,分组:正常对照组、缺氧/复氧组(H/R,2h/4h)、H/R+白藜芦醇组(最适浓度20μmol/L)、H/R+白藜芦醇(最适浓度20μmol/L)+LY294002(20μmol/L)组。MTT比色法检测细胞的增殖能力,Transwell法检测细胞迁移能力,流式细胞仪检测细胞的凋亡率。结果与缺氧/复氧组(H/R,2h/4h)相比,不同浓度的白藜芦醇可抑制细胞凋亡,并呈剂量依赖性,20μmol/L白藜芦醇组出现明显保护作用(P<0.01);LY294002处理后,细胞的增殖及迁移能力明显减弱(P<0.05,P<0.01),细胞凋亡率明显升高(P<0.01)。结论白藜芦醇可减少缺氧/复氧诱导的心肌微血管内皮细胞凋亡,其机制可能与通过PI3K/Akt信号通路表达有关。
Objective To investigate the effects of resveratrol on hypoxia/reoxygenation-induced apoptosis of cardiac microvascular endothelial cells(CMECs)and possible mechanisms.Methods CMECs were isolated from rat left ventricular by enzyme digestion and cultured in vitro.H/R injury model of CMECs in vitro was established.Collected CMECs were randomized into three groups:normal control group,hypoxia/reoxygenation Group(H/R,2 h/4 h),H/R +Resveratrol with different concentrations(5,10,20,30,40 μmol/L).The apoptotic rate of CMECs was detected by TUNEL colorimetric detection to determine the optimal dose of resveratrol(20 μmol/L)group,H/R group(2 h/4 h),H/R+resveratrol(20 μmol/L)group and H/R+resveratrol(20 μmol/L)+antagonist LY294002 group.MTT colorimetric assay was performed to detect CMECs proliferation,Transwell method was used to detect CMECs migration,AnnexinV/PI double staining and flow cytometry technique were employed to observe CMECs apoptotic rate.Results The resveratrol could significantly ameliorate CMECs apoptosis induced by H/R(2 h/4 h)pretreatment,and the optimal dose was 20 μmol/L(P<0.01);while treated with LY294002,the protective role of resveratrol on CMECs apoptosis was decreased significantly,apoptosis rate increased significantly(P<0.01),resveratrol on cell proliferation and migration of decreased significantly(P<0.05,P<0.01),significantly elevated apoptosis index(P<0.01).Conclusions Resveratrol can decrease hypoxia/reoxygenation-induced cardiac microvascular endothelial cells of apoptosis,and this may be related with activation of PI3K/Akt signaling pathway.
出处
《中华临床医师杂志（电子版）》
CAS
2012年第20期83-85,共3页
Chinese Journal of Clinicians(Electronic Edition)
