CAJ | 학술논문

目的探讨生存素(survivin,SVV)在白藜芦醇抗心肌微血管内皮细胞(cardiac microvascular endothelial cells,CMEC)缺血再灌注损伤中的作用.方法体外培养大鼠心肌微血管内皮细胞,建立缺血再灌注损伤模型,完全随机法分为正常对照组(对照组)、缺血再灌注组、缺血再灌注+白藜芦醇组(白藜芦醇组)、缺血再灌注+白藜芦醇+ LY294002组(LY294002组).采用MTT比色法检测CMEC的增殖数量,Transwell法检测CMEC的迁移数量,PI-AnnexinV检测CMEC的凋亡率.Western blot法检测细胞SVV的蛋白表达水平.结果缺血再灌注组CMEC增殖和迁移数量均显著低于对照组[0.19±0.03比0.42±0.07和(28±2)个/5个视野比(50±3)个/5个视野,P均＜0.01],凋亡率则显著高于对照组[(19.7±0.8)％比(5.4±0.3)％,P＜0.02].白藜芦醇组CMEC增殖和迁移数量均显著高于缺血再灌注组[0.36±0.07比0.19±0.03和(55±3)个/5个视野比(28±2)个/5个视野,P均＜0.05],凋亡率则显著低于缺血再灌注组[(9.6±0.7)％比(19.7±0.8)％,P＜0.05],SVV蛋白表达水平则高于缺血再灌注组(P＜0.05).而LY294002组CMEC增殖和迁移数量均显著低于白藜芦醇组[0.25±0.05比0.36±0.07和(34±3)个/5个视野比(55±3)个/5个视野,P均＜0.05],凋亡率则显著高于白藜芦醇组[(16.2±0.6)％比(9.6±0.7)％,P＜0.05],SVV蛋白表达水平低于白藜芦醇组(P＜0.05).结论白藜芦醇可抑制缺血再灌注诱导的心肌微血管内皮细胞凋亡,其机制可能与PI3K/Akt介导的SVV上调相关.
목적 탐토생존소(survivin,SVV)재백려호순항심기미혈관내피세포(cardiac microvascular endothelial cells,CMEC)결혈재관주손상중적작용.방법 체외배양대서심기미혈관내피세포,건립결혈재관주손상모형,완전수궤법분위정상대조조(대조조)、결혈재관주조、결혈재관주+백려호순조(백려호순조)、결혈재관주+백려호순+ LY294002조(LY294002조).채용MTT비색법검측CMEC적증식수량,Transwell법검측CMEC적천이수량,PI-AnnexinV검측CMEC적조망솔.Western blot법검측세포SVV적단백표체수평.결과 결혈재관주조CMEC증식화천이수량균현저저우대조조[0.19±0.03비0.42±0.07화(28±2)개/5개시야비(50±3)개/5개시야,P균＜0.01],조망솔칙현저고우대조조[(19.7±0.8)％비(5.4±0.3)％,P＜0.02].백려호순조CMEC증식화천이수량균현저고우결혈재관주조[0.36±0.07비0.19±0.03화(55±3)개/5개시야비(28±2)개/5개시야,P균＜0.05],조망솔칙현저저우결혈재관주조[(9.6±0.7)％비(19.7±0.8)％,P＜0.05],SVV단백표체수평칙고우결혈재관주조(P＜0.05).이LY294002조CMEC증식화천이수량균현저저우백려호순조[0.25±0.05비0.36±0.07화(34±3)개/5개시야비(55±3)개/5개시야,P균＜0.05],조망솔칙현저고우백려호순조[(16.2±0.6)％비(9.6±0.7)％,P＜0.05],SVV단백표체수평저우백려호순조(P＜0.05).결론 백려호순가억제결혈재관주유도적심기미혈관내피세포조망,기궤제가능여PI3K/Akt개도적SVV상조상관.
Objective To detect the role of surviving (SVV) in the protective effect of resveratrol against hypoxia/reperfusion injury (H/RI) of cardiac microvascular endothelial cells (CMECs).Methods CMECs isolated from the hearts of adult rats were exposed to hypoxia (94％ N2,5％ CO2,1％ O2) for 2 h followed by 4 h reoxygenation (95％ O2,5％ CO2).The cell proliferation of CMECs was measured by MTT assay and Transwell method was used to detect migration ability of CMEC,PI-AnnexinV double staining and flow cytometry technique were employed to observe the apoptotic rate of CMECs.The SVV protein expression was detected with Western blot method.Results Compared to control group,the proliferation (0.19 ± 0.03vs.0.42 ±0.07,P ＜0.01) and migration ((28 ± 2)/5HPF vs.(50 ±3)/5 HPF,P ＜0.01) abilities were impaired and the apoptosis index ((19.7 ± 0.8) ％ vs.(5.4 ± 0.3) ％,(P ＜ 0.05) of CMEC was increased after H/RI.The proliferation (0.36 ± 0.07 vs.0.19 ± 0.03,P ＜ 0.05) and migration ((55 ± 3)/5 HPF vs.(28 ± 2)/5 HPF,P ＜ 0.05) abilities of CMEC were significantly improved while the apoptosis index ((9.6 ±0.7) ％ vs.(19.7 ± 0.8) ％,P ＜ 0.05) was significantly decreased in H/RI + resveratrol group compared to H/RI group.SVV protein expression was also upregulated in H/RI + resveratrol group compared to H/RI group (P ＜0.05).To further ascertain the role of SVV in the protective effects of resveratrol,PI3K specific inhibitor LY294002 was added to H/RI + resveratrol group,the proliferation(0.25 ± 0.05 vs.0.36 ± 0.07,P ＜0.05) and migration ((34 ± 3)/5HPF vs.(55 ± 3)/5HPF,P ＜ 0.05) abilities were significantly decreased,the apoptosis index ((16.2 ± 0.6) ％ vs.(9.6 ± 0.7) ％,P ＜ 0.05) was increased and the protein expression of SVV was downregulated (P ＜ 0.05) in LY294002 + H/RI + resveratrol group compared to H/RI + resveratrol group.Conclusion Resveratrol could significantly reduce H/RI induced apoptosis and attenuate H/RI induced cardiac microvascular endothelial cells dysfunction through up-regulating PI3K/Akt/SVV pathways.