卡托普利与黄芪注射液对人脐静脉内皮细胞凋亡的影响

Effects of Captopril and Milkvetch Root Injection on Apoptosis of HUVEC Induced by H_2O_2

目的观察卡托普利、黄芪注射液及两者合用对过氧化氢(H2O2)诱导的人脐静脉内皮细胞(HUVEC)株凋亡的抑制作用,初步探讨其机制。方法将HUVEC分为11组:①正常对照组(Normal);②H2O2模型组(Model):细胞培养基中加入2.00mmol.L-1H2O2孵育4h;③卡托普利低、中、高浓度组:细胞培养基中加入0.5,5.0,50.0μg.mL-1卡托普利预处理24h,再加入2.00mmol.L-1H2O2孵育;④黄芪注射液低、中、高浓度组:细胞培养基中加入0.1,1.0,10.0mg.mL-1黄芪注射液预处理24h,再加入2.00mmol.L-1H2O2孵育;⑤两药合用低、中、高浓度组:细胞培养基同时加入具有上述3种终浓度卡托普利和黄芪注射液混合液预处理24h,再加入2.00mmol.L-1H2O2孵育。流式细胞术测定内皮细胞凋亡率,吖啶橙溴化乙啶荧光染色(AO-EB染色)观察细胞凋亡形态,Western blotting检测Bcl-2表达。结果与H2O2模型组比较,正常对照组、卡托普利各浓度组和黄芪注射液中、高浓度组早期凋亡率明显降低(P<0.05或P<0.01);正常对照组、卡托普利和黄芪注射液中、高浓度组总凋亡率均明显降低(P<0.05或P<0.01),Bcl-2含量明显升高(P<0.05或P<0.01)。各浓度联合用药组与单独用药组比较,早期凋亡率、总凋亡率明显降低,Bcl-2含量明显升高,均差异有显著性(P<0.05或P<0.01)。结论不同浓度卡托普利、黄芪注射液及两药合用可降低H2O2诱导的HUVEC凋亡率,增强Bcl-2表达,且呈一定浓度依赖性。相同浓度两药联用作用优于单药组。

Objective To investigate the protective effects and mechanisms of ACE1 （captopril） ,Milkvetch root and their combination on human umbilical vascular endothelial cells （HUVECs） damage induced by hydrogen peroxide（ H2O2 ）. Methods HUVEC were injuried for 4 hours by 2.00 mmol · L^-1 H2O2 after they were incubated with different dosages of eaptopril, Milkvetch root and their eombination for 24 hours. The morphology change of apoptosis was observed by AO/EB fluorescent staining. Apoptosis ratio of HUVEC was detected by FCM. Expression of Bel-2 protein was deteeted by Western blot technique. Results The early cell apoptosis ratio in the normal control and （captopril-L, M, H/Milkvetch root-M, H） were significantly lower than those in the H2O2 oxidative control （ P 〈 0. 05 or P 〈 0. 01 ）. The total cell apoptosis ratio in the normal control and （ captopril/Milkvetch root-M, H） ＋ H2O2 groups were significantly lower than those in the H202 oxidative control（ P 〈0.05 or P 〈 0.01 ） except captopril/Milkvetch root-L ＋ H2O2 group. Besides, the content of important inhibition apoptosis protein Bcl-2 of cell in the normal control and （captopril/Milkvetch root-M, H ） ＋ H2O2 groups were significantly higher than those in the H2O2 oxidative control （P 〈 0.05 or P 〈 0.01 ）. Significant differences were found between drug combination groups and single use groups in early apoptosis rate, total apoptosis rate and Bcl-2 content （ P 〈 0.05 or P 〈 0.01 ）. Conclusion Captopril, Milkvetch root and their combination can decrease the apoptosis ratio induced by H2O2, increase the level of Bcl-2 in a dose dependent way. The combined use of captopril and Milkvetch root injection is better than the single use of captopril and Milkvetch root.