紫花牡荆素抑制氧化应激诱导的HUVEC细胞凋亡

Protective Effect of Casticin on Apoptosis of HUVEC Cell Induced by Oxidative Stress

目的研究紫花牡荆素(CAS)对H2O2氧化应激诱导的HUVEC细胞凋亡的影响及其可能机制。方法体外培养HUVEC细胞,在添加H2O2氧化应激诱导之前先加入不同浓度的CAS(5、10和20μmoL/L)预孵30min,用MTT法观察各组细胞生长活性;用AO/EB染色和FCM法检测HUVEC细胞凋亡情况及其凋亡率;用Western blot检测细胞凋亡相关蛋白表达活性。结果 MTT提示与H2O2组相比较,CAS呈浓度和时间依赖性增加H2O2氧化应激诱导的HUVEC细胞的生长活性(P<0.05),降低HUVEC细胞的凋亡数量及凋亡率(P<0.05);同时下调Cytochrome C、Caspase-9和Caspase-3蛋白表达降低(P<0.05),激活Bcl-2蛋白表达(P<0.05),而Bax蛋白表达不变(P>0.05),Bcl-2/Bax上升(P<0.05)。结论 CAS拮抗H2O2氧化应激诱导的HUVEC细胞凋亡可能与其调节内源性线粒体凋亡途径有关。

Aim To investigate the protective effective of casticin（ CAS） against human umbilical vein endothelial cells（ HUVEC） apoptosis induced by H2O2oxidative stress and its possible molecular mechanism. Methods HUVEC cells were cultured in vitro. The cells which would be induced by H2O2were incubated in advance for 30 minutes with ONY and various concentration of CAS. Cells viability was measured by MTT assay,and its apoptosis-inducing effect were determined by AO / EB and FCM,meanwhile western blot assay was used to measure proteins related to apoptosis. Results CAS could increase the viability of HUVEC cells induced by H2O2in a dose and time-dependent manner compared with cells solo exposed to H2O2（ P〈0. 05）. The number of apoptotic cells and the apoptotic rate of HUVEC cells treated with various concentration CAS and H2O2significantly decreased in a dose and time-dependent manner compared with H2O2group（ P〈0. 05）,meanwhile the expression of Cytochrome-C,Caspase-9 and Caspase-3 protein were down-regulated（ P〈0. 05）,and protein level of Bcl-2 was activated（ P〈0. 05）,while the expression of Bax protein showed no change in the same treatment（ P〈0. 05）,and the ratio of Bcl-2 / Bax expression level increased（ P〈0. 05）. Conclusion CAS could prevent H2O2-induced HUVEC cells apoptosis which might be correlated with mitochondriacontrolled apoptotic pathway.