原花青素抑制同型半胱氨酸对人脐静脉内皮细胞的毒性作用

Grasp Seed Procyanidins Protecting Human Umbilical Vein Endothelial Cell From Homocysteine

目的探讨原花青素对同型半胱氨酸介导的人脐静脉内皮细胞毒性作用的影响及其作用的分子机制。方法采用MTT及乳酸脱氢酶检测细胞活性,流式细胞仪检测活性氧的变化,并用试剂盒检测原花青素对超氧化物歧化酶、谷胱甘肽过氧化物酶活性以及丙二醛浓度的影响。Western blot分析相关蛋白的表达与磷酸化水平。结果同型半胱氨酸处理细胞48 h显著降低细胞的存活率,升高细胞内活性氧水平与丙二醛含量,降低超氧化物歧化酶及谷胱甘肽过氧化物酶的活性;而预先给予不同浓度的原花青素处理细胞1 h,可提高细胞存活率,并可有效抑制细胞内活性氧水平与丙二醛含量的升高,维持超氧化物歧化酶、谷胱甘肽过氧化物酶的活性。原花青素还显著抑制同型半胱氨酸诱导的细胞外信号调节激酶、p38和c-Jun氨基末端激酶的磷酸化。用特异性抑制剂分析证实,同型半胱氨酸介导的细胞毒性是丝裂原活化蛋白激酶所依赖的。结论原花青素能抑制同型半胱氨酸介导的人脐静脉内皮细胞的毒性,其细胞保护作用可能是通过维持细胞内抗氧化酶活性,降低细胞内活性氧水平,最终阻断活性氧激活的丝裂原活化蛋白激酶信号通路。

Aim To investigate the protective effect of grasp seed procyanidins （CSP） on homocysteine （HCY）-induced cell viability in human umbilical vein endothelial cell （HUVEC） and related molecular mechanisms. Methods Cell viability was analyzed by using MTI＇ and lactate debydrogenase （LDH） assay. The levels of superoxide dismutase （SOD）, glutathione peroxidese （GSHPx） and malondialdebyde （MDA） were examined by kits and the level of intracellular reactive oxygen species （ROS） were determined by flow cytometric assay. Proteins and phosphorylation levels were examined by Western blot. Results Treatment of cultured HUVEC with HCY for 48 hours induced a significant decrease of cell viability. The levels of intracellular ROS and MDA were enhanced, and the activities of SOD and GSI-IPx were decreased. However, simultaneous treatment with GSP exhibited cytoprotective effects, reduced formation of the MDA and ROS, maintained the activity of SOD and GSHPx, prevented tho phoephorylation of exrtacellular sigeal regulated kinase （ ERK ）, p38 and c-Jun N-terminal kinase （ JNK）. Conclusion These results demonstrated that CSP had the protective capacity to antagonize HCY induced cell toxicity in HUVEC. The protective effect of C, SP may be partially dependent on an anti-oxidative stress effects and then inhibition of the activation of mitogen-activated protein kinase （MAPK）.