番茄红素对血管内皮细胞氧化应激损伤的作用及机制研究

The Protective Effects and Mechanism of Lycopene Against H_2O_2 induced Injury in Vascular Endothelial Cells

目的:观察番茄红素(lycopene,LYC)对于血管内皮细胞功能的作用,探讨其作用机制。方法:人脐静脉内皮细胞(HUVECs)处理实验分组:对照组,H2O2组,H2O2+LYC组(1、2、4、8μmolL-1)。MTT法检测HUVECs存活率;免疫印迹法(Western blot)检测p38MAPK蛋白磷酸化水平、抗凋亡蛋白B淋巴细胞/白血病-2(bcl-2)及线粒体凋亡通路相关蛋白bax的表达;细胞黏附能力测定和伤口愈合实验检测HUVECs粘附率和迁移率;TUNEL法检测HUVECs凋亡率;ELASA法测定HUVECs内活性氧(ROS),超氧化物歧化酶(SOD),乳酸盐脱氢酶(LDH)释放量和caspase-3的活性。结果:H2O2损伤后HUVECs存活率显著降低(P<0.01),凋亡率显著增加(P<0.01),黏附和迁移能力显著降低(P<0.01),bax和p-p38MAPK的表达上调,bcl-2的表达下调,并且ROS、LDH的释放和caspase-3的活性增加(P<0.01),SOD的释放减少。而LYC的预处理可以明显逆转H2O2以上作用。结论:H2O2氧化应激损伤中,LYC保护内皮细胞可能与其抗过氧化损伤细胞凋亡,抑制异常的p38MAPK信号通路有关。

Objective： To observe the effect oflycopene （LYC） in human umbilical endothelial cells （HUVECs） and investigate the mechanism of LYC. Methods： The groups of cells were divided as below： control group, H2O2 treatment group, H2O2 treatment ＋LYC （1、2、4、8 μmolL-1） treatment group. The survival rate of HUVECs was detected by MTT method; The indicator were detected by Western blot, which included the phosphorylation level of p38MAPK and the expression of mitochondrial apoptosis pathway proteins （bax）, B-lymphocyte / leukemia -2 （bcl-2） protein; The adhesive rate and migration of HUVECs was detected by the cellular adhesion ability assay and the wound healing assay; The apoptosis rate of HUVECs was detected by TUNEL; The indicator were detected by ELISA, which included the production of Reactive oxygen species （ROS）, superoxide dismutase （SOD）, lactate dehydrogenase （LDH） and caspase-3 activity. Results： After being treated with H2O2, the HUVECs survival rate was distinctly decreased （P〈0.01）, the HUVECs apoptosis rate was distinctly increased （P〈0.01）, the HUVECs adhesion and migration ability was distinctly reduced （P〈0.01）, The phosphorylation level of p38MAPK and the expression of bax protein was distinctly down-regulated （P〈0.01）, and the expression of bcl-2 protein was distinctly up-regulated （P〈0.01）, the production of ROS, LDH and caspase-3 activity was distinctly increased （P〈0.01）, the production of SOD was distinctly decreased （P〈0.01）. Moreover, after the treatment with LYC, these effects has been reversed. Conelusions： LYC can anti-oxidation stress injury induced by H202 of HUVECs via its capacity of anti-apoptosis and inhibition of abnormal p38MAPK signal pathway.