复方银杏叶颗粒改善人脐静脉内皮细胞氧化应激损伤作用及其机制研究.docVIP

复方银杏叶颗粒改善人脐静脉内皮细胞氧化应激损伤作用及其机制研究 　　[摘要]该实验于体外使用人脐静脉内皮细胞（HUVEC）与复方银杏叶颗粒（80，160，320，640 mg?L-1）预孵育，合并使用H2O2（1 200 μmol?L-1）建立氧化应激损伤模型。采用MTT法研究药物对HUVEC细胞增殖情况的影响，检测细胞培养上清中的LDH，MDA，NO含量及SOD活性，判断药物对内皮细胞的保护作用。通过Casepase-3活性检测以及AnnexinV-FITC/PI流式细胞术，检测药物对细胞凋亡的保护作用。使用 Western blot法测定凋亡相关蛋白Bcl-2和Bax的表达。研究发现1 200 μmol?L-1H2O2能够诱导内皮细胞产生氧化应激损伤，使细胞存活率降低，且细胞增殖抑制程度与H2O2的作用时间呈正相关。而80，160，320，640 mg?L-1的复方银杏叶颗粒能够明显减轻该模型下内皮细胞的氧化应激损伤，恢复细胞正常增殖水平，改善细胞状态，抑制细胞凋亡，同时能够上调Bcl-2的表达以及下调Bax的蛋白表达。该实验证明了复方银杏叶颗粒对H2O2诱导的内皮细胞氧化应激损伤及细胞凋亡具有一定的保护作用，其机制可能与复方银杏叶颗粒抑制了内皮细胞凋亡的线粒体途径有关。 　　[关键词]复方银杏叶颗粒；H2O2；HUVEC；内皮细胞损伤；细胞凋亡 　　[Abstract]To reveal the protective and anti-apoptosis effect of compound Ginkgo biloba granules on oxidative stress injury of human umbilical vein endothelial cells （HUVEC）. Negative control group， H2O2 model group and 4 drug pretreatment groups （80， 160， 320， 640 mg? L-1） were established. The cell proliferation， morphological changes in each group after oxidative stress injury was detected by MTT assay and through microscope observation respectively. The content of LDH， MDA， SOD and NO and SOD activity in supernatant were detected to judge the protection effect of the drugs on endothelial cells. The protective effect on HUVEC apoptosis was analyzed by Caspase-3 activity test and Annexin V-FITC/PI staining. Western blot was used to observe the expression of apoptosis-related proteins Bcl-2 and Bax. Results showed that 1 200 μmol? L-1H2O2 can induce oxidative stress injury in endothelial cells and reduce the cell survival rate； cell proliferation inhibition degree is positively correlated with the effect time of H2O2. Besides， 80， 160， 320 640 mg?L-1compound Ginkgo biloba granules can protect HUVEC from oxidative stress injury， recover the normal proliferation level of cells， improve their state， prohibit cell apoptosis， and can up-regulate and down-regulate the expression level of Bcl-2 and Bax respectively. In conclusion， compound G. biloba granules can protect HUVEC from the oxidativ