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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 200μmol·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Annexin V-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 200μmol·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。
The experiment in vitro using human umbilical vein endothelial cells (HUVEC) and Compound Ginkgo biloba particles (80,160,320,640 mg · L-1) pre-incubated with H2O2 (1,200 μmol·L-1) to establish a model of oxidative stress injury. The effects of drugs on the proliferation of HUVECs were studied by MTT assay. The contents of LDH, MDA, NO and SOD in the cell culture supernatants were detected to determine the protective effects of the drugs on endothelial cells. CasePase-3 activity assay and Annexin V-FITC / PI flow cytometry to detect the protective effect of the drug on apoptosis. Western blot was used to detect the expression of Bcl-2 and Bax. The study found that 1 200μmol·L-1H2O2 can induce endothelial cell oxidative stress injury, so that cell survival rate decreased, and the degree of inhibition of cell proliferation and duration of H2O2 was positively correlated. The compound Ginkgo biloba leaves of 80, 160, 320 and 640 mg · L-1 could significantly reduce the oxidative stress injury of endothelial cells in this model, restore the normal proliferation of cells, improve the cell status, inhibit the apoptosis of cells, and up-regulate the expression of Bcl-2 Down-regulate Bax protein expression. This experiment proves that compound Ginkgo biloba leaves have a protective effect on H2O2-induced endothelial cell oxidative stress injury and apoptosis, and its mechanism may be related to mitochondrial pathway that Compound Ginkgo biloba leaves inhibit endothelial cell apoptosis.