三羟异黄酮对胆固醇诱导内皮细胞损伤的保护作用

目的:探讨胆固醇(cholesterol)对人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVEC)凋亡、细胞增殖的影响,同时观察CuSO4溶液在其中的作用及三羟异黄酮(genistein,GST)干预HUVEC的损伤情况。方法:实验分为8组。对照组(等体积溶剂)、胆固醇低剂量组(25 mg/L)、中剂量组(50 mg/L)和高剂量组(100 mg/L);另外4组分别为以上各组加入终浓度为10μmol/L CuSO4溶液。应用流式细胞仪检测不同浓度胆固醇对细胞凋亡率的影响;应用四甲基偶氮唑蓝微量酶反应比色法(MTT试验)检测细胞的生存率。胆固醇损伤组(胆固醇50 mg/L+终浓度为10μmol/L的CuSO4溶液)分别加入GST 22.5μmol/L、45μmol/L和90μmol/L。结果:不同浓度的胆固醇可不同程度地诱导HUVEC细胞发生凋亡,抑制HUVEC细胞增殖,且随浓度增高,其作用均明显增强。各组加入CuSO₄溶液后可使其细胞凋亡率明显升高,增殖率下降。胆固醇损伤组的GSH-Px活力和NO生成量明显下降,丙二醛(MDA)含量升高,与正常对照组和不同剂量组比较差异均有统计学意义(P<0.05~P<0.01),而正常对照组和GST不同剂量组差异均无统计学意义(P>0.05)。结论:胆固醇可以诱导内皮细胞凋亡及抑制细胞增殖,CuSO₄溶液可促进胆固醇对HUVEC的损伤。GST对被胆固醇损伤的内皮细胞有一定的保护作用。

Role of genistein in protecting the damaged HUVEC induced by cholesterin

Objective: To discuss the influence of cholesterol on the apoptosis and proliferation of human umbilical vein endothelial cells,and to observe the role of CuSO₄ solution in the process and how the genistein affects the damage of HUVEC.Methods: The experiment was conducted in 8 groups:the comparison group(with equal volume of solvent),low dose of cholesterol group(25 mg/L),medium dose group(50 mg/L),high dose group(100 mg/L) and other four groups with the same content as the above groups,but CuSO₄ solvent was added to them with a concentration of 10 μmol/L,respectively.The influence of cholesterol at different consistency on the cell apoptosis was detected by flow cytometry,and the survival rate of the cells was detected by MTT test.When the cholesterol was in action(cholesterol 50 mg/L+final concentration 10 μmol/L CuSO₄ solution),genistein of 22.5 μmol/L,45 μmol/L and 90 μmol/L was added.Results: Different concentrations of cholesterol induced the apoptosis and suppressed the proliferation of HUVEC in different degrels.These effects were strengthened with the increasement of the dose.With the mixture of the CuSO₄ solution,the apoptosis rate had a remarkable increase and the survival rate decreased prominently.The group of cholesterol damage had a marked drop of GSH-Px vitality and NO generation,while the content of MDA increased with an obvious difference(P<0.05) compared with the group of normal and groups with different dosages.There was no big difference between the group of normal and those with different dossage of Genistein(P>0.05)Conclusions: The cholesterol can induce the apoptosis and inhibit the proliferation of HUVEC;CuSO₄ solution may promote the damage of cholesterol to HUVEC.Genistein can protect the damaged HUVEC induced by cholesterol to certain extent