Genistein对子痫前期患者离体脐静脉内皮细胞的保护作用及其机制研究

Effect of genistein on the number and function of HUVECs in preeclampsia patients and its mechanisms

目的分析染料木素(Genistein)对子痫前期患者脐静脉内皮细胞(HUVECs)数量及功能变化的影响及其机制。方法选取同期住院、年龄孕周及病变程度无显著差异的子痫前期患者40例,随机分成Genistein干预组及患者组(每组20例),20例正常妊娠孕妇为对照组,分离并培养上述研究对象的HUVECs,采用MTT等方法检测3组内皮细胞的增殖、黏附和迁移能力,采用荧光标记的牛血清白蛋白(BSA)检测3组内皮细胞的单层屏障功能;同时采用ELISA法检测内皮细胞8-羟基脱氧鸟苷(8-OHdG)水平,比色法检测谷胱甘肽过氧化物酶(GSH-Px)水平,Real-time PCR和Western blot法检测凋亡相关的Bcl-2、Bax基因和蛋白表达的水平。结果与子痫前期患者组比较,Genistein干预组内皮细胞增殖、黏附、迁移能力及其屏障功能明显升高,内皮细胞GSH-Px活性、抗凋亡的Bcl-2基因和蛋白表达水平明显升高,同时,8-OHdG水平及促凋亡的Bax基因和蛋白表达水平明显下降,且差异均有显著性(P<0.01)。而Genistein干预组与正常妊娠组比较,上述差异无显著性。结论 Genistein可能通过抗氧化、抗凋亡,减少子痫前期患者内皮细胞的损伤,提高内皮细胞的增殖能力及屏障功能,从而对子痫前期的发生、发展起预防和治疗的作用。

Objective To study the effect of genistein on endothelial cell damage in preeclampsia.Methods Human umbilical vein cells(HUVECs)were isolated from 20 normal pregnant women(control group)and 40 preeclampsia patients.Half of cells from the preeclampsia patients were treated with genistein(PE+Gen group)and the other half were untreated(PE group).HUVEC barrier function and proliferation were investigated using bovine serum albumin and MTT assays,respectively.Markers for oxidative stress and apoptosis were also assessed.Results Compared to the PE group,HUVEC barrier function,proliferation,glutathione peroxidase activity,and Bcl-2 gene and protein expression were significantly increased in the genistein treatment group(PE+Gen),whereas 8-hydroxy-2’-deoxyguanosine levels and Bax gene and protein expression were significantly decreased(P<0.01).There was no significant difference between PE+Gen and control groups.Conclusion Genistein reduces endothelial cell damage possibly through antioxidant and anti-apoptotic mechanisms,and it may be useful in preeclampsia prevention and treatment.