Objectives To investigate the effects of plasma from the patients with preeclampsia on proliferation and apoptosis of human umbilical vein endothelial cells(HUVEC),and to explore the relationship between cell damage and lysophosphatidic acid(LPA)receptors.Methods Sixty patients with preeclampsia were recruited from October 2011 to June 2012 in the First Affilated Hospital of Zhengzhou University.Among them,thirty cases were defined as the mild preeclampsia group and thirty cases were defined as the severe preeclampsia group.The other thirty healthy pregnant women were recruited in the healthy pregnant women group.The levels of plasma LPA in the three groups were measured.The HUVEC were cultured in vitro with plasma from the three groups,and a blank control group was set up as well.Proliferation and apoptosis of HUVEC were measured by MTT assay and flow cytometry.Immunohistochemistry of biotin streptomyces protein peroxidase(SP)method was used to measure the protein expression level of Edg 2,4,7.Results(1)The plasma LPA levels in the healthy pregnant woman group,mild preeclampsia group and severe preeclampsia group were(3.38 ± 2.08)μmol/L,(6.12 ± 0.22)μmol/L,(9.10 ± 0.17)μmol/L,respectively.The plasma levels of LPA in patients with preeclampsia were significantly higher than that in the healthy pregnant women(P ＜ 0.01).(2)The proliferation rate of HUVEC in the mild and severe preeclampsia groups [(65.2 ± 2.7)％ and(51.9 ± 2.8)％] were significantly lower than that in the healthy pregnant women group and the control group [(84.3 ± 3.1)％ and(100.0 ± 0.0)％,P ＜ 0.01].(3)The early apoptosis rate,middle-late apoptosis rate and total apoptosis rate of HUVEC in the mild and severe preeclampsia groups [total apoptosis rate were(30.4 ±2.0)％ and(43.4 ±2.5)％] were significantly higher than those in the healthy pregnant women group and the control group [total apoptosis rate were(18.6 ± 1.6)％ and(8.0 ± 1.5)％,P ＜ 0.01].(4)The expression positive rates of Edg 2,4,7 proteins in the four groups were as following:mild preeclampsia group 83％,80％ and 73％;severe preeclampsia group 97％,93％ and 90％;healthy pregnant women group 40％,40％ and 37％,and the control group 10％,10％ and 7％ respectively.The positive rates of HUVEC in the mild and severe preeclampsia groups were significantly higher than those in the healthy pregnant women group and the control group(P ＜ 0.01).Conclusions The plasma of patients with preeclampsia could inhibit proliferation and promote apoptosis of HUVEC,and induce the expression of Edg 2,4,7 proteins.It suggested that the increase of lysophosphatidic acid in plasma could be one of the reasons of endothelial cell damage in patients with preeclampsia.%目的 通过观察子痫前期孕妇血浆对人脐静脉内皮细胞(HUVEC)增殖和凋亡的作用,探讨子痫前期孕妇血浆诱导HUVEC损伤与溶血磷脂酸(LPA)及其受体的关系.方法 选择2011年10月-2012年6月郑州大学第一附属医院子痫前期孕妇60例,分为轻度子痫前期组30例,重度子痫前期组30例.另选取同期入院分娩的健康孕妇30例为健康孕妇组.采用生化法测定各组孕妇血浆LPA水平；对体外培养的HUVEC中加入各组孕妇血浆进行培养,另设一个空白对照组.应用四甲基偶氮唑蓝比色法和流式细胞仪检测各组细胞的增殖及凋亡情况；免疫组化SP法检测各组细胞中内皮分化基因(Edg)2、4、7蛋白的表达.结果(1)健康孕妇组血浆LPA水平为(3.38±2.08)μmol/L,轻度子痫前期组为(6.12±0.22)μmol/L,重度子痫前期组为(9.10±0.17)μmol/L.轻、重度子痫前期组孕妇血浆LPA水平明显高于健康孕妇组,分别比较,差异均有统计学意义(P＜0.01).(2)轻、重度子痫前期组孕妇HUVEC增殖率[分别为(65.2±2.7)、(51.9±2.8)％]明显低于健康孕妇组和对照组[分别为(84.3±3.1)、(100.0±0.0)％],分别比较,差异均有统计学意义(P＜0.01).(3)轻、重度子痫前期组HUVEC早期凋亡率、中晚期凋亡率以及总凋亡率[总凋亡率分别为(30.4±2.0)％及(43.4±2.5)％]均明显高于健康孕妇及空白对照组[总凋亡率分别为(18.6±1.6)％及(8.0±1.5)％],分别比较,差异均有统计学意义(P＜0.01).(4)轻度子痫前期组HUVEC中Edg2、4、7蛋白表达阳性率分别为83％、80％及73％,重度子痫前期组分别为97％、93％及90％,健康孕妇组分别为40％、40％及37％,空白对照组分别为10％、10％及7％,轻、重度子痫前期组Edg2、4、7蛋白表达阳性率均明显高于健康孕妇组和空白对照组,分别比较,差异均有统计学意义(P＜0.01).结论 子痫前期孕妇血浆有抑制HUVEC增殖、促进其凋亡的作用,并诱导HUVEC中Edg 2、4、7蛋白表达.提示子痫前期孕妇血浆中LPA水平升高可能是血管内皮细胞损伤的原因之一.
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