Li Liu-xia, Li Xiu-fang, Zhang Lan-lan, Zhang Yi, Guo Rui-xia, Zhang Xiao-yan, Zhou Yan, Qiao Yu-huan, Su Ke
Department of Obstetrics and Gynecology, First Affiliated Hospital of Zhengzhou University, Zhengzhou, China.
Zhonghua Fu Chan Ke Za Zhi. 2013 Feb;48(2):102-6.
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.
Sixty patients with preeclampsia were recruited from October 2011 to June 2012 in the First Affiliated 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.
(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).
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.
探讨子痫前期患者血浆对人脐静脉内皮细胞（HUVEC）增殖和凋亡的影响，以及细胞损伤与溶血磷脂酸（LPA）受体之间的关系。
选取2011年10月至2012年6月在郑州大学第一附属医院就诊的60例子痫前期患者，其中30例为轻度子痫前期组，30例为重度子痫前期组。另选取30例健康孕妇作为健康孕妇组。检测三组血浆LPA水平。将HUVEC与三组血浆在体外共同培养，并设空白对照组。采用MTT法和流式细胞术检测HUVEC的增殖和凋亡情况。采用生物素链霉菌抗生物素蛋白过氧化物酶（SP）法免疫组化检测Edg 2、4、7蛋白表达水平。
（1）健康孕妇组、轻度子痫前期组和重度子痫前期组血浆LPA水平分别为（3.38±2.08）μmol/L、（6.12±0.22）μmol/L、（9.10±0.17）μmol/L。子痫前期患者血浆LPA水平显著高于健康孕妇（P<0.01）。（2）轻度和重度子痫前期组HUVEC增殖率分别为（65.2±2.7）%和（51.9±2.8）%，显著低于健康孕妇组和对照组[分别为（84.3±3.1）%和（100.0±0.0）%，P<0.01]。（3）轻度和重度子痫前期组HUVEC早期凋亡率、中晚期凋亡率及总凋亡率[总凋亡率分别为（30.4±2.0）%和（43.4±2.5）%]显著高于健康孕妇组和对照组[总凋亡率分别为（18.6±1.6）%和（8.0±1.5）%，P<0.01]。（4）四组Edg 2、4、7蛋白表达阳性率分别为：轻度子痫前期组83%、80%和73%；重度子痫前期组97%、93%和90%；健康孕妇组40%、40%和37%，对照组分别为10%、10%和7%。轻度和重度子痫前期组HUVEC阳性率显著高于健康孕妇组和对照组（P<0.01）。
子痫前期患者血浆可抑制HUVEC增殖并促进其凋亡，诱导Edg 2、4、7蛋白表达。提示血浆中溶血磷脂酸升高可能是子痫前期患者内皮细胞损伤的原因之一。
