摘要
目的观测子痫前期患者血清12-羟基二十碳四烯酸(12-HETE)浓度变化及对绒毛膜外滋养细胞诱导血管内皮细胞凋亡的影响,探讨12-HETE参与在子痫前期发生、发展的可能机制。方法收集2019年6月至2020年6月复旦大学附属中山医院吴淞医院子痫前期患者临床症状出现前26例和出现后23例,以及与之孕周、年龄及BMI相匹配的正常孕妇26例及正常未孕妇女14例血清样本,利用超高压液相色谱串联质谱仪(UPLC-MS)检测各组血清12-HETE浓度;建立人血管内皮细胞的单培养及其与胎盘滋养细胞HTR-8/Snveo的共培养模型,分为正常对照组、12-HETE处理组、共培养对照组、共培养12-HETE处理组。采用流式细胞术检测各组细胞血管内皮细胞的凋亡情况,计算凋亡率。结果12-HETE的保留时间为3.33 min,母离子碎片为319.33 m/z,离子碎片为179.22 m/z,在20-2000 pg/ml的线性范围内,相关系数r^(2)=0.99,能够满足样本检测需求。子痫前期患者临床症状出现前、后12-HETE浓度均高于正常孕妇组和正常未孕妇女组(均P<0.01)。血管内皮细胞单培养中,12-HETE处理组细胞凋亡率显著低于正常对照组(P<0.01);共培养对照组血管内皮细胞的凋亡率显著高于正常对照组(P<0.01);共培养12-HETE处理组血管内皮细胞的凋亡率显著低于共培养对照组(P<0.01)。结论子痫前期患者临床症状出现前后外周血中12-HETE浓度均显著升高,其可能通过抑制血管内皮细胞凋亡参与子痫前期的发生、发展。
Objective To investigate serum 12-hydroxyeicosatetraenoic acid(12-HETE)levels in preeclamptic patients and the effect of 12-HETE on apoptosis of endothelial cells co-cultured with trophoblasts.Methods Serum samples were collected from 26 pregnant women before(n=26)and after(n=23)onset of preeclampsia,26 age,gestational age and BMI matched normal pregnant women and 14 non-pregnant women in Wusong Hospital from June 2019 to June 2020.The serum 12-HETE levels were determined with ultra-high pressure liquid chromatography tandem mass spectrometer(UPLC-MS)in three groups.The human vascular endothelial EAHY926 cells were co-cultured with placental trophoblast HTR-8/Snveo cells,and treated or not treated(control group)with 12-HETE.Flow cytometry was used to detect the apoptosis of EAHY926 cells in the above groups.Results The retention time of 12-HETE in UPLC-MS was 3.33 min,the precursor ion fragment was 319.33 m/z,the ion fragment was 179.22 m/z,within the linear range of 20-2000pg/ml,the correlation coefficient(r^(2))was up to 0.99.The serum 12-HETE level in the preeclampsia group before and after preeclampsia onset was significantly higher than that of normal pregnant women and non-pregnant women(all P<0.01).In the single culture of vascular endothelial cells,the apoptosis rate of the 12-HETE group was significantly lower than that of the control group(P<0.01),in the cell co-culture,the apoptosis rate of vascular endothelial cells in the control group was significantly higher than that of the single culture control group(P<0.01).The apoptosis rate of vascular endothelial cells in the 12-HETE group was significantly lower than that in the control group(P<0.01).Conclusion The 12-HETE in the maternal peripheral blood before and after the clinical symptoms of preeclampsia patients are significantly increased,indicating that 12-HETE may be involved in the occurrence and development of preeclampsia by inhibiting the apoptosis of vascular endothelial cells.
作者
陈林坤
段玉萍
洪茂
袁想妹
厉倩
CHEN Linkun;DUAN Yuping;HONG Mao;YUAN Xiangmei;LI Qian(Department of Clinical Laboratory,Wusong Hospital,Zhongshan Hospital Affiliated Fudan University,Shanghai 200940,China)
出处
《浙江医学》
CAS
2021年第14期1513-1517,共5页
Zhejiang Medical Journal
基金
上海市宝山区科委科技创新专项(18-E-6)。
关键词
子痫前期
12-羟基二十碳四烯酸
凋亡
胎盘滋养细胞
血管内皮细胞
螺旋动脉重塑
Preeclampsia 12-hydroxyeicosatetraenoic acid
Apoptosis
Placental trophoblast cells
Vascular endothelial cells
Spiral artery remodeling
