下调lncRNA-H19表达对H_(2)O_(2)诱导的HUVEC炎性损伤的影响  

Effects of lncRNA-H19 down-regulation on H_(2)O_(2)-induced inflammatory injury in HUVEC

作　　者：宋姗姗[1] 杨洋 陈阳[1] 何静[1] SONG Shanshan;YANG Yang;CHEN Yang;HE Jing(Department of Obstetrics and Gynecology,First Affiliated Hospital of Xi’an Medical College,Xi’an 710077,China;Department of Reproductive Medicine Center,Xi’an Peoples Hospital,Xi’an Fourth Hospital)

机构地区：[1]西安医学院第一附属医院产科,西安710077 [2]西安市人民医院,西安市第四医院生殖医学中心

出　　处：《山西医科大学学报》2022年第9期1150-1156,共7页Journal of Shanxi Medical University

基　　金：陕西省自然科学基础研究计划项目(2020JM-615)。

摘　　要：目的分析子痫前期(preeclampsia,PE)患者胎盘及血清中长链非编码RNA(lncRNA)H19表达情况,及抑制其表达对过氧化氢(H_(2)O_(2))诱导的人脐静脉内皮细胞(human umbilical vascular endothelial cell,HUVEC)炎性损伤的影响。方法收集15例PE晚孕患者(PE组)与15例正常晚孕患者(正常组)血清及胎盘组织标本,荧光实时定量聚合酶链式反应(quantitative real time polymerase chain reaction,qRT-PCR)检测两组血清及胎盘组织中lncRNA-H19的表达。以转染了siRNA-H19的人脐静脉内皮细胞(human umbilical vascular endothelial cell,HUVEC)及未转染的HUVEC为研究对象,根据是否进行H_(2)O_(2)干预培养分为4组:NC组(阴性对照)、H_(2)O_(2)组(H_(2)O_(2)干预培养)、H_(2)O_(2)+H19抑制对照组(转染siRNA-H19阴性对照物+H_(2)O_(2))、H_(2)O_(2)+H19抑制组(转染siRNA-H19+H_(2)O_(2))。qRT-PCR检测各组中lncRNA-H19表达。酶联免疫吸附法(ELISA)检测各组HUVEC培养上清中白细胞介素18(IL-18)、白细胞介素1β(IL-1β)表达。蛋白免疫印迹(Western blot)检测4组HUVEC中磷酸化细胞外调节蛋白激酶(phosphorylated extracellular regulated protein kinases,p-ERK)、磷酸化c-Jun氨基末端激酶(phosphorylated c-Jun N-terminal kinase,p-JNK)和磷酸化的p38丝裂原活化蛋白激酶(phosphorylated p38 mitogen activated protein kinase,p-p38)表达。管样形成实验检测4组HUVEC成管能力(管腔数及总分支长度);流式细胞术检测4组HUVEC凋亡能力。结果与正常组相比,PE组患者血清及胎盘组织中lncRNA-H19的表达降低(均P<0.05)。与其他3组相比,H_(2)O_(2)+H19抑制组HUVEC中lncRNA-H19表达降低(P<0.05)。4组中H_(2)O_(2)+H19抑制组上清液中IL-1β、IL-18、p-ERK、p-JNK、p-p38表达水平最高,HUVEC管腔数及总分支长度最低,HUVEC凋亡率最高(均P<0.05)。结论lncRNA-H19在PE患者血清及胎盘组织中低表达,下调lncRNA-H19能够进一步加重H_(2)O_(2)诱导的HUVEC细胞炎性损伤。Objective To investigate the expression of long-chain noncoding RNA(lncRNA)H19 in placenta and serum of patients with preeclampsia(PE),and its effect on the inflammatory injury of human umbilical vein endothelial cell(HUVEC)induced by hydrogen peroxide(H_(2)O_(2)).Methods Serum and placental tissues were collected from 15 late-pregnant PE patients(PE group)and 15 normal late-pregnant controls(normal group),and the expression of lncRNA-H19 in serum and placental tissue in the two groups was detected by fluorescence real-time quantitative polymerase chain reaction(qRT-PCR).Human umbilical vascular endothelial cells(HUVECs)were divided into four groups:NC group(negative control),H_(2)O_(2)group,H_(2)O_(2)+H19 inhibition control group(siRNA-H19 negative control transfection and H_(2)O_(2)intervention)and H_(2)O_(2)+H19 inhibition group(siRNA-H19 transfection and H_(2)O_(2)intervention).RT-qPCR was used to detect the expression of lncRNA-H19 in each group.The expression levels of interleukin-18(IL-18)and interleukin-1β(IL-1β)in HUVEC culture supernatant were detected by enzyme-linked immunosorbent assay(ELISA).The expression levels of phosphorylated extracellular regulated protein kinases(p-ERK),phosphorylated c-Jun N-terminal kinase(p-JNK)and phosphorylated p38 mitogen activated protein kinase(p-p38)were detected by Western blot.Tube-like formation assay was used to detect the tube-forming ability(number of lumens and total branch length)of HUVEC in four groups,and flow cytometry was used to detect the apoptosis ability of HUVEC in four groups.Results Compared with normal group,the expression of lncRNA-H19 in serum and placenta was decreased in PE group(all P<0.05).Compared with the other three groups,the expression of lncRNA-H19 in HUVEC was decreased in H_(2)O_(2)+H19 inhibition group(P<0.05).Among the four groups,the expression levels of IL-1β,IL-18,p-ERK,p-JNK and p-p38 in the supernatant were the highest in H_(2)O_(2)+H19 inhibition group,the number of HUVEC lumen and total branch length were the lowest,and t

关 键 词：长链非编码RNA 炎性损伤 子痫前期 人脐静脉内皮细胞 凋亡率 

分 类 号：R714.24[医药卫生—妇产科学]