Apelin对滋养细胞生物学功能的影响及对子痫前期样小鼠的治疗作用  

Effect of Apelin on the biological functions of trophoblast cells and its therapeutic effect on preeclampsia-like mice

作　　者：李梓汇 麻雨萌 王浩然 王宝茜 刘婕 毛海婷 Li Zihui;Ma Yumeng;Wang Haoran(Department of Clinical Laboratory,the Second Hospital of Shandong University,Jinan 250000)

机构地区：[1]山东大学第二医院检验医学中心,济南250000

出　　处：《现代妇产科进展》2024年第7期493-500,共8页Progress in Obstetrics and Gynecology

基　　金：济南市科技计划项目(No:202328046)。

摘　　要：目的:探讨Apelin对滋养细胞系生物学功能的影响及其对子痫前期(PE)样小鼠的治疗作用。方法:Western blot法检测正常妊娠者和PE患者胎盘组织中Apelin蛋白水平。RT-qPCR和Western blot法检测滋养细胞中Apelin mRNA和蛋白水平。细胞免疫荧光检测Apelin在滋养细胞中的定位情况。无标记细胞增殖实验、克隆形成实验以及EdU荧光染色检测Apelin对滋养细胞增殖能力的影响。细胞划痕实验和Transwell小室检测Apelin对滋养细胞迁移和侵袭能力的影响。血管形成实验检测Apelin对滋养细胞血管形成能力的影响。Western blot法检测对照组、不同浓度Apelin刺激组和Apelin抑制剂组基质金属蛋白酶2(MMP-2)、基质金属蛋白酶9(MMP-9)、N钙黏蛋白(N-cadherin)、E钙黏蛋白(E-cadherin)、Erk、P-Erk蛋白水平。构建PE样小鼠模型,并给予Apelin注射治疗,检测各组小鼠的收缩压和尿蛋白水平及小鼠的活胎比例、胎鼠重量、胎盘重量及二者比值。结果:PE患者胎盘组织中Apelin水平降低。3种滋养细胞中,Apelin均存在表达且定位于胞质。在细胞生物学功能方面,Apelin对滋养细胞的增殖能力无明显影响,但通过上调MMP-2和N-cadherin表达而促进滋养细胞的迁移和侵袭能力。血管形成实验证明,Apelin可促进滋养细胞的血管形成能力。Western blot法结果表明,Apelin对MAPK/ERK通路中Erk蛋白磷酸化水平有促进作用。PE小鼠模型的结果显示,Apelin可缓解其临床症状,并提高小鼠的胚胎存活率和发育水平。结论:Apelin在PE患者胎盘组织中表达降低,并可通过上调滋养细胞的迁移、侵袭和血管形成能力,对PE样小鼠具有治疗作用,可能成为PE的潜在治疗靶点。Objective:To explore the effect of Apelin on the biological functions of trophoblast cells and its therapeutic effect on preeclampsia-like mice.Methods:Western blot was used to detect the expression level of Apelin protein in placenta tissue of normal pregnant women and preeclampsia patients.RT-qPCR and Western blot were used to detect the expression levels of Apelin mRNA and protein in trophoblast cells.Cell immunofluorescence was used to detects the localization of Apelin in trophoblast cells.Label-free cell proliferation experiment,colony formation experiment and EdU staining were used to detect the effect of Apelin on the proliferation ability of trophoblast cells.Cell scratching and transwell chamber were used to detect the effect of Apelin on the migration and invasion ability of trophoblast cells.Blood vessel formation experiments were performed to detect the effect of Apelin on the blood vessel formation ability of trophoblast cells.Western blot was used to detect matrix metalloproteinase 2(MMP-2),matrix metalloproteinase 9(MMP-9),N-cadherin,E-cadherin,Erk,and P-Erk protein expression levels in the control group,Apelin stimulation group and Apelin inhibitor group.By constructing a preeclampsia-like mouse model and giving Apelin injection treatment,the systolic blood pressure and urine protein levels of mice in each group were detected,as well as the proportion of live fetuses,fetal weight,placental weight and the ratio between the two.Result:The expression of Apelin in the placental tissue of preeclampsia patients was reduced.Apelin was expressed and localized in the cytoplasm of trophoblasts.In terms of cell biological function,Apelin had no effect on the proliferation ability of trophoblasts,but promoted the migration and invasion ability of trophoblasts by up-regulating the expressions of MMP-2 and N-cadherin.Angiogenesis experiments proved that Apelin could promote the vascularization ability of trophoblasts.Western blot showed that Apelin promoted the phosphorylation level of Erk protein in the MAPK/

关 键 词：APELIN 子痫前期 滋养细胞 MAPK/ERK通路 迁移 血管形成 

分 类 号：R712.4[医药卫生—妇产科学]