miR-150在胎盘表达与复发性流产的关系研究

目的探讨miR-150通过靶向血管内皮生长因子(VEGF)表达调控胎盘血管生成参与复发性流产的关系。方法选择华中科技大学同济医学院附属梨园医院妇产科单胎晚期妊娠分娩的产妇40例为对照组(年龄23~29岁,平均年龄26.72岁),同期选择于该院妇产科3次或更多次自然流产的妇女40例为病例组(年龄23~29岁,平均年龄26.18岁)。测定两组胎盘组织miR-150 mRNA表达水平及VEGF表达水平。设立HTR-8/SVNEO细胞组、miR-150mimics组、miR-150 inhibitor组,测定3组细胞活力、凋亡水平、血管形成水平、miR-150 mRNA表达水平及VEGF表达水平。建立正常小鼠组、miR-150 mimics小鼠组、miR-150 inhibitor小鼠组模型,测定3组小鼠平均流产数、胎鼠质量、胎盘质量、胎盘毛细血管数目、miR-150 mRNA及VEGF表达水平。结果病例组胎盘组织miR-150 mRNA表达水平高于对照组胎盘组织,VEGF表达水平低于对照组胎盘组织(P <0.05)。miR-150 mimics组光密度(OD)值、细胞存活率、相对总血管长度、相对总血管数目、VEGF表达水平低于HTR-8/SVNEO细胞组(P <0.05);miR-150 inhibitor组OD值、细胞存活率、相对总血管长度、相对总血管数目、VEGF表达水平高于HTR-8/SVNEO细胞组和miR-150 mimics组(P <0.05)。miR-150 mimics组细胞凋亡率、miR-150 mRNA表达水平高于HTR-8/SVNEO细胞组(P <0.05),miR-150 inhibitor组细胞凋亡率、miR-150 mRNA表达水平低于HTR-8/SVNEO细胞组和miR-150 mimics组(P <0.05)。miR-150 mimics小鼠组平均流产数、miR-150 mRNA水平高于正常小鼠组(P <0.05),胎鼠质量、胎盘质量低于正常小鼠组(P <0.05)。miR-150inhibitor小鼠组平均流产数、miR-150 mRNA水平低于正常小鼠组和miR-150 mimics小鼠组(P <0.05),胎鼠质量、胎盘质量高于正常小鼠组和miR-150 mimics小鼠组(P <0.05)。miR-150 mimics小鼠组平均胎盘毛细血管数目水平、VEGF表达水平低于正常小鼠组(P <0.05),miR-150 inhibitor小鼠组平均胎盘毛细血管数目、VEGF表达水平高于正常小鼠组和miR-150 mimics小鼠组(P <0.05)。结论复发性流产患者胎盘组织中miR-150表达水平升高,VEGF表达水平降低;miR-150靶向抑制胎盘中VEGF表达水平进而抑制胎盘中血管的生成可能是复发性流产的发病机制之一。

Relationship of mi R-150 expression in placenta and recurrent spontaneous abortion

Objective To investigate the effect of miR-150 in regulation of placental angiogenesis by targeting the expression of vascular endothelial growth factor(VEGF) in recurrent spontaneous abortion. Methods A total of 40 singleton parturient women of late pregnancy were selected as control group(aged 23-29 years old with mean age of 26.72 years old), at the same time, 3 or more spontaneous abortion 40 females were set as case group(aged 23-29 years old with mean age of 26.18 years old).The expression level of miR-150 and VEGF in placenta tissue of 2 groups were measured. The HTR-8/SVNEO cell group,miR-150 mimics group and miR-150 inhibitor group were established, and cell viability, apoptotic level, angiogenesis level,expression of miR-150 mRNA and VEGF in 3 groups was measured. Normal mice group, miR-150 mimics mice group and miR-150 inhibitor mice group were established. The mean abortion number, fetal body weight, placenta weight, placental capillaries number, expression of miR-150 mRNA and VEGF in 3 groups were determined and compared. Results The expression level of miR-150 mRNA in placental tissue of case group was higher than that of control group, and the VEGF expression level was lower than that of control group(P < 0.05). The optical density(OD) value, survival rate, relative total blood vessel length,relative total blood vessel number and VEGF expression level of miR-150 mimics group were lower than those of HTR-8/SVNEO cell group(P < 0.05). The apoptosis rate and miR-150 mRNA expression of miR-150 mimics group were higher than those of HTR-8/SVNEO cells group(P < 0.05). The apoptosis rate and miR-150 mRNA expression level in miR-150 inhibitor group were lower than those in HTR-8/SVNEO cell group and miR-150 mimics group(P < 0.05). The mean abortion number and miR-150 mRNA level in miR-150 mimics group were higher than those in normal mice group(P < 0.05), while the fetal body weight and placental weight were lighter than those in normal mice group(P < 0.05). The mean abortion number and miR-150 mRNA level in miR-150 inhibitor mice group were lower than those in normal mice group and miR-150 mimics mice group(P < 0.05), while fetal body weight and placental weight were higher than those in normal mice group and miR-150 mimics mice group(P < 0.05). The mean placental capillary count and VEGF expression in miR-150 mimics group were lower than those in normal mice group(P < 0.05). The mean number of placental capillaries and VEGF expression in miR-150 inhibitor group were higher than those in normal mice group and miR-150 mimics group(P < 0.05). Conclusion It is demonstrated that the expression of microRNA-150 increases and VEGF expression decreases in placenta recurrent abortion patients. It may be one of pathogenesis in recurrent abortion that miR-150 targeting inhibits VEGF expression in placenta and then inhibits the angiogenesis in placenta.