探讨Notch1蛋白和过氧化物酶体增殖物激活受体(PPAR)-γ蛋白在早发型重度子痫前期(PE)孕妇胎盘组织中的表达及相关性。

选择2016年10月1日至2017年9月30日，在广东省妇幼保健院住院，行择期剖宫产术分娩的65例重度PE孕妇为研究对象。按照发病孕龄不同，将33例早发型重度PE孕妇(孕龄≤34孕周)纳入早发型重度PE组，将32例晚发型重度PE孕妇(孕龄>34孕周)纳入晚发型重度PE组。同时采用随机数字表法，随机选择同期在本院住院因胎位异常或者胎儿窘迫行择期剖宫产术分娩的30例孕妇，纳入对照组。采用免疫组织化学链霉菌抗生物素蛋白-过氧化物酶(SP)法检测Notch1和PPAR-γ蛋白在3组产妇胎盘组织中的分布及相对表达水平。采用单因素方差分析，对3组孕妇的年龄、孕龄和Notch1、PPAR-γ蛋白平均灰度值等进行整体比较，再采用最小显著差异(LSD)法进一步对3组孕妇Notch1和PPAR-γ蛋白平均灰度值进行两两比较。采用Pearson相关分析对孕妇胎盘组织中Notch1和PPAR-γ蛋白平均灰度值进行相关性分析。本研究遵循的程序符合广东省妇幼保健院人体试验委员会所制定的伦理学标准，并得到该伦理委员会审批(批准文号：201601027)，并且与所有研究对象均签署临床研究知情同意书。

①早发型重度PE组、晚发型重度PE组和对照组孕妇的年龄和孕龄等一般临床资料比较，差异均无统计学意义(P>0.05)。②Notch1与PPAR-γ蛋白在3组产妇的胎盘组织中均有表达，Notch1蛋白主要定位于胎盘滋养细胞的细胞质中，呈淡黄色-深棕色颗粒。PPAR-γ蛋白主要定位于绒毛细胞滋养层细胞核，少部分表达于细胞质内，呈黄褐色-深棕色颗粒。③3组孕妇胎盘组织中Notch1和PPAR-γ蛋白平均灰度值总体比较，差异均有统计学意义(F＝5.565，P＝0.012；F＝8.694，P＝0.006)。3组孕妇胎盘组织中Notch1蛋白平均灰度值进行两两比较，差异均有统计学意义(早发型重度PE组vs晚发型重度PE组：LSD-t＝3.218，P＝0.005；早发型重度PE组vs对照组：LSD-t＝4.832，P＝0.007；晚发型重度PE组vs对照组：LSD-t＝2.948，P＝0.008)。对3组孕妇胎盘组织中PPAR-γ蛋白平均灰度值分别进行两两比较，差异亦均有统计学意义(早发型重度PE组vs晚发型重度PE组：LSD-t＝3.412，P＝0.003；早发型重度PE组vs对照组：LSD-t＝4.927，P＝0.002；晚发型重度PE组vs对照组：LSD-t＝3.168，P＝0.001)。④在早发型重度PE组、晚发型重度PE组的胎盘组织中，Notch1和PPAR-γ蛋白的平均灰度值进行Pearson相关分析的结果显示，Notch1和PPAR-γ蛋白的平均灰度值呈负相关关系(r＝—0.461，P<0.05)。

早发型重度PE孕妇胎盘组织中Notch1相对表达水平下调，PPAR-γ蛋白相对表达水平上调，提示Notch1蛋白和PPAR-γ蛋白表达异常可能在滋养细胞的浸润过程中起着主导作用，并且影响早发型重度PE的发生、发展。

To investigate the expression and correlation of Notch1 protein and peroxisome proliferator-activated receptor (PPAR)-γ protein in placenta of early onset severe preeclampsia ( PE ) pregnant women.

From October 1, 2016 to September 30, 2017, 65 pregnant women with severe PE who were hospitalized in Guangdong Women and Children′s Hospital and underwent elective cesarean section were selected into this study. According to different gestational ages, 33 pregnant women with early onset severe PE (gestational age≤34 weeks) were included into early onset severe PE group, and 32 pregnant women with late onset severe PE (gestational age>34 weeks) were included in late onset severe PE group. At the same time, 30 pregnant women who were hospitalized in the same hospital due to abnormal fetal position or fetal distress and underwent elective cesarean section during the same period were randomly selected into control group. Immunohistochemically streptavidin peroxidase (SP) method was used to detect the distribution and expression levels of Notch1 protein and PPAR-γ protein in placental tissues. One-way ANOVA was used to compare the age, gestational age, average gray-value scale of Notch1 protein and PPAR-γ protein among three groups. The least significant difference (LSD) method was used to further compare the average gray-value scale of Notch1 protein and PPAR-γ protein among three groups. Pearson correlation analysis was used to analyze the correlation between Notch1 protein and PPAR-γ protein in placenta tissue of pregnant women. The procedures followed in this study were in accordance with the ethical standards established by the Human Subjects Trial Committee of Guangdong Women and Children′s Hospital, and this study was approved by this committee (Approval No. 201601027). Informed consent was obtained and signed with each subject.

①There were no significant differences in age and gestational age between early onset severe PE group, late onset severe PE group and control group (P>0.05). ② Notch1 protein was expressed in placental tissues of 3 groups, mainly localized in the cytoplasm of placental trophoblast cells, showing pale yellow-dark brown granules. PPAR-γ protein is mainly located in the nucleus of villous cytotrophoblast, and a small part of PPAR-γ protein is expressed in cytoplasm, presenting tan-dark brown granules. ③ The average grey-scale values of Notch1 protein and PPAR-γ protein in placental tissues among 3 groups were all statistically significant (F=5.565, P=0.012; F=8.694, P=0.006). Multiple comparison results showed the following results. The average gray-scale values of Notch1 protein in placenta tissues of 3 groups were respectively pairwise compared, and the differences were statistically significant (early onset severe PE group vs late onset severe PE group: LSD-t=3.218, P=0.005; early onset severe PE group vs control group: LSD-t=4.832, P=0.007; late onset severe PE group vs control group: LSD-t=2.948, P=0.008). The average grey-scale values of PPAR-γ protein in placental tissues were compared among 3 groups, and the differences were also statistically significant (early onset severe PE group vs late onset severe PE group: LSD-t=3.412, P=0.003; early onset severe PE group vs control group: LSD-t=4.927, P=0.002; late onset severe PE group vs control group: LSD-t=3.168, P=0.001). ④Pearson correlation analysis of Notch1 protein and PPAR-γ protein in placenta tissues of early severe PE group and late severe PE group showed that the average gray-scale values of Notch1 protein and PPAR-γ protein were negatively correlated (r=-0.461, P<0.05).

The relative expression level of Notch1 and PPAR-γ protein in placental tissue of pregnant women with early onset severe PE were down-regulated and up-regulated, suggesting that abnormal expressions of Notch1 protein and PPAR-γ protein may play a leading role in the infiltration of trophoblasts and affect the occurrence and development of early onset severe PE.