Xu Tingting, Chen Daijuan, Deng Xixi, Zhan Yongchi, Zhou Fan, Wang Xiaodong
Department of Obstetrics and Gynecology, West China Second University Hospital, Sichuan University, Sichuan 610041, China.
Key Laboratory of Birth Defects and Related Diseases of Women and Children (Sichuan University), Ministry of Education, Sichuan 610041, China.
Matern Fetal Med. 2021 Dec 15;4(1):7-16. doi: 10.1097/FM9.0000000000000137. eCollection 2022 Jan.
To investigate the possible regulatory mechanism of corticotropin-releasing hormone (CRH), urocortin (UCN), and Wolfram syndrome 1 (WFS1) in 17α-ethynylestradiol (EE)-induced intrahepatic cholestasis pregnant rats and its ischemia reperfusion (IR) model.
Pregnant rats ( = 60) were randomly divided into four experimental groups by random number table (Control, EE, IR, and EE-IR groups), and were studied on the 17, 19, and 21 gestational days (GD) ( = 5 in each group at the indicated time). Growth and development indicators of fetal rats among these four groups were recorded. Enzyme-linked immunosorbent assay was employed to detect CRH, UCN, and WFS1 levels in maternal sera. Western blotting and real-time polymerase chain reaction were used to quantify placental protein and placental mRNA levels of CRH, UCN, and WFS1. Multivariate analysis of variance and least significant difference test were used to establish the group and individual comparisons.
A significant difference was found in placenta weight ( = 8.10, < 0.05), fetal rat weight ( = 40.86, < 0.05), fetal rat length ( = 61.61, < 0.05), and fetal rat tail length ( = 55.63, < 0.05) among four groups on the 17 ,19 , and 21 GD.What's more, the overall differences of maternal serum UCN levels among Control, EE, IR, and EE-IR groups were significant ( = 2.48, < 0.05). Expression of WFS1 mRNA in the EE-IR group was significantly increased and higher than Control (0.46 ± 0.15 0.24 ± 0.09, < 0.05), EE (0.46 ± 0.15 0.17 ± 0.04, > 0.05), and IR (0.46 ± 0.15 0.22 ± 0.15, > 0.05) groups at 19 GD, indicating that endoplasmic reticulum stress may be activated. However, the expression of CRH (0.42 ± 0.05 0.58 ± 0.12, < 0.05), UCN (0.43 ± 0.01 0.47 ± 0.16, > 0.05), and WFS1 (0.57 ± 0.07 0.74 ± 0.12, > 0.05) protein in the EE-IR group was subsided compared to the IR group at 17 GD.
Fetal rat growth restriction was found in the EE-induced intrahepatic cholestasis model. This study revealed that significant changes in the maternal sera level of UCN , placental level of WFS1 mRNA and placental levels of CRH, UCN, and WFS1 protein in chronic versus acute stress in a rat model of pregnancy. This suggests an impaired compensatory vasodilatory effect mediated by these factors at gene transcription and protein translation levels, following acute hypoxia stress in EE-induced intrahepatic cholestasis in pregnant rats.
探讨促肾上腺皮质激素释放激素（CRH）、尿皮质素（UCN）和沃尔弗勒姆综合征1（WFS1）在17α-乙炔雌二醇（EE）诱导的肝内胆汁淤积妊娠大鼠及其缺血再灌注（IR）模型中的可能调控机制。
将60只妊娠大鼠通过随机数字表随机分为四个实验组（对照组、EE组、IR组和EE-IR组），并在妊娠第17、19和21天进行研究（每组在指定时间为5只）。记录这四组中胎鼠的生长发育指标。采用酶联免疫吸附测定法检测母鼠血清中CRH、UCN和WFS1水平。采用蛋白质印迹法和实时聚合酶链反应定量检测胎盘组织中CRH、UCN和WFS1的蛋白和mRNA水平。采用多因素方差分析和最小显著差检验进行组间和个体间比较。
在妊娠第17、19和21天，四组间胎盘重量（F = 8.10，P < 0.05）、胎鼠体重（F = 40.86，P < 0.05）、胎鼠体长（F = 61.61，P < 0.05）和胎鼠尾长（F = 55.63，P < 0.05）存在显著差异。此外，对照组、EE组、IR组和EE-IR组母鼠血清UCN水平的总体差异显著（F = 2.48，P < 0.05）。在妊娠第19天，EE-IR组中WFS1 mRNA的表达显著增加，高于对照组（0.46 ± 0.15对0.24 ± 0.09，P < 0.05）、EE组（0.46 ± 0.15对0.17 ± 0.04，P > 0.05）和IR组（0.46 ± 0.15对0.22 ± 0.15，P > 0.05），表明内质网应激可能被激活。然而，在妊娠第17天，与IR组相比，EE-IR组中CRH（0.42 ± 0.05对0.58 ± 0.12，P < 0.05）、UCN（0.43 ± 0.01对0.47 ± 0.16，P > 0.05）和WFS1（0.57 ± 0.07对0.74 ± 0.12，P > 0.05）蛋白的表达有所下降。
在EE诱导的肝内胆汁淤积模型中发现胎鼠生长受限。本研究揭示了妊娠大鼠EE诱导的肝内胆汁淤积模型中，慢性应激与急性应激相比，母鼠血清UCN水平、胎盘WFS1 mRNA水平以及胎盘CRH、UCN和WFS1蛋白水平存在显著变化。这表明在EE诱导的妊娠大鼠肝内胆汁淤积急性缺氧应激后，这些因素在基因转录和蛋白质翻译水平介导的代偿性血管舒张作用受损。
