Department of Obstetrics and Gynecology, Nanjing Maternity and Child Health Care Institute, The Affiliated Obstetrics and Gynecology Hospital with Nanjing Medical University, Nanjing, China.
Eur Rev Med Pharmacol Sci. 2019 Feb;23(3):923-931. doi: 10.26355/eurrev_201902_16978.
The aim of this study was to investigate the influence of micro-ribonucleic acid-34a (miR-34a) on preeclampsia through the Notch signaling pathway.
The expressions of miR-34a, Notch-1, Notch-2, and Notch-3 in the placenta of 39 preeclampsia patients and 42 normal patients were detected by immunohistochemistry and Reverse Transcription-Polymerase Chain Reaction (RT-PCR). The correlations between miR-34a expression with the expressions of Notch-1, Notch-2 and Notch-3 were analyzed, respectively. Besides, placental trophoblasts were isolated from preeclampsia patients and cultured in vitro. The expressions of miR-34a, Notch-1, Notch-2 and Notch-3 in placental trophoblasts were analyzed. Furthermore, the influences of miR-34a on the protein expressions of Notch-1, Notch-2, Notch-3, and hairy and enhancer of split-1 (Hes-1) in the Notch signaling pathway were analyzed by Luciferase reporter gene assay and Western blotting. The role of Notch in trophoblast invasion was investigated through the Notch inhibitors. In addition, its influence on the expression of urokinase-type plasminogen activator (uPA) was studied by miR-34a overexpression.
The expressions of miR-34a and Notch-1 were correlated with preeclampsia in the placentas of preeclampsia patients and normal patients to a certain degree. The expression of miR-34a in preeclamptic placenta was significantly higher than that of the normal placenta (p<0.05). However, Notch-1 expression was markedly lower in preeclamptic placenta (p<0.05). No significant differences were found in the expressions of Notch-2 and Notch-3 between the two types of placentas (p>0.05). MiR-34a had a remarkable negative correlation with Notch-1 expression in the Notch family (p<0.001, r=-0.5775). RT-PCR results revealed that the mRNA expression of miR-34a in placental trophoblasts of patients with preeclampsia was notably higher than that of normal people (p<0.01). However, Western blotting demonstrated that the protein expressions of Notch-1, Notch-2 and Notch-3 exhibited the opposite results. Additionally, the protein expression of Notch-1, Notch-2, Notch-3 and Hes-1 in trophoblasts transfected with pre-miR-34a was significantly decreased. The treatment with Notch inhibitors markedly reduced the trophoblast invasion. Furthermore, miR-34a overexpression or intracellular domain of Notch (ICN) overexpression regulated uPA expression.
MiR-34a regulates uPA system through the Notch signal transduction, thereby regulating the invasion of placental trophoblasts in patients with preeclampsia.
本研究旨在通过 Notch 信号通路探讨微小 RNA-34a（miR-34a）对先兆子痫的影响。
采用免疫组织化学和逆转录聚合酶链反应（RT-PCR）检测 39 例先兆子痫患者和 42 例正常患者胎盘组织中 miR-34a、Notch-1、Notch-2 和 Notch-3 的表达情况。分析 miR-34a 表达与 Notch-1、 Notch-2 和 Notch-3 表达的相关性。此外，从先兆子痫患者中分离胎盘滋养层细胞并进行体外培养。分析胎盘滋养层细胞中 miR-34a、 Notch-1、 Notch-2 和 Notch-3 的表达。此外，通过荧光素酶报告基因检测和 Western blot 分析 miR-34a 对 Notch 信号通路中 Notch-1、 Notch-2、 Notch-3 和 hairy and enhancer of split-1（Hes-1）蛋白表达的影响。通过 Notch 抑制剂研究 Notch 在滋养细胞侵袭中的作用。此外，通过 miR-34a 过表达研究其对尿激酶型纤溶酶原激活物（uPA）表达的影响。
miR-34a 和 Notch-1 的表达与先兆子痫患者胎盘组织中的先兆子痫有一定程度的相关性。先兆子痫胎盘组织中 miR-34a 的表达明显高于正常胎盘（p<0.05），而 Notch-1 的表达明显低于正常胎盘（p<0.05）。两种胎盘组织中 Notch-2 和 Notch-3 的表达无明显差异（p>0.05）。miR-34a 在 Notch 家族中与 Notch-1 表达呈显著负相关（p<0.001，r=-0.5775）。RT-PCR 结果显示，先兆子痫患者胎盘滋养层细胞中 miR-34a 的 mRNA 表达明显高于正常人（p<0.01），但 Western blot 显示 Notch-1、 Notch-2 和 Notch-3 的蛋白表达结果相反。此外，转染前 miR-34a 的滋养细胞中 Notch-1、 Notch-2、 Notch-3 和 Hes-1 的蛋白表达明显减少。Notch 抑制剂处理显著降低滋养细胞侵袭。此外，miR-34a 过表达或 Notch 细胞内结构域（ICN）过表达调节 uPA 表达。
miR-34a 通过 Notch 信号转导调节 uPA 系统，从而调节先兆子痫患者胎盘滋养层细胞的侵袭。
