miR-138-5p protects β cell function by regulating HIF-1α in gestational diabetes mellitus
10.12092/j.issn.1009-2501.2020.05.002
- VernacularTitle: 妊娠期糖尿病中患者miR-138-5p通过调控缺氧诱导因子1α保护β细胞功能
- Author:
Hao HUANG
1
Author Information
1. Department of Gynecology and Obstetrics, Chongqing Traditional Chinese Medicine Hospital
- Publication Type:Journal Article
- Keywords:
Gestational diabetes mellitus;
HIF-1 α;
INS-1 cells;
MiR-138-5p;
PI3K/Akt signaling pathway
- From:
Chinese Journal of Clinical Pharmacology and Therapeutics
2020;25(5):489-497
- CountryChina
- Language:Chinese
-
Abstract:
AIM: To study the effect of miR-138-5p on the function of β cell in gestational diabetes mellitus (GDM) and its related mechanism. METHODS: The expression of miR-138-5p in peripheral blood of 15 GDM pregnant women and 15 normal pregnant women were compared by RT-qPCR. miR-138-5p mimic and inhibitor were transfected into INS-1 cells, respectively, and their expression level was over expressed or inhibited. RT-qPCR was used to verify the transfection efficiency.MTT proliferation experiment, Annexin V-FITC apoptosis experiment and insulin release experiment were used to detect the effects of miR-138-5p on INS-1 cell proliferation, apoptosis and insulin release ability. The target gene of miR-138-5p was screened by TargetScan, a miRNA target gene prediction software. The functional rescue experiment confirmed whether miR-138-5p could exert its influence on INS-1 cell proliferation, apoptosis and insulin release ability by targeting its target gene. Western blot was used to detect the molecular signaling pathway of miR-138-5p in INS-1 cells.RESULTS: The expression of miR-138-5p in peripheral blood of GDM pregnant women was significantly lower than that of normal pregnant women. RT-qPCR showed that miR-138-5p mimic and inhibitor could significantly promote or inhibit the expression of miR-138-5p in INS-1 cells. The results of MTT proliferation experiment, Annexin V-FITC apoptosis experiment and insulin release experiment indicated that over expression of miR-138-5p could significantly promote the proliferation of INS-1 cells, inhibit the apoptosis of cells and promote the insulin release ability of cells. However, down-regulating the expression of miR-138-5p could significantly inhibit the proliferation of INS-1 cells, promote apoptosis and inhibit insulin release. HIF-1α was selected as the target gene of miR-138-5p by TargetScan. The double luciferase gene report and Western blot showed that miR-138-5p could inhibit the expression of HIF-1α in INS-1 cells. The functional rescue experiment confirmed that miR-138-5p could affect the proliferation, apoptosis and insulin release of INS-1 cells by regulating the expression of HIF-1α. Western blot showed that miR-138-5p may play a role in INS-1 cells by affecting PI3K, Akt and p-PI3K, p-Akt protein after phosphorylation in PI3K/Akt signaling pathway. CONCLUSION: miR-138-5p may reguLate HIF-1α expression in a targeted manner, thereby affecting the PI3K/AKT signaling pathway, promoting the proliferation and inhibition of the parent cells' apoptosis, and promoting their insulin-releasing ability to protect the function of β cell in GDM.
