妊娠期糖尿病患者胎盘组织中miRNA-508-3p和HGF表达水平及其对滋养细胞胰岛素抵抗的影响

doi:10.13481/j.1671-587x.20210126

Abstract

Abstract: Objective

To investigate the expressions of microRNA-508-3p （miR-508-3p） and hepatocyte growth factor （HGF） in placenta tissue of the patients with gestational diabetes mellitus （GDM） and their relationship， and to clarify the related molecular mechanism of miR-508-3p in mediating trophoblast insulin resistance（IR）.

Methods

A total of 15 GDM patients and 15 normal pregnant women were selected and used as GDM group and normal control group. The expression levels of miR-508-3p and HGF protein in placenta tissue of the patients in two groups were detected by RT-PCR， immunohistochemistry and Western blotting methods， and their relationship was analyzed.HGF was screened as the target gene of miR-508-3p with bioinformatics，which was verified by double luciferase reporter gene assay and Western blotting method. The human trophoblast cells HTR-8/Svneo were cultured in vitro to establish the HTR-8/Svneo IR cell model （HTR-8/Svneo-IR）. The HTR-8/Svneo-IR cells were transfected with miR-508-3p-inhibitor （miR-508-3p-inhibitor-IR grouop） and miR-NC（miR-NC-IR group） by liposome transient transfection method. The transfection efficiency was detected by RT-PCR； Western blotting method was used to detect the expression levels of HGF and phosphorylated phosphatidylinosital 3 kinase（p-PI3K） and phosphorylated protein kinse B（p-AKT） proteins in PI3K/AKT signaling pathway in the cells in various groups.

Results

Compared with normal control group， the expression levels of miR-508-3p in placenta tissue of the patients in GDM group was significantly increased （P<0.05）， while the HGF protein expression level was significantly decreased （P<0.01）， and there was a negative correlation between the expression level of HGF protein and the expression level of miR-508-3p in placenta tissue of the patients in GDM group（r =－0.542， P<0.05）. Compared with HTR-8/Svneo group， the expression level of miR-508-3p in the cells in HTR-8/Svneo-IR group was significantly increased （P<0.05）；compared with HTR-8/Svneo-IR group，the expression level of miR-508-3p in the cells in miR-508-3p-inhibitor-IR group was significantly decreased （P<0.05）.Compared with HTR-8/Svneo group， the expression levels of HGF，p-PI3K and p-AKT proteins in the cells in HTR-8/Svneo-IR group were significantly decreased （P<0.05）；compared with HTR-8/Svneo-IR group， the expression levels of HGF，p-PI3K and p-AKT proteins in the cells in miR-508-3p-inhibitor-IR group were significantly increased （P<0.05）.

Conclusion

MiR-508-3p can inhibit the expression of HGF in placenta tissue of the GDM patients， and promote IR of trophoblasts through PI3K/AKT signaling pathway， and then participate in the occurrence and development of GDM.

Key words: gestational diabetes mellitus, microRNA-508-3p, hepatocyte growth factor, insulin resistance, trophoblasts

CLC Number:

R714. 256

Hao HUANG,Hong JIA,Xiaoshuang WANG,Lu ZHANG,Yating DUAN. Expressions of miRNA-508-3p and HGF in placenta tissue of patients with gestational diabetes mellitus and their effects on trophoblast insulin resistance[J].Journal of Jilin University(Medicine Edition), 2021, 47(1): 187-195.

Figures/Tables 8

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References 23

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Group	HGF	p-PI3K	p-AKT
HTR-8/Svneo	0.54±0.11	0.34±0.09	0.13±0.01
HTR-8/Svneo-IR	0.21±0.02^*	0.11±0.02^*	0.06±0.02^*
miR-NC-IR	0.19±0.05^*	0.09±0.01^*	0.04±0.01^*
miR-508-3p-inhibitor-IR	0.81±0.07^?	0.77±0.10^?	0.51±0.04^?

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Expressions of miRNA-508-3p and HGF in placenta tissue of patients with gestational diabetes mellitus and their effects on trophoblast insulin resistance

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