Semaglutide May Ameliorate Fibrosis and Inhibit Epithelial-Mesenchymal Transition in Intrauterine Adhesion Models

Abstract

The purpose of this study was to explore the effect of Semaglutide on intrauterine adhesions and discover new drugs for such adhesions. In this study, the cell model was simulated by TGF-β1-induced human endometrial epithelial cells, and the animal model was established through mechanical curettage and inflammatory stimulation. After co-culturing with TGF-β1 with or without different concentrations of Semaglutide for 48 h, cells were collected for RT-qPCR and Western blotting analyses. Three doses were subcutaneously injected into experimental mice once a day for two weeks, while the control group received sterile ddH2O. The serum and uterine tissues of the mice were collected. HE and Masson staining were used for the uterine histomorphological and pathological analyses. RT-qPCR and Western blotting were used for mRNA and protein expression analyses. Serum indicators were detected using ELISA kits. The results showed that Semaglutide significantly reduced the mRNA levels of fibrosis indicators ACTA2, COL1A1, and FN and inflammatory indicators TNF-α, IL-6, and NF-κB in the two models. Semaglutide improved endometrium morphology, increased the number of endometrial glands, and reduced collagen deposition in IUA mice. The results also showed that Semaglutide could inhibit vimentin, E-Cadherin, and N-Cadherin in the two models. In summary, Semaglutide can ameliorate fibrosis and inflammation of intrauterine adhesions as well as inhibit epithelial-mesenchymal transition in IUA models.

Keywords: Semaglutide; epithelial–mesenchymal transition; fibrosis; intrauterine adhesions.

Figure 1

Viability assay of Semaglutide on HEECs induced with or without TGF-β. * p < 0.05.

Figure 2

RT-qPCR analysis of the relative mRNA expression levels of ACTA2 (A), COL1A1 (B), and FN (C) in TGF-β-induced HEECs treated with or without Semaglutide. Western blot analysis of the protein expression level of ACTA2 (D,E) in TGF-β-induced HEECs treated with or without Semaglutide. Data are expressed as mean ± SD. * p < 0.05, ** p < 0.01, and *** p < 0.001.

Figure 3

RT-qPCR analysis of the relative mRNA expression levels of TNF-α, IL-6, NF-κB, and NOX2 in TGF-β-induced HEECs treated with or without Semaglutide. Data are expressed as mean ± SD. * p < 0.05, ** p < 0.01, and *** p < 0.001.

Figure 4

Histological structures of the uterus in the six experimental groups (H&E and Masson staining). (A) Analysis of the number of glands (B) and fibrotic area (C) in the endometrium in each group at 14 days after IUA model establishment. The scale bar in the figure is 200 μΜ. Data are expressed as mean ± SD. ** p < 0.01 and *** p < 0.001.

Figure 5

RT-qPCR analysis of the relative mRNA expression levels of ACTA2 (A), COL1A1 (B), and FN (C) in the uteri of mice in each group. Western blot analysis of the protein expression level of ACTA2 in the uteri of mice in each group (D,E). Data are expressed as mean ± SD. * p < 0.05, ** p < 0.01, and *** p < 0.001.

Figure 6

RT-qPCR analysis of the relative mRNA expression levels of NOX2 (A) and NF-κB (B) in the uteri of mice in each group. ELISA analysis of the serum levels of TNF-α (C) and IL-6 (D) in mice in each group. Data are expressed as mean ± SD. * p < 0.05, ** p < 0.01, and *** p < 0.001.

Figure 7

RT-qPCR analysis of the relative mRNA expression levels of TGF-βR (A), vimentin (B), SNAIL1 (C), and N-Cadherin (D) in the TGF-β-induced HEECs in each group. Western blot analysis of the protein expression level of E-Cadherin, N-Cadherin, and vimentin in the TGF-β-induced HEECs in each group (E,F). Data are expressed as mean ± SD. * p < 0.05, ** p < 0.01, and *** p < 0.001.

Figure 8

RT-qPCR analysis of the relative mRNA expression levels of TGF-β (A), N-Cadherin (B), E-Cadherin (C), and vimentin (D) in the uteri of mice in each group. Western blot analysis of the protein expression level of TGF-β, vimentin, and TGF-β in the uteri of mice in each group (E,F). Data are expressed as mean ± SD. * p < 0.05, ** p < 0.01, and *** p < 0.001.