The Effect And Mechanism Of Semaglutide On Intrauterine Adhesion

Objective: This study intends to explore the effect and mechanism of a new longacting glucagon-like peptide(GLP-1)receptor agonist semaglutide on intrauterine adhesion(IUA)cell models and animal models,and to provide clinical Drug development for the treatment of IUA provides new ideas and theoretical basis.Method: In this study,transforming growth factor β1(TGF-β1)was used to induce endometrial stromal cells(HESC)and endometrial epithelial cells(HEEC)to establish the IUA cell model,and the method of mechanical injury combined with lipopolysaccharide stimulation was used Establish an IUA animal model.At the cellular level,the effect of drug concentration on cell viability was explored by the CCK8 method,and then qRT-PCR,WB,and Elisa were used to detect the key indicators of fibrosis α-smooth muscle actin(α-SMA),α1-1 from the mRNA and protein levels Type collagen(COL1A1),fibronectin(FN),key indicators of epithelial-mesenchymal transition(EMT)TGF-βR,snail family repressor of transcription 1(SNAIL1),vimentin(VIM),N-cadherin(N-cad),E-cadherin(E-cad)and the expression of key indicators of inflammation TNF-α,IL-6,NF-κB,NOX2,DCFH-DA method to detect the level of intracellular ROS,in order to analyze the drug effect The effect and mechanism of IUA cells.At the animal level,the number of endometrial glands and collagen fiber deposition were observed by H&E and Masson staining,and the key indicators of fibrosis and EMT in mouse uterine tissue were detected by qRT-PCR and WB α-SMA,COL1A1,FN,VIM,E-cad,TGF-β,inflammatory and oxidative stress indicators NF-κB,NOX2 were detected by qRT-PCR,inflammatory factors IL-6,TNF-α levels in mouse serum were detected by Elisa method,and semaglutide was verified Effects of peptides on animal IUA models.Result: At the cellular level,after screening by CCK8 method,semaglutide had no significant effect on the viability of HEEC and HESC at a concentration of 10 μM,and the viability of the two cells was significantly reduced at a concentration of 20 μM.mRNA expression of fibrosis markers α-SMA,FN,COL1A1,EMT markers TGF-βR,SNAIL1,VIM,N-cad,inflammation-related indicators NF-κB,TNF-α,IL-6 after TGF-β1 induction After treatment with different concentrations of semaglutide,the mRNA expressions of fibrosis markers,EMT indicators(TGF-βR was not significantly decreased),and inflammatory indicators were all decreased compared with the model group.The changes in protein expression of the above indicators were consistent with the results of mRNA expression.In addition,DCFH-DA assay showed that the expressions of ROS and NOX2 in the two cells were significantly increased after TGF-β1 treatment,but decreased after high-concentration semaglutide treatment,suggesting the presence of oxidative stress in IUA.At the animal level,H&E and Masson staining results showed that a large number of scattered glands and columnar epithelial cells were distributed on the surface of the uterine cavity of normal mice,and the number of glands and blood vessels in the uterus of mice in the model group and low-dose administration group was significantly reduced,and There was a large area of blue collagen deposition,and the number of glands and blood vessels in the uterus of mice in the middle-dose and high-dose treatment groups increased,and the area of collagen decreased significantly.The mRNA levels of fibrosis and EMT indicators α-SMA,FN,COL1A1,VIM,and TGF-βand the protein expressions of α-SMA,VIM,and TGF-β in the mice in the model group were significantly increased.All decreased,while E-cad decreased significantly in model group and low-dose administration group,and increased significantly with the increase of administration concentration.Compared with the normal group mice,the relative mRNA contents of NF-κB and NOX2,as well as the serum TNF-α and IL-6contents in the model group were significantly increased,and the above indicators were down-regulated after being treated with medium and high concentrations of semaglutide,the difference is statistically significant.Conclusion: Semaglutide can reduce the deposition of extracellular matrix in the uterine cavity,promote the proliferation of glands and blood vessels in the endometrium,and reduce the area of fibrosis,thus producing a good therapeutic effect on IUA.Preliminary mechanism research shows that this effect is related to the inhibitory effect of semaglutide on EMT,inflammation,oxidative stress and other related signaling pathways that occur during the formation of IUA.