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Objective: To discuss the effect of Wnt/β-catenin signaling pathway inhibitor methyl 3-｛［(4- methyl- phenyl) sulfonyl］ amino} benzoate (MSAB) on the fibrogenic response of the human endometrial stromal cells (HESCs), and to provide the foundation for the application of MSAB in the target therapy of intrauteriue adhesion (IUA). Methods: The normal HESCs were cultured in vitro and divided into two groups: control group and transforming growth factor β1 (TGF-β1) group； the HESCs from the adhesion part of the IUA patients were cultured in vitro, regarded as IUA group. Western blotting method was used to detect the expression levels of fibrotic marker protein type Ⅰ collagen α1 (COL1A1) in the cells in various groups at different time points (0, 12, 24, 48, and 60 h) after treated with TGF-β1. MTT assay was used to detect the proliferation activities of the cells in various groups. Western blotting method was used to detect the expression levels of the fibrotic marker protein COL1A1, stromal marker proteins such as N-cadherin and α-smooth muscle actin (α-SMA), and Wnt/β-catenin signaling pathway-related protein β-catenin in the cells in control and IUA groups. Based on the MSAB concentrations, the normal HESCs were divided into 0 (control), 0. 25, 0. 50, 0. 75, and 1. 00 μmol·L-1 MSAB groups, and MTT assay was used to detect the survival rates of the cells in various groups. After treated with MSAB, the normal HESCs were divided into control group (normal HESCs), TGF- β1 group (10 μg·L-1 TGF- β1 induced normal HESCs for 24 h then the drug was withdrawn, replaced with complete culture medium, and the cells continued to be cultured for 24 h), and MSAB group (10 μg·L-1 TGF-β1 induced normal HESCs for 24 h then the drug was withdrawn, replaced with a complete medium containing 0. 75 μmol·L-1 MSAB and the cells continued to be cultured for 24 h). Real-time fluorescence quantitative PCR (RT-qPCR) method was used to detect the expression levels of epithelial-mesenchymal transition (EMT) -related transcription factors Snail, Slug, Smuc, ZEB1, and ZEB2, and COL1A1 mRNA in the cells in various groups. Western blotting method was used to detect the expression levels of COL1A1, N-cadherin, α -SMA, β -catenin, and c-myc proteins in the cells in various groups. Results: Compared with control group (after treated with TGF-β1 for 0 h), the expression levels of COL1A1 proteins in the HESCs after treated with TGF-β1 for 12, 24, 48, and 60 h in TGF-β1 group were increased (P<0. 05 or P<0. 01). Compared with control group, there was no significant difference in the proliferation activity of the HESCs in IUA and TGF-β1 groups (P>0. 05). Compared with control group, the expression levels of COL1A1, β -catenin, N-cadherin, and α -SMA proteins in the cells in IUA group were increased (P<0. 05 or P< 0. 01). Compared with control group, the survival rates of the cells in 0. 75 and 1. 00 μmol·L-1 MSAB groups were decreased (P<0. 05 or P<0. 01). Compared with control group, the expression levels of Snail, Slug, and COL1A1 mRNA in the cells in TGF-β1 group were increased (P<0. 05 or P<0. 01); compared with TGF- β1 group, the expression levels of Snail, Slug, and COL1A1 mRNA in the cells in MSAB group were decreased (P<0. 05 or P<0. 01). Compared with control group, after treated with TGF-β1 for 24 h, the expression levels of COL1A1, N-cadherin, α-SMA, β -catenin, and c-myc proteins in the cells in TGF-β 1 group were increased (P<0. 01); compared with TGF-β1 group, the expression levels of COL1A1, N-cadherin, α-SMA, β-catenin, and c-myc proteins in the cells in MSAB group were decreased (P<0. 05 or P<0. 01). Conclusion: MSAB can inhibit the fibrogenic responses of the HESCs in vitro, and the results provide the theoretical basis for the application of MSAB in the target therapy of IUA. [ABSTRACT FROM AUTHOR]

OBJECTIVE: To observe the effects of Quyu Xiaomo decoction combined with warm-needling moxibustion on intrauterine adhesion, endometrial fibrosis and serum Bcl2 interaction protein 3 (BNIP3) and soluble vascular endothelial growth factor receptor-1 (sFlt-1) after medical abortion with incomplete curettage. METHODS: Totally 96 patients with medical abortion with incomplete curettage admitted into the hospital from Jan. 2020 to May 2023 were extracted to be divided into the control group and the combined group via the random number table method. All patients were given routine treatment after curettage. Forty-eight patients in the control group were given warmneedling moxibustion after curettage, and 48 patients in the combined group received Quyu Xiaomo decoction on the basis of the control group. Clinical efficacy and traditional Chinese medicine syndrome score before and after treatment were compared between two groups. The duration of abdominal pain, vaginal bleeding time and menstrual relapse time of two groups were recorded. The incidence of intrauterine adhesion was observed by hysteroscopy after treatment. Serum levels of sFIt-1, BNIP3, luteinizing hormone (LH), estrogen (E2) and tumor necrosis fact-α (TNF-α) were measured before and after treatment, mRNA expression levels of matrix metalloproteinase 9 (MMP9) and transforming growth factor β3 ( TGF-β3) in endometrial tissues were detected by reverse transcriptase polymerase chain reaction (RT-PCR). RESULTS: The total effective rate of the combined group was 97. 92% (47/48), higher than 83. 33% (40 / 48) of the control group, the difference was statistically significant (P< 0. 05). The score of lochia dripping astringency, lower abdominal pain, less dripping volume, dark purple with clumps, astringent or heavy pulse, dark purple tongue or purple spot on the edge of the postpartum in combined group was lower than that in the control group, with statistically significant differences (P<0. 05). The duration of abdominal pain, vaginal bleeding and menstrual rehydration in the combined group were shorter than those in the control group, the incidence of intrauterine adhesions in the combined group was lower than that in the control group, with statistically significant differences (P<0. 05). After treatment, the level of E2 in combined group was higher than that in control group, the levels of LH and TNF-α were lower than those in control group, BNIP3 and sFlt-1 were higher than those in control group, and MMP9 and TGF-β3 mRNA were lower than those in control group, with statistically significant differences ( P < 0. 05 ). CONCLUSIONS: Quyu Xiaomo decoction combined with warm-needling moxibustion in the treatment of patients with medical abortion with incomplete curettage can regulate sex hormone levels, improve inflammatory state, increase levels of BNIP3 and sFlt-1, improve endometrial fibrosis, enhance uterine cavity anti-adhesion, alleviate patients' disease condition and improve clinical efficacy. [ABSTRACT FROM AUTHOR]

Abstract Background Intrauterine adhesion (IUA) is one of the most severe causes of infertility in women of childbearing age with injured endometrium secondary to uterine performance. Stem cell therapy is effective in treating damaged endometrium. The current reports mainly focus on the therapeutic effects of stem cells through paracrine or transdifferentiation, respectively. This study investigates whether paracrine or transdifferentiation occurs preferentially in treating IUA. Methods Human amniotic mesenchymal stem cells (hAMSCs) and transformed human endometrial stromal cells (THESCs) induced by transforming growth factor beta (TGF-β1) were co-cultured in vitro. The mRNA and protein expression levels of Fibronectin (FN), Collagen I, Cytokeratin19 (CK19), E-cadherin (E-cad) and Vimentin were detected by Quantitative real-time polymerase chain reaction (qPCR), Western blotting (WB) and Immunohistochemical staining (IHC). The Sprague-Dawley (SD) rats were used to establish the IUA model. hAMSCs, hAMSCs-conditional medium (hAMSCs-CM), and GFP-labeled hAMSCs were injected into intrauterine, respectively. The fibrotic area of the endometrium was evaluated by Masson staining. The number of endometrium glands was detected by hematoxylin and eosin (H&E). GFP-labeled hAMSCs were traced by immunofluorescence (IF). hAMSCs, combined with PPCNg (hAMSCs/PPCNg), were injected into the vagina, which was compared with intrauterine injection. Results qPCR and WB revealed that FN and Collagen I levels in IUA-THESCs decreased significantly after co-culturing with hAMSCs. Moreover, CK19, E-cad, and Vimentin expressions in hAMSCs showed no significant difference after co-culture for 2 days. 6 days after co-culture, CK19, E-cad and Vimentin expressions in hAMSCs were significantly changed. Histological assays showed increased endometrial glands and a remarkable decrease in the fibrotic area in the hAMSCs and hAMSCs-CM groups. However, these changes were not statistically different between the two groups. In vivo, fluorescence imaging revealed that GFP-hAMSCs were localized in the endometrial stroma and gradually underwent apoptosis. The effect of hAMSCs by vaginal injection was comparable to that by intrauterine injection assessed by H&E staining, MASSON staining and IHC. Conclusions Our data demonstrated that hAMSCs promoted endometrial repair via paracrine, preferentially than transdifferentiation.

Abstract Background Endometrial fibrosis, a significant characteristic of intrauterine adhesion (IUA), is caused by the excessive differentiation and activation of endometrial stromal cells (ESCs). Glutaminolysis is the metabolic process of glutamine (Gln), which has been implicated in multiple types of organ fibrosis. So far, little is known about whether glutaminolysis plays a role in endometrial fibrosis. Methods The activation model of ESCs was constructed by TGF-β1, followed by RNA-sequencing analysis. Changes in glutaminase1 (GLS1) expression at RNA and protein levels in activated ESCs were verified experimentally. Human IUA samples were collected to verify GLS1 expression in endometrial fibrosis. GLS1 inhibitor and glutamine deprivation were applied to ESCs models to investigate the biological functions and mechanisms of glutaminolysis in ESCs activation. The IUA mice model was established to explore the effect of glutaminolysis inhibition on endometrial fibrosis. Results We found that GLS1 expression was significantly increased in activated ESCs models and fibrotic endometrium. Glutaminolysis inhibition by GLS1 inhibitor bis-2-(5-phenylacetamido-1,2,4-thiadiazol-2-yl) ethyl sulfide (BPTES or glutamine deprivation treatment suppressed the expression of two fibrotic markers, α-SMA and collagen I, as well as the mitochondrial function and mTORC1 signaling in ESCs. Furthermore, inhibition of the mTORC1 signaling pathway by rapamycin suppressed ESCs activation. In IUA mice models, BPTES treatment significantly ameliorated endometrial fibrosis and improved pregnancy outcomes. Conclusion Glutaminolysis and glutaminolysis-associated mTOR signaling play a role in the activation of ESCs and the pathogenesis of endometrial fibrosis through regulating mitochondrial function. Glutaminolysis inhibition suppresses the activation of ESCs, which might be a novel therapeutic strategy for IUA.

Background: Intrauterine adhesion (IUA) is one of the most severe causes of infertility in women of childbearing age with injured endometrium secondary to uterine performance. Stem cell therapy is effective in treating damaged endometrium. The current reports mainly focus on the therapeutic effects of stem cells through paracrine or transdifferentiation, respectively. This study investigates whether paracrine or transdifferentiation occurs preferentially in treating IUA. Methods: Human amniotic mesenchymal stem cells (hAMSCs) and transformed human endometrial stromal cells (THESCs) induced by transforming growth factor beta (TGF-β1) were co-cultured in vitro. The mRNA and protein expression levels of Fibronectin (FN), Collagen I, Cytokeratin19 (CK19), E-cadherin (E-cad) and Vimentin were detected by Quantitative real-time polymerase chain reaction (qPCR), Western blotting (WB) and Immunohistochemical staining (IHC). The Sprague-Dawley (SD) rats were used to establish the IUA model. hAMSCs, hAMSCs-conditional medium (hAMSCs-CM), and GFP-labeled hAMSCs were injected into intrauterine, respectively. The fibrotic area of the endometrium was evaluated by Masson staining. The number of endometrium glands was detected by hematoxylin and eosin (H&E). GFP-labeled hAMSCs were traced by immunofluorescence (IF). hAMSCs, combined with PPCNg (hAMSCs/PPCNg), were injected into the vagina, which was compared with intrauterine injection. Results: qPCR and WB revealed that FN and Collagen I levels in IUA-THESCs decreased significantly after co-culturing with hAMSCs. Moreover, CK19, E-cad, and Vimentin expressions in hAMSCs showed no significant difference after co-culture for 2 days. 6 days after co-culture, CK19, E-cad and Vimentin expressions in hAMSCs were significantly changed. Histological assays showed increased endometrial glands and a remarkable decrease in the fibrotic area in the hAMSCs and hAMSCs-CM groups. However, these changes were not statistically different between the two groups. In vivo, fluorescence imaging revealed that GFP-hAMSCs were localized in the endometrial stroma and gradually underwent apoptosis. The effect of hAMSCs by vaginal injection was comparable to that by intrauterine injection assessed by H&E staining, MASSON staining and IHC. Conclusions: Our data demonstrated that hAMSCs promoted endometrial repair via paracrine, preferentially than transdifferentiation. Plain English summary: IUA is the crucial cause of infertility in women of childbearing age, and no satisfactory treatment measures have been found in the clinic. hAMSCs can effectively treat intrauterine adhesions through paracrine and transdifferentiation mechanisms. This study confirmed in vitro and in vivo that amniotic mesenchymal stem cells preferentially inhibited endometrial fibrosis and promoted epithelial repair through paracrine, thus effectively treating intrauterine adhesions. The level of fibrosis marker proteins in IUA-THESCs decreased significantly after co-culturing with hAMSCs for 2 days in vitro. However, the level of epithelial marker proteins in hAMSCs increased significantly, requiring at least 6 days of co-culture. hAMSCs-CM had the same efficacy as hAMSCs in inhibiting fibrosis and promoting endometrial repair in IUA rats, supporting the idea that hAMSCs promoted endometrial remodeling through paracrine in vivo. In addition, GFP-labeled hAMSCs continuously colonized the endometrial stroma instead of the epithelium and gradually underwent apoptosis. These findings prove that hAMSCs ameliorate endometrial fibrosis of IUA via paracrine, preferentially than transdifferentiation, providing the latest insights into the precision treatment of IUA with hAMSCs and a theoretical basis for promoting the "cell-free therapy" of MSCs. [ABSTRACT FROM AUTHOR]

Background: Endometrial fibrosis, a significant characteristic of intrauterine adhesion (IUA), is caused by the excessive differentiation and activation of endometrial stromal cells (ESCs). Glutaminolysis is the metabolic process of glutamine (Gln), which has been implicated in multiple types of organ fibrosis. So far, little is known about whether glutaminolysis plays a role in endometrial fibrosis. Methods: The activation model of ESCs was constructed by TGF-β1, followed by RNA-sequencing analysis. Changes in glutaminase1 (GLS1) expression at RNA and protein levels in activated ESCs were verified experimentally. Human IUA samples were collected to verify GLS1 expression in endometrial fibrosis. GLS1 inhibitor and glutamine deprivation were applied to ESCs models to investigate the biological functions and mechanisms of glutaminolysis in ESCs activation. The IUA mice model was established to explore the effect of glutaminolysis inhibition on endometrial fibrosis. Results: We found that GLS1 expression was significantly increased in activated ESCs models and fibrotic endometrium. Glutaminolysis inhibition by GLS1 inhibitor bis-2-(5-phenylacetamido-1,2,4-thiadiazol-2-yl) ethyl sulfide (BPTES or glutamine deprivation treatment suppressed the expression of two fibrotic markers, α-SMA and collagen I, as well as the mitochondrial function and mTORC1 signaling in ESCs. Furthermore, inhibition of the mTORC1 signaling pathway by rapamycin suppressed ESCs activation. In IUA mice models, BPTES treatment significantly ameliorated endometrial fibrosis and improved pregnancy outcomes. Conclusion: Glutaminolysis and glutaminolysis-associated mTOR signaling play a role in the activation of ESCs and the pathogenesis of endometrial fibrosis through regulating mitochondrial function. Glutaminolysis inhibition suppresses the activation of ESCs, which might be a novel therapeutic strategy for IUA. [ABSTRACT FROM AUTHOR]

Abstract Macrophages are a key and heterogeneous cell population involved in endometrial repair and regeneration during the menstrual cycle, but their role in the development of intrauterine adhesion (IUA) and sequential endometrial fibrosis remains unclear. Here, we reported that CD301+ macrophages were significantly increased and showed their most active interaction with profibrotic cells in the endometria of IUA patients compared with the normal endometria by single‐cell RNA sequencing, bulk RNA sequencing, and experimental verification. Increasing CD301+ macrophages promoted the differentiation of endometrial stromal cells into myofibroblasts and resulted in extracellular matrix accumulation, which destroyed the physiological architecture of endometrial tissue, drove endometrial fibrosis, and ultimately led to female infertility or adverse pregnancy outcomes. Mechanistically, CD301+ macrophages secreted GAS6 to activate the AXL/NF‐κB pathway, upregulating the profibrotic protein synthesis. Targeted deletion of CD301+ macrophages or inhibition of AXL by Bemcentinib blunted the pathology and improved the outcomes of pregnancy in mice, supporting the therapeutic potential of targeting CD301+ macrophages for treating endometrial fibrosis.

Macrophages are a key and heterogeneous cell population involved in endometrial repair and regeneration during the menstrual cycle, but their role in the development of intrauterine adhesion (IUA) and sequential endometrial fibrosis remains unclear. Here, we reported that CD301+ macrophages were significantly increased and showed their most active interaction with profibrotic cells in the endometria of IUA patients compared with the normal endometria by single‐cell RNA sequencing, bulk RNA sequencing, and experimental verification. Increasing CD301+ macrophages promoted the differentiation of endometrial stromal cells into myofibroblasts and resulted in extracellular matrix accumulation, which destroyed the physiological architecture of endometrial tissue, drove endometrial fibrosis, and ultimately led to female infertility or adverse pregnancy outcomes. Mechanistically, CD301+ macrophages secreted GAS6 to activate the AXL/NF‐κB pathway, upregulating the profibrotic protein synthesis. Targeted deletion of CD301+ macrophages or inhibition of AXL by Bemcentinib blunted the pathology and improved the outcomes of pregnancy in mice, supporting the therapeutic potential of targeting CD301+ macrophages for treating endometrial fibrosis. Synopsis: This study highlights the role of CD301+ macrophages in facilitating endometrial fibrosis in intrauterine adhesion (IUA) through the GAS6/AXL/NF‐κB pathway. By depleting CD301+ macrophages or employing the pharmacological inhibitor Bemcentinib to target AXL, the progression of fibrosis can be suppressed, ultimately improving pregnancy outcomes in mice. CD301+ macrophages exhibit elevated levels in the endometrial fibrosis of IUA patients.CD301+ macrophages secrete GAS6, activating the AXL/NF‐κB pathway and promoting fibrosis.Depletion of CD301+ macrophages or treatment with the AXL inhibitor Bemcentinib shows promising potential for treating endometrial fibrosis and enhancing pregnancy outcomes. [ABSTRACT FROM AUTHOR]

Intrauterine adhesion (IUA) is a common gynecological disease with limited therapeutic options. Dulaglutide is a long-acting glucagon-like peptide-1 (GLP-1) analog with some anti-fibrotic and anti-inflammatory properties; however, its action on IUA remains uncertain. The purpose of the experiments in this study was to explore the effect of dulaglutide on IUA and to elucidate its mechanism to provide new ideas for the clinical treatment of IUA. An IUA mouse model was established via mechanical curettage and inflammation induction; mice received subcutaneous injection with three doses of dulaglutide once a day for two weeks (treatment) or equal amounts of sterile ddH2O (control), and sham-operated mice were treated similarly to the control mice. Mice were sacrificed, and uterine tissues were subjected to hematoxylin and eosin (H&E) and Masson's trichrome staining for histomorphological and pathological analyses and real-time quantitative polymerase chain reaction (RT-qPCR) and Western blotting (WB) for gene and protein expression analyses. Dulaglutide improved the shape of the uterine cavity, increased endometrial thickness and the number of glands, and significantly reduced the area of collagen fiber deposition in the endometrium. It significantly reduced collagen type I A 1 (COL1A1), interleukin-1beta (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α), C-C motif chemokine ligand 2 (CCL2), F4/80 (macrophage), vimentin and transforming growth factor-beta (TGF-β) mRNA levels and COL1A1, IL-1β, IL-6, TNF-α, F4/80, vimentin, E-cadherin, TGF-β, and p-Smad2 protein expression levels. This study demonstrates that dulaglutide reduces inflammatory responses by inhibiting M1 macrophage polarization and inflammatory factor release and may ameliorate fibrosis by inhibiting epithelial–mesenchymal transition (EMT) via TGF-β/Smad2 signaling. [ABSTRACT FROM AUTHOR]

Objective To investigate the effects of Yuzhi Zhixue granules on stromal cell-derived factor-1 (SDF-1)/CXC chemokine receptor 4 (CXCR4) axis and endometrial fibrosis in rats with intrauterine adhesions. Methods The rats were randomly divided into sham operation group, model group and Yuzhi Zhixue granules low, medium and high dose groups(to establish a rat model of intrauterine adhesions), and were given 30 mg/kg, 60 mg/kg and 120 mg/kg by gavage respectively, once a day for 21 consecutive days. The levels of serum interleukin-6(IL-6), tumor necrosis factor-α (TNF-α) and transforming growth factor-β1 (TGF-β1) were detected by kits; HE staining and Masson staining were used to detect the number of endometrial glands and the degree of interstitial fibrosis; the protein expressions of SDF-1 and CXCR4 were detected by Western blot. Results Compared with those in the sham operation group, the uterine tissue structure of the model group was destroyed, the number of glands decreased, fibrotic hyperplasia was found and a large number of inflammatory cells infiltration was found. The levels of serum IL-6, TNF-α and TGF-β1 were significantly higher (P＜0.05) and the expression of SDF-1 protein in uterine tissue was significantly lower (P＜0.05). Compared with those in the model group, the number of glands in the low, medium and high dose groups of Yuzhi Zhixue Granules increased in turn, the degree of fibrosis was weakened, and the number of inflammatory cells was decreased, the levels of IL-6, TNF-α and TGF-β1 in serum were significantly lower (P＜0.05). The expression of SDF-1 protein in uterus was significantly lower (P＜0.05), and all of them were dose-dependent. Conclusions Yuzhi Zhixue granules can repair the uterine structure of rats with intrauterine adhesions, alleviate tissue fibrosis and reduce inflammatory reaction by inhibition of SDF-1/CXCR4 axis.

Abstract Background Endometrial fibrosis caused by intrauterine adhesion (IUA) can lead to hypomenorrhea, amenorrhea, and even infertility and abortion. The postoperative recurrence rate of severe IUA remains high, giving rise to low pregnancy rates. An extracellular matrix (ECM) scaffold, a new biological material that can promote cell proliferation and differentiation at lesions, has been widely used in general surgery and neurosurgery. The present study applied ECM scaffolds in obstetrics and gynecology for the first time to improve endometrial fibrosis, repair severe IUA, and improve pregnancy outcomes for infertile patients. Methods This paper presents a prospective randomized single-blind controlled superiority study of infertile women aged ≤40 years with IUA. According to the scoring criteria for IUA established by the American Fertility Society, patients with moderate or severe IUA were randomized into two groups at a ratio of 1:1; patients in the experimental group were treated with an ECM scaffold (small intestinal submucosa [SIS]) + intrauterine balloon, while patients in the control group were treated with an intrauterine balloon only. A hysteroscopic examination of adhesion repair was performed again after 2 months of postoperative hormone replacement therapy. Endometrial tissue was sampled during the two operations, and immunohistochemistry was used to observe endometrial and microvascular proliferation. After thawing and resuscitation, a postoperative frozen embryo transfer was performed on the participants in both groups, and their endometrial thickness, intrauterine volume, endometrial vascularization flow index, endometrial flow index, and uterine artery blood flow resistance were evaluated by 3D ultrasonography. The rates of embryo implantation, clinical pregnancy, and early spontaneous abortion were observed. Discussion The ECM scaffold (SIS) + intrauterine balloon method was able to repair endometrial fibrosis and improve IUA. This new technique represents a novel treatment method for improving the pregnancy outcome of infertile patients with moderate/severe IUA. Trial registration Chinese Clinical Trial Register ChiCTR2100052027 . Registered on October 14, 2021.

Intrauterine adhesion (IUA) is a common gynecological disease with limited therapeutic options. Dulaglutide is a long-acting glucagon-like peptide-1 (GLP-1) analog with some anti-fibrotic and anti-inflammatory properties; however, its action on IUA remains uncertain. The purpose of the experiments in this study was to explore the effect of dulaglutide on IUA and to elucidate its mechanism to provide new ideas for the clinical treatment of IUA. An IUA mouse model was established via mechanical curettage and inflammation induction; mice received subcutaneous injection with three doses of dulaglutide once a day for two weeks (treatment) or equal amounts of sterile ddH2O (control), and sham-operated mice were treated similarly to the control mice. Mice were sacrificed, and uterine tissues were subjected to hematoxylin and eosin (H&E) and Masson’s trichrome staining for histomorphological and pathological analyses and real-time quantitative polymerase chain reaction (RT-qPCR) and Western blotting (WB) for gene and protein expression analyses. Dulaglutide improved the shape of the uterine cavity, increased endometrial thickness and the number of glands, and significantly reduced the area of collagen fiber deposition in the endometrium. It significantly reduced collagen type I A 1 (COL1A1), interleukin-1beta (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α), C-C motif chemokine ligand 2 (CCL2), F4/80 (macrophage), vimentin and transforming growth factor-beta (TGF-β) mRNA levels and COL1A1, IL-1β, IL-6, TNF-α, F4/80, vimentin, E-cadherin, TGF-β, and p-Smad2 protein expression levels. This study demonstrates that dulaglutide reduces inflammatory responses by inhibiting M1 macrophage polarization and inflammatory factor release and may ameliorate fibrosis by inhibiting epithelial–mesenchymal transition (EMT) via TGF-β/Smad2 signaling.

Objective The intrauterine adhesion（IUA）model was established by mechanical curettage after em⁃ bryo removal by cesarean section in early-trimester rats，and its function was evaluated. Methods Forty-eight SD female rats with stable estrous cycles aged 8-9 weeks were randomly divided into groups A，B，C，and D，with 12 rats in each group. In the groups A，B，and C，after female mice and male mice were placed together，we screened pregnant rats（re⁃ corded as the 0. 5th day of pregnancy）. On the 5. 5th to 6. 5th day of pregnancy，the IUA model was established by me⁃ chanical curettage after the embryos were removed by cesarean section in the first trimester. The female rats in the group D were raised alone without male rats. We sutured and closed their abdomen without curettage. Rats in the groups A，B，and C were continued to feed for 1，2，and 3 estrous cycles after curettage. Six rats in each group were sacrificed，and uterine tissues were collected. After HE staining，the number of endometrial glands in each group was calculated. The degree of endometrial fibrosis in rats of each group was observed by using Masson staining method，and TGF-β1 in the endometrial tissues was detected by immunohistochemistry. The remaining six rats in each group were randomly assigned to male rats and caged，and the pregnant rats were sacrificed on the 14. 5th day of gestation，and the number of embryos in the uterus was recorded. Results Compared with group D，rats in groups A，B，and C had fewer endometrial glands，higher scores of endometrial fibrosis，and higher relative expression of TGF-β1 （all P<0. 05）. Compared with groups B and C，the num⁃ ber of endometrial glands was more，the score of fibrosis was lower，and the relative expression of TGF-β1 was lower in the group A（all P<0. 05）. On the 14. 5th day of pregnancy，the number of bilateral uterine embryos in the groups A，B，C，and D were 2. 08±1. 16，0，0，and 6. 75±1. 14，respectively. Compared with group D，the number of bilateral uterine em⁃ bryos in the group A was smaller（P<0. 05）. Conclusions The IUA rat model is successfully established by mechanical curettage after abortion in early-trimester rats. The model rats have decreased pregnancy ability，decreased number of en⁃ dometrial glands，increased fibrosis，and increased TGF-β1 expression. [ABSTRACT FROM AUTHOR]

Background: Endometrial fibrosis caused by intrauterine adhesion (IUA) can lead to hypomenorrhea, amenorrhea, and even infertility and abortion. The postoperative recurrence rate of severe IUA remains high, giving rise to low pregnancy rates. An extracellular matrix (ECM) scaffold, a new biological material that can promote cell proliferation and differentiation at lesions, has been widely used in general surgery and neurosurgery. The present study applied ECM scaffolds in obstetrics and gynecology for the first time to improve endometrial fibrosis, repair severe IUA, and improve pregnancy outcomes for infertile patients.Methods: This paper presents a prospective randomized single-blind controlled superiority study of infertile women aged ≤40 years with IUA. According to the scoring criteria for IUA established by the American Fertility Society, patients with moderate or severe IUA were randomized into two groups at a ratio of 1:1; patients in the experimental group were treated with an ECM scaffold (small intestinal submucosa [SIS]) + intrauterine balloon, while patients in the control group were treated with an intrauterine balloon only. A hysteroscopic examination of adhesion repair was performed again after 2 months of postoperative hormone replacement therapy. Endometrial tissue was sampled during the two operations, and immunohistochemistry was used to observe endometrial and microvascular proliferation. After thawing and resuscitation, a postoperative frozen embryo transfer was performed on the participants in both groups, and their endometrial thickness, intrauterine volume, endometrial vascularization flow index, endometrial flow index, and uterine artery blood flow resistance were evaluated by 3D ultrasonography. The rates of embryo implantation, clinical pregnancy, and early spontaneous abortion were observed.Discussion: The ECM scaffold (SIS) + intrauterine balloon method was able to repair endometrial fibrosis and improve IUA. This new technique represents a novel treatment method for improving the pregnancy outcome of infertile patients with moderate/severe IUA.Trial Registration: Chinese Clinical Trial Register ChiCTR2100052027 . Registered on October 14, 2021. [ABSTRACT FROM AUTHOR]

Objective: Intrauterine adhesion (IUA) is a major health problem that causes infertility, menstrual irregularities, and recurrent pregnancy losses in women. Unfortunately, treatments for IUA are limited, and there are currently no effective strategies for preventing IUA recurrence. In this review, we introduced the role of Hippo signaling in the normal endometrium and IUA and described the mechanisms by which the Hippo pathway integrates with the Wnt and transforming growth factor-β (TGF-β) signaling pathways to form an intricate network governing the development of fibrosis. Data Sources: Original research articles in English that were published until July 2017 were collected from the PubMed database. Study Selection: Literature search was conducted using the search terms “endometrial fibrosis OR fibrosis AND or OR intrauterine adhesion OR Asherman syndrome OR IUA,” “Hippo AND or OR Hippo/TAZ,” “TGF-β,” and “Wnt.” Related original research articles were included in the comprehensive analysis. Results: Endometrial fibrosis is recognized as a key pathological event in the development of IUA, which is characterized by epithelial/fibroblast–myofibroblast transition. Myofibroblasts play crucial roles in the pathogenesis of fibrous scarring, and myofibroblast differentiation can be triggered by multiple signaling pathways. Hippo signaling is a critical regulator of the epithelial/fibroblast–myofibroblast transition and α-smooth muscle actin, which exhibits a specific spatiotemporal expression in the endometrium. Conclusions: Hippo signaling plays a critical role in fibrous diseases and participates in cross talks with Wnt and TGF-β signaling. Our findings not only contributed to knowledge on the pathogenesis of endometrial fibrosis, but can also serve as a useful resource for developing specific molecular inhibitors for IUA treatment and prevention.

Intrauterine adhesion (IUA), characterized by endometrial fibrosis, may lead to infertility and recurrent pregnancy loss. At present, there is no ideal therapy for IUA. Recent findings have revealed that microRNAs (miRNAs) have a decisive role in the regulation of fibrosis. The aim of the present study was to investigate the molecular mechanism of miRNAs in endometrial fibrosis. The present study compared the expression profiles of miRNAs between endometrial tissues from patients with IUA and normal endometrial tissues using microarray analysis. Validation of miR‑326 level in endometrial tissues was performed using reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR). Subsequently, the effects of miR‑326 on fibrotic markers including α‑smooth muscle actin (α‑SMA), collagen type I α 1 chain (COL1A1), transforming growth factor‑β1 (TGF‑β1) and fibronectin (FN), were evaluated in endometrial tissues and endometrial stromal cells (ESCs) from patients with IUA. Additional bioinformatics analysis, luciferase reporter assays, RT‑qPCR and western blotting were performed to identify target genes. Additionally, the expression levels of TGF‑β1, p‑Smad3 and Smad3 were quantified to determine whether the anti‑fibrotic role of miR‑326 was associated with the activity of the TGF‑β1/Smad3 signaling pathway. The present study determined that miR‑326 was downregulated in endometrial tissues from patients with IUA and miR‑326 levels were inversely correlated with the expression of TGF‑β1, α‑SMA, COL1A1 and FN. Additional findings revealed that overexpression of miR‑326 inhibited endometrial fibrosis by downregulating these pro‑fibrotic genes. TGF‑β1, an important pro‑fibrogenic mediator, was identified as a direct target of miR‑326. Additionally, overexpression of miR‑326 blocked the activation of the TGF‑β1/SMAD family member 3 (Smad3) signaling pathway by suppressing the expression of TGF‑β1 in ESCs from patients with IUA. The findings of the present study indicated that miR‑326 inhibited endometrial fibrosis by suppressing the TGF‑β1/Smad3 signaling pathway, suggesting that miR‑326 may be a prognostic biomarker and therapeutic target for IUA. [ABSTRACT FROM AUTHOR]