IL-37 Ameliorates Chronic Endometritis by Attenuating Epithelial-Mesenchymal Transition and Promoting M2 Macrophage Polarization.

The pET22b-TAT-IL37d plasmid was kindly provided by Shandong University, and was used for expressing the fusion protein of the TAT transmembrane peptide and human IL-37d (TAT-IL-37d), thereby enabling efficient intracellular delivery of exogenous IL-37d [ 27 ]. For protein expression, the plasmid was introduced into E. coli BL21(DE3), a T7 expression host compatible with the pET22b vector. Cultures were grown from glycerol stocks in LB medium supplemented with ampicillin. After the log phase was reached, protein expression was induced with IPTG at 16 °C. Cells were lysed by ultrasonication, and the supernatant was purified through Ni-NTA affinity chromatography. After dialysis, endotoxin removal, and testing, the samples were confirmed to meet the required standards. The protein concentration was measured using a BCA assay, and protein purity was assessed through SDS-PAGE. Finally, the protein was filter-sterilized, aliquoted, and stored at −80 °C. All protein batches used for cell and animal experiments were quantitatively tested using the Limulus Amebocyte Lysate (LAL) method, and the concentration used was significantly lower than the permissible limit for routine cell experiments (typically 0.1 EU/mL). NRK-52e, HEK-293T, and RAW 264.7 cells were all purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). All cells were cultured in high-glucose DMEM (Gibco, Grand Island, NY, USA) supplemented with 10% fetal bovine serum (Gibco) and 1% penicillin-streptomycin (Gibco) and maintained at 37 °C in an incubator under 5% CO 2 . All animal protocols were approved by the Biomedical Research Ethics Committee of Hunan Normal University (permit number: 2025674; approval date: 1 September 2025). This study strictly adhered to the Animal Research ARRIVE reporting guidelines. Eight-week-old female Sprague-Dawley rats ( n = 35) were housed in an SPF-grade facility and randomly divided into four groups: control ( n = 8), LPS ( n = 9), 1 μg of IL-37 ( n = 9), and 2 μg of IL-37 ( n = 9). The sample size was determined based on previous similar studies and our preliminary experiments to ensure adequate statistical power. The two IL-37 doses were selected based on the established effective range reported in rat models of inflammatory diseases [ 28 , 29 , 30 ]. CE was induced by uterine curettage combined with LPS suture embedding. The control group underwent laparotomy and exposure of the uterine cornua only, without curettage or embedding. After the suture was removed, the treatment groups received intraperitoneal injections of IL-37 every 3 days. The control and LPS groups were administered the same volume of saline. The rats were euthanized at the third estrous cycle after model establishment. Uterine tissues were then collected for examination. The paraffin sections were deparaffinized, hydrated, stained with hematoxylin for 5 to 8 min and rinsed in running water. They were then counterstained with eosin for 1 to 3 min, dehydrated in a graded ethanol series, cleared in xylene, and mounted with neutral balsam. The sections were then observed under a microscope, and the images were analyzed using ImageJ (version 1.54f) software. Following dewaxing and hydration, the paraffin sections were stained with Weigert’s iron hematoxylin for 5–10 min, differentiated with acid ethanol, and subsequently blued. They were then immersed in Lichun red and acid eosin for 5–10 min and treated with phosphomolybdic acid for 5 min. Without washing in water, the sections were counterstained with aniline blue for 5 min, rinsed with a weak acid solution, dehydrated in a graded ethanol series, cleared with xylene, and mounted with neutral gum. The samples were examined microscopically, photographed, and analyzed using ImageJ software. After being embedded in OCT, frozen tissue sections were prepared, fixed, permeabilized, and blocked. The sections were then incubated overnight at 4 °C with primary antibodies against the following proteins: IL-37 (Abcam, Cambridge, UK), E-cadherin (Cell Signaling Technology [CST], Danvers, MA, USA), vimentin (ABclonal, Woburn, MA, USA), CD86 (Proteintech, Rosemont, IL, USA), CD206 (Proteintech), Smad3 (CST), and STAT6 (Proteintech). The following day, the sections were incubated with fluorescent secondary antibodies at room temperature in the dark for 1 h, and then stained with DAPI. The sections were imaged through microscopy and analyzed using ImageJ. Transwell chambers were placed in a 24-well plate. A cell suspension in serum-free medium was added to the upper chamber, and complete medium supplemented with 10% fetal bovine serum was added to the lower chamber as a chemotactic agent. The plates were incubated at 37 °C under 5% CO 2 for 24 h. After incubation, the medium was removed from the upper chamber, and nonmigrated cells were wiped from the upper surface with a cotton swab. The migrated cells on the lower surface were fixed with 4% paraformaldehyde, stained with 0.1% crystal violet, and analyzed using ImageJ. Total protein was extracted using RIPA lysis buffer. Equal amounts of protein were separated by 10% SDS-PAGE and transferred to PVDF membranes. After being blocked with 5% nonfat milk, the membranes were incubated overnight at 4 °C with primary antibodies against E-cadherin (CST), vimentin (ABclonal), STAT6 (Proteintech), p-STAT6 (CST), Smad3 (CST), p-Smad3 (CST), Arg1 (CST), and β-actin (CST). The next day, the membranes were incubated with the secondary antibody for 1 h at room temperature. Chemiluminescence was detected, and images were captured using a Tanon system, followed by grayscale analysis using ImageJ. Total RNA was extracted using TRIzol (Invitrogen, Carlsbad, CA, USA). The concentration and purity were measured, and cDNA was subsequently synthesized with a reverse transcription kit (Takara Bio, Kusatsu, Japan). qPCR was performed with a TAKARA kit on a Bio-Rad instrument using the following conditions: 95 °C for 30 s, followed by 40 cycles of 95 °C for 5 s and 60 °C for 30 s. The primer sequences used for qPCR are listed in Appendix A   Table A1 . β-actin or GAPDH served as internal controls, and relative gene expression was determined by the 2 −ΔΔCt method. RAW 264.7 cells from each group were collected and incubated with Fc blocking reagent (BD Biosciences, Franklin Lakes, NJ, USA) on ice for 15 min to prevent nonspecific binding. Afterward, the cells were fixed, permeabilized and incubated with an APC-conjugated anti-CD206 antibody (BioLegend, San Diego, CA, USA) in the dark at room temperature for 30 min. The cells were subsequently washed twice with precooled PBS and detected using a BD flow cytometer. HEK-293T cells were cotransfected with the pLenti-Smad-minP-Luc reporter plasmid and the pRL-TK internal reference plasmid using Lipo8000TM transfection reagent (Beyotime, Shanghai, China). After 24 h, the cells were treated with 10, 100, or 1000 ng/mL Il-37 for an additional 24 h. The cells were then lysed, and luciferase activity was measured using a dual-luciferase reporter assay kit (Beyotime). Smad pathway transcriptional activity is expressed as the ratio of firefly luciferase activity to Renilla luciferase activity. All the data were derived from at least three independent experiments and are expressed as the mean ± standard deviation. Statistical analysis was performed using GraphPad Prism 8.0 software. Unpaired t tests were employed for comparisons between two groups, one-way ANOVA for multigroup comparisons, and Tukey’s post hoc test for pairwise comparisons between groups. A p value < 0.05 was considered to indicate statistical significance.

Chronic endometritis (CE), a persistent inflammatory condition of the endometrium characterized by stromal plasma cell infiltration, edema, and aberrant glandular architecture, significantly contributes to infertility, recurrent implantation failure, and miscarriage [ 1 , 2 ]. Endometrial epithelial and stromal cells recognize pathogen-associated molecular patterns (PAMPs) or damage-associated molecular patterns (DAMPs) and activate the Toll-like receptor signaling pathway, thereby initiating NF-κB and MAPK cascade reactions. This activation promotes the sustained release of inflammatory cytokines and chemokines, perpetuating immune cell infiltration [ 3 , 4 ]. The resulting chronic inflammatory milieu disrupts glandular function and endometrial receptivity, leading to progressive tissue fibrosis characterized by excessive collagen deposition, endometrial sclerosis, and loss of plasticity [ 5 , 6 ]. Current management strategies, predominantly antibiotic-based strategies, face significant challenges, including microbial resistance, disease recurrence, and the inability to reverse established fibrosis, thereby underscoring the urgent need for novel therapeutic strategies that simultaneously target inflammation and fibrotic remodeling [ 7 , 8 , 9 ]. Interleukin-37 (IL-37) is a multifunctional cytokine with potent immunosuppressive activity within the IL-1 family. Intracellular IL-37 binds to Smad3 and is translocated to the nucleus, leading to the upregulation of PTPN expression. This upregulation inhibits the activation of key proinflammatory signaling pathways, including ERK, p38, JNK, PI3K, and NF-κB [ 10 , 11 , 12 , 13 , 14 ]. Extracellularly, IL-37 binds to the IL-18Rα/IL-1R8 complex, which activates inhibitory signaling axes—such as STAT3/6 and PTEN/FOXO/AMPK—and in turn blocks fibrosis-related pathways, including MAPK, NF-κB, and mTOR pathways [ 10 , 15 , 16 , 17 , 18 ]. Existing studies have confirmed the definitive anti-inflammatory and antifibrotic potential of IL-37 across multiple organ systems. In pulmonary fibrosis models, IL-37 significantly alleviates bleomycin-induced pulmonary inflammatory infiltration and collagen deposition [ 19 ]. In models of diabetic nephropathy, IL-37 mitigates fatty acid oxidation dysfunction and renal fibrosis by upregulating CPT1A [ 20 ]. Furthermore, in diabetic cardiomyopathy, IL-37 improves mitochondrial damage and inhibits myocardial fibrosis by maintaining the function of the SIRT1–AMPK–PGC1α axis [ 21 ]. IL-37 not only alleviates inflammation and reverses fibrosis but also regulates macrophage polarization. Studies have shown that in diseases such as hepatitis, liver fibrosis, and periodontitis, IL-37 inhibits M1 polarization and promotes M2 polarization, thereby helping to modulate the immune balance [ 16 , 22 , 23 , 24 ]. In addition, in atherosclerosis, IL-37 suppresses macrophage ferroptosis through the NRF2 pathway [ 25 ]. Finally, in tumor immunity, it enhances macrophage phagocytic capacity via the IL-18Rα/SIGIRR-STAT3 axis [ 26 ]. Collectively, these effects provide a critical foundation for the role of IL-37 in mitigating tissue damage. The evidence presented indicates that IL-37 plays a pivotal role in regulating immunity and inhibiting fibrosis; however, its specific functions and therapeutic potential in CE remain unclear. In this study, we demonstrated that IL-37 exerts a protective effect on endometrial repair by inhibiting EMT and reprogramming macrophages toward M2 polarization, potentially coactivating the STAT6 and Smad3 signaling pathways. These findings may provide novel experimental evidence and strategies for the treatment of CE.

To evaluate the therapeutic potential of IL-37 for CE, we successfully expressed and purified the recombinant IL-37 protein, which was injected intraperitoneally into the rats to treat CE. Immunofluorescence analysis of uterine tissues revealed that, compared with the control group, the LPS-induced model group showed no significant change in IL-37 expression. By contrast, the IL-37 (1 μg and 2 μg) groups exhibited a marked increase in IL-37-specific fluorescence signals, which were primarily localized at the cell periphery ( Figure 1 A,B). These findings indicate that the exogenous IL-37 protein can be effectively taken up by rat uterine cells. Gross morphological observation revealed that, compared with the control group, the LPS group exhibited significant pathological changes in the uterus, including intrauterine fluid accumulation, tissue edema, and thinning of the uterine wall with increased transparency. However, following intervention with IL-37 (1 μg and 2 μg), these pathological manifestations were markedly alleviated, as evidenced by reduced intrauterine fluid and the restoration of normal uterine morphology ( Figure 1 A). H&E staining results ( Figure 1 A,C,D) revealed that the LPS group exhibited a damaged endometrial structure, along with extensive inflammatory cell infiltration and stromal edema. IL-37 treatment significantly attenuated inflammatory infiltration and edema, promoting the repair of the endometrial structure, which was reflected by the restoration of the endometrial area ratio and glandular number. Furthermore, Masson staining ( Figure 1 A,E) revealed that the LPS group exhibited significantly increased collagen fiber deposition, whereas the IL-37 intervention effectively reduced the area of collagen deposition and reversed the fibrotic process. Therefore, we concluded that exogenous IL-37 can be effectively delivered to injured uteri and can have therapeutic effects, including anti-inflammatory effects, tissue repair promotion, and antifibrotic activity. On the basis of previous findings that IL-37 can reverse endometrial fibrosis, we hypothesized that its effects may be associated with the inhibition of the EMT process. To evaluate the occurrence of EMT and the effects of IL-37 treatment at the tissue level, we conducted immunofluorescence staining of rat uterine tissues. The results ( Figure 2 ) indicated that, compared with the control group, the LPS group exhibited significantly downregulated expression of the epithelial marker E-cadherin and significantly upregulated expression of the mesenchymal marker vimentin in uterine tissues. Following IL-37 treatment, the fluorescence intensity of E-cadherin was effectively restored, whereas the expression of vimentin was significantly inhibited. Moreover, we conducted in vitro validation using LPS-stimulated rat renal tubular epithelial cells. Transwell migration assays demonstrated that IL-37 treatment effectively inhibited the LPS-induced increase in cell migration capacity ( Figure 3 A,B). Western blot and RT-qPCR results confirmed that IL-37 treatment reversed the dysregulated expression of key EMT markers induced by LPS. Specifically, IL-37 restored the expression of the epithelial marker E-cadherin and decreased the expression of several mesenchymal markers, including vimentin and N-cadherin ( Figure 3 C,D). These findings provide a critical mechanistic basis for the role of IL-37 in ameliorating endometrial fibrosis. To elucidate the regulatory role of IL-37 in the immune microenvironment of CE, we systematically evaluated its effects on macrophage polarization. First, immunofluorescence staining of uterine tissue revealed that LPS induced a proinflammatory environment characterized by the expansion of CD86 + M1 macrophages and the contraction of CD206 + M2 macrophages. The administered IL-37 dose-dependently restored this ratio, as it suppressed CD86 + fluorescence while amplifying CD206 + signals, with maximal restoration achieved at 2 µg of IL-37 ( Figure 4 ). Next, we assessed the effect of IL-37 on macrophage polarization in RAW264.7 cells. In LPS-driven M1 cultures, IL-37 decreased the transcription of IL-6, iNOS, and IL-1β and inhibited the expression of the master M1 transcription factor STAT3 ( Figure 5 A). Conversely, under IL-4-driven M2 conditions, IL-37 combined with IL-4 to increase the expression of TGF-β, CD206, and Arg1 while increasing the expression of STAT6 mRNA ( Figure 5 B). Flow cytometric quantification confirmed that the combination of IL-37 and IL-4 synergistically increased the CD206 + subpopulation in a concentration-dependent manner ( Figure 5 C). Collectively, these data establish that IL-37 suppresses the proinflammatory M1 phenotype while promoting the reparative M2 phenotype, thereby reprogramming the endometrial immune niche toward a reparative phenotype in CE. To determine whether IL-37 reprograms macrophage function by modulating the STAT6 and Smad3 signaling pathways, we conducted a systematic investigation of its effects on an IL-4-induced M2 polarization model. Western blot analysis revealed that treatment with IL-4 alone elevated the levels of phosphorylated STAT6 (p-STAT6) and IL-37 at concentrations ranging from 10 to 1000 ng/mL, dose-dependently increased the level of p-STAT6 and independently triggered robust phosphorylation of Smad3 (p-Smad3). Consequently, the ratios of p-STAT6 to total STAT6 and p-Smad3 to total Smad3 significantly increased, indicating synergistic potentiation of STAT6 and de novo activation of Smad3 ( Figure 6 A,B). Moreover, the results obtained with a Smad-responsive luciferase reporter (pLenti-Smad-minP-Luc) demonstrated that IL-37 increased luciferase activity in a concentration-dependent manner, indicating the amplification of Smad-mediated transcription ( Figure 6 C). To further visualize the changes in cell dynamics during this process, we performed immunofluorescence staining analysis and quantitatively evaluated the nuclear-translocation of STAT6 and Smad3, as shown in Figure 6 D. At 0 min of IL-37 treatment, STAT6 and Smad3 signals were predominantly localized in the cytoplasm. After 30 min of treatment, the nuclear-to-cytoplasmic ratio of both proteins was significantly increased compared to 0 min. At 1 h of treatment, the nuclear-to-cytoplasmic ratio further increased, showing a significant difference compared to 30 min. This time-dependent nuclear accumulation confirms IL-37’s effective induction of nuclear translocation for these two key transcription factors. In summary, IL-37 functions as a dual coactivator: it amplifies IL-4-triggered STAT6 signaling while independently activating the Smad3 cascade, thereby enforcing alternative M2 macrophage activation.

The endometrium is a cyclically reprogrammed mucosal surface whose capacity for scarless restoration underpins fertility. When this process is disrupted, CE and intrauterine adhesion (IUA) occur, resulting in the transformation of a transient inflammatory insult into a self-amplifying fibrotic circuit [ 31 , 32 , 33 ]. This study systematically elucidates, for the first time, the dual protective role of IL-37 in promoting endometrial injury repair: it directly inhibits EMT to preserve epithelial integrity and reprograms macrophage polarization to enhance the repair microenvironment through coordinated activation of the STAT6/Smad3 axis. These two mechanistic pathways collectively provide robust experimental evidence supporting the role of IL-37 as a potential novel therapeutic target for CE. IL-37, a member of the interleukin-1 family, exhibits extensive anti-inflammatory and immunoregulatory properties [ 34 ]. These characteristics confer protective effects in various disease models, including cardiovascular diseases, autoimmune disorders, and tumors [ 35 , 36 , 37 , 38 ]. In recent years, the role of IL-37 in diseases of the female reproductive system has garnered increasing attention. For instance, in endometriosis (EMS), recombinant human IL-37 can inhibit the growth of ectopic lesions by suppressing STAT3 phosphorylation in dendritic cells and modulating the Th1/Th2 balance [ 39 ]. In endometrial cancer, IL-37b targets the Rac1/NF-κB signaling pathway to reduce the expression of matrix metalloproteinase 2 (MMP2), effectively suppressing cancer cell migration and invasion [ 40 ]. Furthermore, endometrial regenerative cells that secrete IL-37 can significantly downregulate the expression of proinflammatory factors such as TNF-α, IL-1β, and IL-6 and upregulate the expression of IL-10, thereby exerting therapeutic effects in chronic transplant vasculopathy [ 41 ]. Our study is the first to demonstrate that exogenous IL-37 alleviates endometritis by inhibiting EMT and promoting M2 polarization. These findings collectively reveal the crucial regulatory role of IL-37 in both inflammatory and tumor-related pathological processes within the female reproductive system. EMT is a pivotal event in the fibrotic process of various organs and is characterized by the downregulation of the expression of epithelial markers, such as E-cadherin, and the upregulation of the expression of stromal markers, including N-cadherin and vimentin [ 42 ]. Maintaining epithelial cell homeostasis is crucial for preventing fibrosis [ 43 ], and inhibiting EMT serves as a common mechanism underlying the protective effects of IL-37 in several fibrotic diseases. In hepatic fibrosis, the intracellular form of IL-37 directly interacts with Smad3, thereby suppressing TGF-β signaling and hepatic stellate cell activation [ 16 ]. In asthma models, IL-37 alleviates airway EMT by modulating the ERK1/2 and STAT3 pathways [ 44 ]. Furthermore, in diabetic cardiomyopathy, IL-37 exerts antifibrotic effects by inhibiting the JAK2-STAT3 axis [ 45 ]. In CE, microbial-associated molecular patterns, such as LPS, drive the release of proinflammatory factors, including TNF-α and IL-6, as well as profibrotic factors, such as TGF-β, through the activation of the TLR4/NF-κB signaling axis [ 46 , 47 , 48 ]. IL-37 has been shown to effectively suppresses the activation of the NF-κB pathway [ 34 ], which may inhibit this upstream inflammatory signaling and downregulate the regulatory network that drives EMT. Our study confirms that IL-37 can reverse EMT in CE. In vivo, IL-37 restored the epithelial phenotype of the damaged endometrium. In vitro, it inhibited the LPS-induced migration of renal tubular epithelial cells while simultaneously reversing the abnormal expression of EMT markers. These findings indicate that IL-37 can prevent the initiation of fibrosis at its source by directly stabilizing epithelial cells, laying the foundation for tissue regeneration. As pivotal regulatory cells within the immune microenvironment of CE, macrophages directly influence the processes of endometrial tissue damage and repair through their polarization states [ 49 , 50 , 51 , 52 ]. Studies have demonstrated that modulating macrophage polarization can serve as an effective therapeutic strategy for CE. For instance, glycyrrhizic acid can alleviate endometrial hyperplasia and inflammation in CE mice by inhibiting HMGB1-mediated macrophage pyroptosis [ 51 ]. Adipose-derived stem cell exosomes suppress M1 polarization by regulating the SIRT2/NLRP3 axis, thereby mitigating endometritis [ 53 ]. Additionally, recombinant humanized type III collagen (rhCol III) has been shown both in vitro and in rat CE models to promote macrophage polarization toward the M2 phenotype by inhibiting the NF-κB/YAP pathway, consequently improving inflammation and facilitating the restoration of the immune microenvironment [ 6 ]. Although IL-37 has been established as a crucial immunoregulatory factor in various inflammatory models, its specific molecular mechanism in regulating macrophage polarization in chronic inflammation remains unclear. This study is the first to demonstrate that IL-37 can induce ‘functional reprogramming’ of macrophages in chronic inflammation models. The results indicate that under M1 polarization conditions, IL-37 significantly suppresses the expression of proinflammatory genes such as IL-6, iNOS, and IL-1β. Conversely, under M2 polarization conditions, IL-37 not only further upregulates the expression of classic M2 markers, including TGF-β, Arg1, and CD206, but also increases the proportion of CD206+ M2 cells. In-depth mechanistic studies revealed that IL-37 treatment significantly increased the phosphorylation levels of STAT6 and Smad3, thereby activating the downstream Smad signaling pathway to promote the expression of the repair-related protein ARG1. Notably, classical M2 macrophages typically activate the Smad pathway through TGF-β secretion, promoting EMT and fibrosis [ 54 , 55 ]. However, this study found that IL-37 significantly upregulated p-Smad3 levels and promoted its nuclear translocation, while simultaneously inhibiting EMT and fibrosis. This may initially seem contradictory; however, studies on tumor cells indicate that, unlike the TGF-β/Smad3 signaling pathway, IL-37 can directly bind to Smad3 intracellularly. This interaction inhibits the formation and nuclear translocation of the Smad2/3/4 complex, thereby attenuating the transcriptional activity of the canonical Smad pathway [ 56 ]. Additionally, the functional output of Smad3 is phospho-site-dependent. C-terminal phosphorylation (pSmad3C) primarily mediates growth inhibition and differentiation regulation, while linker region phosphorylation (pSmad3L) strongly drives extracellular matrix deposition and fibrotic transcriptional programs [ 16 ]. IL-37 achieves “functional remodeling” of the Smad3 signalling by specifically inhibiting the pSmad3L/c-Myc signalling axis while preserving or redirecting the pSmad3C pathway. This mechanism has been preliminarily validated in liver fibrosis models [ 24 ]. Therefore, we propose that IL-37 does not simply activate Smad3 but may instead reshape its phosphor–isomer balance and transcriptional output, enabling it to synergise with STAT6 to guide anti-inflammatory repair programs represented by ARG1, rather than the classical fibrotic gene programs. However, the exact regulatory mechanisms require further investigation. This study presents several limitations. First, while the NRK-52e renal tubular epithelial cell line serves as a classical model for elucidating EMT mechanisms, it does not fully replicate the distinct tissue microenvironment associated with endometritis. Future research should aim to validate these findings using human endometrial epithelial cell lines or primary cells to enhance the disease-specific relevance of this mechanism. Additionally, the in vitro macrophage polarisation experiment employed the mouse RAW 264.7 cell line, which differs from the rat model used in in vivo studies. Although the core signalling pathways are highly conserved across species, and this cell line offers the advantage of a mature experimental system, further validation using primary rat or bone marrow-derived macrophages is required to more accurately reflect the immunomodulatory mechanisms of IL-37 in CE models. Furthermore, the Smad reporter gene experiment utilized HEK-293T cells, primarily due to their high transfection efficiency and robust stability of the reporter system. Therefore, these findings serve as preliminary mechanistic evidence which requires further validation in disease-associated target cells for future studies. In summary, this study proposes an integrated mechanistic model of the role of IL-37 in promoting endometrial repair. In the CE-damaged microenvironment, IL-37 operates through dual synergistic pathways. On the one hand, it directly inhibits EMT to stabilize epithelial architecture; on the other hand, it reprograms macrophages by coordinately activating and remodeling the functional output of the STAT6/Smad3 axis, thereby shaping an efficient and safe pro-repair immune microenvironment. These two pathways mutually reinforce each other to collectively drive high-quality tissue regeneration. Although the precise mechanism through which IL-37 coordinately activates STAT6 and Smad3 remains to be elucidated further, this study provides a solid theoretical foundation and experimental basis for the development of novel therapies targeting IL-37 or its downstream signaling nodes for the treatment of CE and related endometrial fibrotic diseases.

Our results demonstrated that IL-37 accumulated in injured uterine tissue and significantly ameliorated inflammatory infiltration, tissue destruction, and collagen deposition. Treatment with IL-37 attenuated epithelial cell migration and reversed abnormal EMT by upregulating E-cadherin expression and downregulating vimentin expression. Furthermore, IL-37 inhibited M1 macrophage infiltration and promoted a shift toward the M2 phenotype. Mechanistic studies revealed that IL-37 coactivated the STAT6 and Smad3 signaling pathways, increasing their phosphorylation and nuclear translocation, which subsequently boosted ARG1 expression. These findings indicate that IL-37 ameliorates CE by attenuating EMT and promoting M2 macrophage polarization through coordinated activation of the STAT6/Smad3 axis, underscoring its potential as a therapeutic agent for CE.