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目的 探讨髓系细胞触发受体2(TREM2)介导的巨噬细胞极化对宫腔粘连(IUA)小鼠子宫内膜纤维化的影响。方法 选取8周龄SPF级雌性C57BL/6小鼠,Trem2基因敲除(Trem2 KO)和同窝野生型(WT)小鼠各5只,采用机械搔刮损伤法建立IUA模型,分别为Trem2 KO-IUA组和WT-IUA组,另选取5只WT小鼠作为Control组,术后约16d进行子宫组织取材分析。HE染色观察小鼠子宫内膜病理变化,Masson染色观察子宫内膜纤维化情况,流式细胞术检测内膜组织中TREM2+巨噬细胞数量,荧光定量-聚合酶链式反应(RT-PCR)检测基因表达情况,酶联免疫吸附试验(ELISA)检测巨噬细胞中炎症因子分泌水平。分离小鼠子宫内膜组织中巨噬细胞与人子宫内膜间质细胞(HESCs)进行共培养,EdU检测细胞增殖、流式细胞术检测细胞凋亡并通过细胞划痕实验检测迁移能力。结果 与Control组比较,WT-IUA组小鼠子宫宫腔出现明显粘连,内膜组织中TREM2+巨噬细胞数量显著增加,Trem2 mRNA相对表达量显著升高(P<0.001)。与WT-IUA组比较,Trem2 KO-IUA组小鼠内膜组织中与胶原沉积相关的Col1a1、α-Sma mRNA相对表达量均显著增加(P<0.05),与炎症相关的Ccl2 mRNA相对表达量也显著增加(P<0.01)。从WT-IUA组和Trem2 KO-IUA组小鼠内膜组织中分离出的巨噬细胞与HESCs共培养后,与WT-IUA组比较,Trem2 KO-IUA组巨噬细胞分泌IL-1β和TNF-α蛋白水平均显著增加(P<0.01),而IL-10水平显著降低(P<0.01)。RT-PCR结果显示,与WT-IUA组相比,Trem2 KO-IUA组巨噬细胞极化相关基因Cd86 mRNA相对表达量显著增加(P<0.01),而Cd206 mRNA相对表达量显著降低(P<0.001)。与WT-IUA组比较,Trem2 KO-IUA组共培养的HESCs增殖能力显著增加(P<0.001),细胞凋亡数量显著减少(P<0.001),细胞迁移能力显著增强(P<0.01),HESCs中α-Sma和Col1a1 mRNA相对表达量均显著增加(P<0.01)。结论 TREM2可能通过促进IUA小鼠子宫内膜巨噬细胞M2型极化,抑制炎症反应,抑制HESCs异常活化,减少宫腔胶原沉积,从而缓解IUA小鼠子宫内膜纤维化。TREM2可成为IUA的潜在治疗靶点。
Abstract：Objectives:To investigate the effect of triggering receptor expressed on myeloid cells 2(TREM2)-mediated macrophage polarization on endometrial fibrosis in mice with intrauterine adhesion(IUA).Methods:Eight-week-old specific pathogen-free(SPF)female C57BL/6 mice,including Trem2 knockout(Trem2 KO)mice and their wild-type(WT)littermates(n=5 per group),were selected.An IUA model was established using a standard mechanical scraping injury method,resulting in the Trem2 KO-IUA group and the WT-IUA group.An additional five WT mice were used as the control(sham-operated)group.Uterine tissues were collected approximately 16 days post-surgery for analysis.Hematoxylin and eosin(HE)staining was used to observe endometrial pathological changes,and Masson's staining was employed to assess endometrial fibrosis.The number of TREM2~+macrophages in the endometrial tissue was quantified by flow cytometry.Reverse transcription-polymerase chain reaction(RT-PCR)was performed to measure the mRNA expression levels.Enzyme-linked immunosorbent assay(ELISA)was used to detect the secretion levels of inflammatory factors.Macrophages isolated from the endometrial tissues were co-cultured with human endometrial stromal cells(HESCs).Subsequently,an EdU assay was conducted to evaluate cell proliferation,flow cytometry was used to assess apoptosis,and a wound healing assay was performed to measure migration capacity.Results:Compared with the control group,the WT-IUA group exhibited significant uterine cavity adhesions,a marked increase in the number of TREM2~+macrophages,and a significantly higher relative expression of Trem2 mRNA in the endometrial tissue(P<0.001).Compared with the WT-IUA group,the Trem2 KO-IUA group showed significantly increased relative expression of fibrosis-related genes Col1a1 and α-Sma mRNA(P <0.05),as well as a significantly increased relative expression of the inflammation-related gene Ccl2 mRNA(P<0.01).Macrophages isolated from the endometrial tissues of WT-IUA and Trem2 KO-IUA groups were co-cultured with HESCs and divided into the WT-IUA group and the Trem2 KO-IUA group.The secretion levels of interleukin(IL)-1β and tumor necrosis factor(TNF)-α protein by macrophages in the Trem2 KO-IUA group was significantly higher than that in the WT-IUA group(P<0.01),while IL-10 level was significantly lower than that in the WT-IUA group(P<0.01).RT-PCR results indicated that compared with the WT-IUA group,macrophages in the Trem2 KO-IUA group had a significantly higher relative expression of the M1 marker Cd86 mRNA(P<0.01)and a significantly lower relative expression of the M2 marker Cd206 mRNA(P<0.001).Furthermore,HESCs co-cultured with Trem2 KO-IUA macrophages demonstrated significantly enhanced proliferation(P<0.001),significantly reduced apoptosis(P<0.001),and significantly increased migration capacity(P<0.01)compared to the WT-IUA group.The relative expression of α-Sma and Col1a1 mRNA in these HESCs were also significantly increased(P<0.01).Conclusions:TREM2 may alleviate endometrial fibrosis in IUA mice by promoting M2 polarization of endometrial macrophages,inhibiting inflammatory responses,suppressing the abnormal activation of HESCs,and reducing collagen deposition in the uterine cavity.Therefore,TREM2 represents a potential therapeutic target for IUA.
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基本信息:
中图分类号:R711.74
引用信息:
[1]沈聪,沙晨阳,杨梦琪,等.TREM2通过调控巨噬细胞M2极化抑制宫腔粘连小鼠子宫内膜纤维化的作用及机制研究[J].生殖医学杂志,2025,34(12):1665-1673.
基金信息:
国家自然科学基金青年项目（82305283）; 四明基金（SGKY-202407）; 上海市浦东新区卫生健康委员会学科建设计划项目（PWZzb2022-25）
2025-05-04
2025
2025-07-22
2025-11-14
2025
1
2025-12-15
2025-12-15