Purpose

Telocytes (TCs), a novel type of stromal cells found in tissues, induce macrophage differentiation into classically activated macrophages (M1) types and enhance their phagocytic function. The purpose of this study was to investigate the inhibitory effects of TC-induced M1 macrophages on endometriosis (EMs).

Methods

mouse uterine primary TCs and endometrial stromal cells (ESCs) were isolated and identified using double immunofluorescence staining. For the in vitro study, ESCs were treated with TC-induced M1 macrophages, and the vascular endothelial growth factor (VEGF), matrix metalloproteinase 9 (MMP9), and nuclear factor kappa B (NF-κb) genes were identified by quantitative real-time PCR (qRT-PCR) or western blotting (WB). For the in vivo study, an EMs mouse model received TC-conditioned medium (TCM) via abdominal administration, and characterized the inhibitory effects on growth (lesion weight, volume, and pathology), tissue-resident macrophages differentiation by immunostaining, angiogenic capacity (CD31 and VEGF), invasive capacity (MMP9), and NF-κb expression within EMs lesions.

Results

immunofluorescent staining showed that uterine TCs expressed CD34+ and vimentin+, whereas ESCs expressed vimentin+ and cytokeratin-. At the cellular level, TC-induced M1 macrophages can significantly inhibit the expression of VEGF and MMP9 in ESCs through WB or qRT-PCR, possibly by suppressing the NF-κb pathway. The in vivo study showed that macrophages switch from the alternatively activated macrophages (M2) in untreated EMs lesions to the M1 subtype after TCM exposure. Thereby, TC-induced M1 macrophages contributed to the inhibition of EMs lesions. More importantly, this effect may be achieved by suppressing the expression of NF-κb to inhibit angiogenesis (CD31 and VEGF) and invasion (MMP9) in the tissue.

Conclusion

TC-induced M1 macrophages play a prevailing role in suppressing EMs by inhibiting angiogenic and invasive capacity through the NF-κb pathway, which provides a promising therapeutic approach for EMs.

中文翻译：

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端细胞诱导的 M1 巨噬细胞对子宫内膜异位症的抑制作用：靶向血管生成和侵袭

目的

特洛细胞（TC）是组织中发现的一种新型基质细胞，可诱导巨噬细胞分化为经典激活的巨噬细胞（M1）类型并增强其吞噬功能。本研究的目的是探讨TC诱导的M1巨噬细胞对子宫内膜异位症（EMs）的抑制作用。

方法

使用双重免疫荧光染色分离并鉴定小鼠子宫原代 TC 和子宫内膜基质细胞 (ESC)。在体外研究中，用 TC 诱导的 M1 巨噬细胞处理 ESC，并通过定量实时鉴定血管内皮生长因子 ( VEGF )、基质金属蛋白酶 9 ( MMP9 ) 和核因子 kappa B ( NF-κb ) 基因。时间 PCR (qRT-PCR) 或蛋白质印迹 (WB)。在体内研究中，EMs 小鼠模型通过腹部给药接受 TC 条件培养基 (TCM) ，并通过免疫染色表征对生长的抑制作用（病变重量、体积和病理学）、组织驻留巨噬细胞分化、血管生成能力（ CD31 和 VEGF)、侵袭能力 (MMP9) 和 EM 病变内的 NF-κb 表达。

结果

免疫荧光染色显示子宫TC表达CD34+和vimentin+，而ESC表达vimentin+和细胞角蛋白-。在细胞水平上，TC诱导的M1巨噬细胞可以通过WB或qRT-PCR显着抑制ESC中VEGF和MMP9的表达，可能是通过抑制NF-κb通路。体内研究表明，在接触中药后，未经治疗的 EM 病变中的巨噬细胞从替代激活的巨噬细胞 (M2) 转变为 M1 亚型。因此，TC 诱导的 M1 巨噬细胞有助于抑制 EM 病变。更重要的是，这种作用可能是通过抑制NF-κb的表达来抑制组织中的血管生成（CD31和VEGF）和侵袭（MMP9）来实现的。

结论

TC 诱导的 M1 巨噬细胞通过 NF-κb 途径抑制血管生成和侵袭能力，在抑制 EM 方面发挥着重要作用，这为 EM 提供了一种有前景的治疗方法。