Objectives

Aim of this study is to explore whether quercetin can inhibit the enlarged fibrogenic responses of endometrial stromal cells by increasing the level of microRNA-145 (miR-145) and mediating the TGFβ1/Smad2/Smad3 signaling pathway, and to discuss the mechanism of signal transduction, further to provide experimental basis for revealing the pathophysiological mechanism and seeking new strategies for effective prevention and treatment of endometrial fibrosis.

Methods

The expression levels of miR-145 and TGF-β receptor 2 (TGFBR2) were detected by RT-qPCR analysis. Expressions of α-smooth muscle actin (α-SMA) and vimentin were examined by immunofluorescence staining. Cell viability was measured by MTT assay. The protein expression of collagen type 1 alpha 1 (Col1a1), α-SMA, fibronectin (FN), TGFBR2, transforming growth factor (TGF-β1), Smad2/3, phospho-Smad2/3 (p-Smad2/3) were detected by western blot analysis. The interaction between miR-145 and TGFBR2 was confirmed by dual-luciferase reporter gene assay.

Results

The expression level of miR-145 was decreased, whereas TGFBR2 was increased in intrauterine adhesion tissue. The expression levels of COL1A1, α-SMA, FN, TGFBR2, and p-Smad2/3 were increased, whereas miR-145 and cell proliferation were decreased in human endometrial stromal cells (hESCs) in response to TGF-β1 stimulation in a time and dose-dependent manner, which could be reversed by quercetin. Furthermore, quercetin regulates cell fibrogenic responses of endometrial stromal cells via miR-145/TGF-β1/Smad2/Smad3 pathway.

Conclusions

These findings indicated that quercetin have a significant anti-fibrotic effect and could upregulate miR-145 and inhibit activation of TGF-β1/Smad2/Smad3 pathway to regulate TGF-β1 induced fibrogenic responses of endometrial stromal cells, which may serve as a potential therapeutic agent for endometrial fibrosis.

中文翻译：

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槲皮素通过上调miR-145和抑制TGF-β1/Smad2/Smad3通路调节子宫内膜基质细胞的纤维化反应

目标

本研究的目的是探讨槲皮素是否可以通过增加微小RNA-145（miR-145）的水平和介导TGFβ1/Smad2/Smad3信号通路来抑制子宫内膜基质细胞扩大的纤维化反应，并讨论信号的机制进一步为揭示子宫内膜纤维化的病理生理机制和寻找有效防治子宫内膜纤维化的新策略提供实验依据。

方法

通过RT-qPCR分析检测miR-145和TGF-β受体2（TG​​FBR2）的表达水平。通过免疫荧光染色检查α-平滑肌肌动蛋白（α-SMA）和波形蛋白的表达。通过MTT测定测量细胞活力。1型胶原α1（Col1a1）、α-SMA、纤连蛋白（FN）、TGFBR2、转化生长因子（TGF-β1）、Smad2/3、磷酸-Smad2/3（p-Smad2/3）的蛋白表达分别为通过蛋白质印迹分析检测。双荧光素酶报告基因检测证实了 miR-145 和 TGFBR2 之间的相互作用。

结果

宫腔粘连组织中miR-145的表达水平降低，而TGFBR2的表达水平升高。COL1A1、α-SMA、FN、TGFBR2和p-Smad2/3的表达水平增加，而人子宫内膜基质细胞（hESCs）中miR-145和细胞增殖在一段时间内响应TGF-β1刺激而降低和剂量依赖性方式，这可以被槲皮素逆转。此外，槲皮素通过 miR-145/TGF-β1/Smad2/Smad3 通路调节子宫内膜基质细胞的细胞纤维化反应。

结论

这些发现表明槲皮素具有显着的抗纤维化作用，可以上调 miR-145 并抑制 TGF-β1/Smad2/Smad3 通路的激活，从而调节 TGF-β1 诱导的子宫内膜基质细胞的纤维化反应，可作为一种潜在的治疗方法。治疗子宫内膜纤维化的药物。