MicroRNA‑326 inhibits endometrial fibrosis by regulating TGF‑β1/Smad3 pathway in intrauterine adhesions.

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.