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Suppression of Pro-fibrotic Signaling Potentiates Factor-mediated Reprogramming of Mouse Embryonic Fibroblasts into Induced Cardiomyocytes
Published on: June 3, 2018
miR-140-5p attenuates hepatic fibrosis by directly targeting TGFβR1
Wenchao Pan1, Yadong Wang1, Caiyan Zhao1
1Department of Infectious Diseases, The Third Hospital of Hebei Medical University, Shijiazhuang, China.
Objective:
To explore the protective effect and related mechanism of miR-140-5p on liver fibrosis by interfering with TGF-β/Smad signaling pathway.
Methods:
Liver fibrosis mice models were established by intraperitoneal injection of CCL4. Hematoxylin and eosin (HE) staining was used to detect the structural and morphological changes of the liver. Masson staining was used to detect collagen deposition. Human hepatic stellate cells (HSCs, LX-2) were transfected with miR-140-5p mimic or inhibitor then treated with TGF-β1. The qRT-PCR and Western blotting was used to detect the expression of related molecules. The luciferase reporter assay was used to identify the target of miR-140-5p.
Results:
Our results indicated that miR-140-5p expression was downregulated in fibrotic liver tissues of model mice and LX-2 cells treated with TGF-β1. The overexpression of miR-140-5p decreased the expression of collagen1(COL1) and α-smooth muscle actin(α-SMA), inhibited the phosphorylation of Smad-2/3 (pSmad-2/3) in LX-2 cells. Conversely, the knockdown of miR-140-5p upregulated COL1 and α-SMA expression, increased Smad-2/3 phosphorylation. A dual-luciferase reporter assay showed that TGFβR1 was a target gene of miR-140-5p. The overexpression of miR-140-5p suppressed TGFβR1 expression in LX-2 cells. Additionally, knockdown of TGFβR1 decreased the expression of COL1 and α-SMA. Conversely, the overexpression of TGFβR1 reversed the inhibitory effect of miR-140-5p upregulation on expression of COL1 and α-SMA.
Conclusion:
miR-140-5p bound to TGFβR1 mRNA 3'-untranslated region(3'UTR) and inhibited the expression of TGFβR1, pSmad-2/3, COL1 and α-SMA, thereby exerting a potential therapeutic effect on hepatic fibrosis.
Insights
MicroRNA-140-5p (miR-140-5p) protects against liver fibrosis by targeting TGFβR1 and inhibiting the TGF-β/Smad pathway. This finding offers a potential therapeutic strategy for liver fibrosis.
Area of Science:
- Hepatology
- Molecular Biology
- Biochemistry
Background:
- Liver fibrosis is a significant health concern characterized by excessive extracellular matrix deposition.
- The transforming growth factor-beta (TGF-β)/Smad signaling pathway plays a crucial role in the pathogenesis of liver fibrosis.
- MicroRNAs (miRNAs) are emerging as key regulators in liver fibrosis, but their specific roles and mechanisms require further elucidation.
Purpose of the Study:
- To investigate the protective role of miR-140-5p in liver fibrosis.
- To elucidate the underlying mechanism involving the TGF-β/Smad signaling pathway.
Main Methods:
- Established carbon tetrachloride (CCl4)-induced liver fibrosis mouse models.
- Utilized human hepatic stellate cells (LX-2) for in vitro experiments.
- Performed qRT-PCR, Western blotting, Masson staining, and dual-luciferase reporter assays.
Main Results:
- miR-140-5p was downregulated in fibrotic liver tissues and TGF-β1-treated LX-2 cells.
- Overexpression of miR-140-5p reduced collagen deposition and inhibited TGF-β/Smad signaling.
- TGFβR1 was identified as a direct target of miR-140-5p, mediating its antifibrotic effects.
Conclusions:
- miR-140-5p exerts a protective effect against liver fibrosis.
- This effect is mediated by the inhibition of TGFβR1 expression and subsequent suppression of the TGF-β/Smad signaling pathway.
- miR-140-5p represents a potential therapeutic target for liver fibrosis.
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