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Extracellular Vesicles from miR-148a-5p-Enriched Bone Marrow Mesenchymal Stem Cells Relieve Hepatic Fibrosis by Targeting Smad4

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Abstract

Liver fibrosis is a hallmark feature of many chronic liver diseases, which is the leading cause of morbidity and mortality worldwide. Bone marrow mesenchymal stem cells (BMSCs)-derived extracellular vesicles have been applied in many diseases. In this study, we aimed to explore the specific mechanism of extracellular vesicles from BMSCs in liver fibrosis. Bioinformatics analysis was employed to screen miRNA and its target mRNA. Sirius Red staining was carried out to examine fibrosis in liver tissues. Extracellular vesicle morphology was assessed using Transmission Electron Microscopy. Quantitative real-time PCR (qRT-PCR) and western blotting analysis were performed to detect the expressions of miR-148a-5p, Smad4, transforming growth factor-β1 (TGF-β1), tissue inhibitor of metalloproteinase 1 (TIMP-1), Collagen I, α-smooth muscle actin (α-SMA), and extracellular vesicle markers CD9, TSG101, CD63, and calnexin. Dual-luciferase report gene assay was used for the luciferase activity analysis. Bioinformatics analysis revealed miR-148a-5p as a regulator in liver fibrosis. QRT-PCR results indicated that miR-148a-5p was lowly expressed in both thioacetamide (TAA)-induced mice and TGF-β1-activated hepatic stellate cells. Extracellular vesicles from miR-148a-5p enriched BMSCs downregulated the mRNA and protein levels of TGF-β1, TIMP-1, Collagen I, and α-SMA. Further bioinformatics analysis indicated that Smad4 was related to liver fibrosis. Furthermore, the dual-luciferase report gene assay confirmed the binding relationship between miR-148a-5p and Smad4. Extracellular vesicles from miR-148a-5p enriched BMSCs attenuated hepatic fibrosis in liver fibrosis by targeting Smad4.

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Funding

This work was supported by the ‘333 High-level Talents Project’ of Jiangsu Province (Grant No. BRA2018408), the Medical Scientific Research Project of Jiangsu Provincial Health Commission (Grant No. H2019073), and the Natural Science Foundation of Jiangsu Province (Youth Project, SBK2020041194).

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Author notes

  1. Ji Xuan, Huabin Xu, Hui Li and Desheng Chen have contributed equally to this work.

Authors and Affiliations

  1. Department of Gastroenterology, Jinling Hospital, Nanjing University, School of Medicine, Nanjing, 210002, Jiangsu, China

    Ji Xuan, Huabin Xu, Yuping Qiu, Xi Chen & Mei Shao

  2. Department of Gastroenterology, Affiliated Hospital of Integrated Traditional Chinese and Western Medicine, Nanjing University of Chinese Medicine, Nanjing, 210028, Jiangsu, China

    Hui Li

  3. Department of Blood Transfusion Medicine, Jinling Hospital, Nanjing University, School of Medicine, Nanjing, 210002, Jiangsu, China

    Desheng Chen

  4. Department of Digestive Medicine Center and General Practice, The Seventh Affiliated Hospital of Sun Yat-sen University, No. 628, Zhenyuan Rd., Xinhu Street, Guangming District, Shenzhen, 518107, Guangdong, China

    Xianming Xia

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  1. Ji Xuan
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12033_2021_441_MOESM1_ESM.jpg

Supplementary Figure 1 Culture and identification of primary BMSCs. (A) Morphological observation of primary BMSCs (scale bar: 100 μm). (B) The positive rates of CD105, CD73, and CD45 were analyzed by flow cytometry. (JPG 3551 kb)

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Xuan, J., Xu, H., Li, H. et al. Extracellular Vesicles from miR-148a-5p-Enriched Bone Marrow Mesenchymal Stem Cells Relieve Hepatic Fibrosis by Targeting Smad4. Mol Biotechnol 64, 535–545 (2022). https://doi.org/10.1007/s12033-021-00441-5

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