PMID- 35883205 OWN - NLM STAT- MEDLINE DCOM- 20220728 LR - 20220801 IS - 1757-6512 (Electronic) IS - 1757-6512 (Linking) VI - 13 IP - 1 DP - 2022 Jul 26 TI - Mesenchymal stem cells improve liver fibrosis and protect hepatocytes by promoting microRNA-148a-5p-mediated inhibition of Notch signaling pathway. PG - 354 LID - 10.1186/s13287-022-03030-8 [doi] LID - 354 AB - BACKGROUND: Mesenchymal stem cells (MSCs) are considered to be a potential therapeutic tool for liver fibrosis. Inhibiting the activation of hepatic stellate cells (HSCs) and protecting hepatocytes are important mechanisms for the anti-fibrotic effect of MSCs. However, how MSCs inhibit liver fibrosis by regulating the expression of microRNAs (miRNAs) has not been fully clarified. METHODS: Transforming growth factor-beta1 (TGF-beta1)-activated HSCs LX-2 were single cultured or co-cultured with human umbilical cord mesenchymal stem cells (HUC-MSCs). High-throughput sequencing was used to evaluate the differentially expressed microRNAs (DEMs) between the two groups. Quantitative real-time PCR (qRT-PCR), Western blot, and transfection experiments were used to investigate and screen the most significantly up-regulated DEM. Bioinformatics analysis was used to predict the target mRNAs and the potential functions of the DEM. The possible mechanism of HUC-MSCs against liver fibrosis was analyzed by co-culture experiment of HUC-MSCs with LX-2 cells, and HUC-MSCs treatment of Bile duct ligation (BDL)-induced liver fibrosis in mice. Finally, the mechanism of the DEM regulating liver fibrosis was confirmed in human liver fibrosis specimens. RESULTS: MicroRNA-148a-5p (miR-148a-5p) was the most significantly up-regulated DEM in activated LX-2 cells co-cultured with HUC-MSCs compared with LX-2 cells single cultured. Up-regulation of the expression of miR-148a-5p in activated LX-2 cells could significantly inhibit the expression of hepatic fibrosis markers alpha-SMA and Col1alpha1. Notch2 was one target gene of miR-148a-5p. Co-cultured with HUC-MSCs could inhibit the activation of LX-2 cells by inhibiting the expression of the Notch2 and the Notch signaling pathway. In addition, HUC-MSCs treatment could up-regulate the expression of miR-148a-5p in liver tissue and hepatocytes, promote the proliferation and avoid the apoptosis of hepatocytes, and reduce the degree of fibrosis by inhibiting expression of the Notch2 and the Notch signaling pathway in BDL-induced liver fibrosis mice. Moreover, miR-148a-5p was down-regulated and Notch2 was up-regulated in fibrotic human liver tissues compared with the normal livers. CONCLUSIONS: HUC-MSCs treatment could inhibit HSCs activation, protect hepatocytes, and alleviate BDL-induced liver fibrosis in mice by up-regulating the expression of miR-148-5p and inhibiting the Notch signaling pathway. The down-regulation of miR-148-5p and up-regulation of Notch2 could be used as biomarkers to monitor the progression of liver fibrosis. CI - (c) 2022. The Author(s). FAU - Zhou, Qing AU - Zhou Q AD - Department of Pathology, School of Biology and Basic Medical Sciences, Suzhou Medical College, Soochow University, Suzhou, 215123, China. AD - Department of General Surgery, Affiliated Hospital of Jiangsu University, Zhenjiang, 212001, China. FAU - Rong, Chao AU - Rong C AD - Department of Pathology, School of Biology and Basic Medical Sciences, Suzhou Medical College, Soochow University, Suzhou, 215123, China. FAU - Gu, Tengfei AU - Gu T AD - Department of Anesthesiology, People's Hospital of Lianshui County, Huaian, 223400, China. FAU - Li, Hongda AU - Li H AD - Department of General Surgery, Affiliated Hospital of Jiangsu University, Zhenjiang, 212001, China. FAU - Wu, Lei AU - Wu L AD - Department of Pathology, School of Biology and Basic Medical Sciences, Suzhou Medical College, Soochow University, Suzhou, 215123, China. FAU - Zhuansun, Xuemei AU - Zhuansun X AD - Department of Pathology, School of Biology and Basic Medical Sciences, Suzhou Medical College, Soochow University, Suzhou, 215123, China. FAU - Zhao, Xin AU - Zhao X AD - Department of Pathology, School of Biology and Basic Medical Sciences, Suzhou Medical College, Soochow University, Suzhou, 215123, China. FAU - Xiao, Zuorun AU - Xiao Z AD - Department of Pathology, School of Biology and Basic Medical Sciences, Suzhou Medical College, Soochow University, Suzhou, 215123, China. FAU - Kuang, Yuting AU - Kuang Y AD - Department of General Surgery, First Affiliated Hospital of Soochow University, Suzhou, 215008, China. FAU - Xu, Sanrong AU - Xu S AD - Department of General Surgery, Affiliated Hospital of Jiangsu University, Zhenjiang, 212001, China. zjxsrong@163.com. FAU - Wang, Shouli AU - Wang S AUID- ORCID: 0000-0002-6582-0726 AD - Department of Pathology, School of Biology and Basic Medical Sciences, Suzhou Medical College, Soochow University, Suzhou, 215123, China. wangsoly112@hotmail.com. LA - eng PT - Journal Article DEP - 20220726 PL - England TA - Stem Cell Res Ther JT - Stem cell research & therapy JID - 101527581 RN - 0 (MIRN148 microRNA, human) RN - 0 (MicroRNAs) RN - 0 (Mirn148 microRNA, mouse) SB - IM MH - Animals MH - Fibrosis MH - Hepatic Stellate Cells/metabolism MH - Hepatocytes/metabolism MH - Humans MH - Liver Cirrhosis/genetics/metabolism/therapy MH - *Mesenchymal Stem Cells/metabolism MH - Mice MH - *MicroRNAs/metabolism MH - Signal Transduction PMC - PMC9327397 OTO - NOTNLM OT - Hepatic stellate cell OT - Liver fibrosis OT - Mesenchymal stem cells OT - MicroRNA-148a-5p OT - Notch signaling pathway OT - Notch2 COIS- The authors declare that they have no conflicts of interest. EDAT- 2022/07/27 06:00 MHDA- 2022/07/29 06:00 PMCR- 2022/07/26 CRDT- 2022/07/26 23:53 PHST- 2022/05/17 00:00 [received] PHST- 2022/07/04 00:00 [accepted] PHST- 2022/07/26 23:53 [entrez] PHST- 2022/07/27 06:00 [pubmed] PHST- 2022/07/29 06:00 [medline] PHST- 2022/07/26 00:00 [pmc-release] AID - 10.1186/s13287-022-03030-8 [pii] AID - 3030 [pii] AID - 10.1186/s13287-022-03030-8 [doi] PST - epublish SO - Stem Cell Res Ther. 2022 Jul 26;13(1):354. doi: 10.1186/s13287-022-03030-8.