Department of Hepatobiliary and Pancreatic Surgery, Shenzhen People's Hospital (The Second Clinical Medical College, Jinan University, Guangzhou, China.
The First Affiliated Hospital, Southern University of Science and Technology), Shenzhen, China.
J Cell Mol Med. 2021 Jan;25(2):701-715. doi: 10.1111/jcmm.16119. Epub 2020 Dec 20.
Hepatic fibrosis (HF) is involved in aggravated wound-healing response as chronic liver injury. Extracellular vesicles (EVs) carrying microRNA (miR) have been reported as therapeutic targets for liver diseases. In this study, we set out to explore whether adipose-derived mesenchymal stem cells (ADMSCs)-derived EVs containing miR-150-5p affect the progression of HF. Carbon tetrachloride (CCl ) was firstly used to induce HF mouse models in C57BL/6J mice, and activation of hepatic stellate cells (HSCs) was achieved using transforming growth factor β (TGF-β). EVs were then isolated from ADMSCs and co-cultured with HSCs. The relationship between miR-150-5p and CXCL1 was identified using dual luciferase gene reporter assay. Following loss- and gain-function experimentation, HSC proliferation was examined by MTT assay, and levels of fibrosis-, HSC activation- and apoptosis-related genes were determined in vitro. Additionally, pathological scores, collagen volume fraction (CVF) as well as levels of inflammation- and hepatic injury-associated genes were determined in in vivo. Down-regulated miR-150-5p and elevated CXCL1 expression levels were detected in HF tissues. ADMSCs-derived EVs transferred miR-150-5p to HSCs. CXCL1 was further verified as the downstream target gene of miR-150-5p. Moreover, ADMSCs-EVs containing miR-150-5p markedly inhibited HSC proliferation and activation in vitro. Meanwhile, in vivo experiments also concurred with the aforementioned results as demonstrated by inhibited CVF, reduced inflammatory factor levels and hepatic injury-associated indicators. Both experiments results were could be reversed by CXCL1 over-expression. Collectively, our findings indicate that ADMSCs-derived EVs containing miR-150-5p attenuate HF by inhibiting the CXCL1 expression.
肝纤维化（HF）是慢性肝损伤导致的加重的伤口愈合反应。已经有研究报道，携带 microRNA（miR）的细胞外囊泡（EVs）是肝脏疾病的治疗靶点。在本研究中，我们旨在探讨脂肪间充质干细胞（ADMSCs）衍生的含有 miR-150-5p 的 EV 是否影响 HF 的进展。首先，我们用四氯化碳（CCl ）诱导 C57BL/6J 小鼠的 HF 模型，并用转化生长因子 β（TGF-β）激活肝星状细胞（HSCs）。然后从 ADMSCs 中分离 EV，并与 HSCs 共培养。通过双荧光素酶基因报告基因检测鉴定 miR-150-5p 与 CXCL1 之间的关系。通过失活和获得功能实验，通过 MTT assay 检测 HSC 增殖，在体外测定纤维化、HSC 激活和凋亡相关基因的水平。此外，在体内测定病理评分、胶原体积分数（CVF）以及炎症和肝损伤相关基因的水平。HF 组织中检测到 miR-150-5p 下调和 CXCL1 表达水平升高。ADMSCs 衍生的 EV 将 miR-150-5p 转染至 HSCs。进一步验证 CXCL1 是 miR-150-5p 的下游靶基因。此外，ADMSCs-EVs 中含有 miR-150-5p 可显著抑制 HSC 的增殖和体外激活。同时，体内实验结果也与上述结果一致，表现为 CVF 降低、炎症因子水平降低和肝损伤相关指标降低。这些结果都可以通过 CXCL1 过表达逆转。总之，我们的研究结果表明，ADMSCs 衍生的含有 miR-150-5p 的 EV 通过抑制 CXCL1 的表达来减轻 HF。
