Third Clinical Medical College, Nanchang University, Nanchang, China; Department of Gastroenterology, The First Hospital of Nanchang (The Third Affiliated Hospital of Nanchang University), Nanchang, China.
Department of Gastroenterology, The First Hospital of Nanchang (The Third Affiliated Hospital of Nanchang University), Nanchang, China.
Cytokine. 2023 Dec;172:156386. doi: 10.1016/j.cyto.2023.156386. Epub 2023 Oct 16.
Human adipose-derived mesenchymal stem cell exosomes (ADSC-Exos) are active constituents for treating liver fibrosis. This paper attempted to preliminarily explain the functional mechanism of ADSC-Exos in liver fibrosis through the p38 MAPK/NF-κB pathway.
The cell models of hepatic fibrosis were established by inducing LX-2 cells with TGF-β1. Mouse models of liver fibrosis were established by treating mice with CCl. The in vivo and in vitro models of liver fibrosis were treated with ADSC-Exos. ADSCs were identified by flow cytometry/Alizarin red/oil red O/alcian blue staining. ADSC-Exos were identified by transmission electron microscopy, nanoparticle tracking analysis, and Western blot. LX-2 cell proliferation/viability were evaluated by MTT/BrdU assays. Exosomes were tracked in vivo and body weight changes in mice were monitored. Hepatic pathological changes were observed by HE/Masson staining. α-SMA/collagen I levels in liver tissues were assessed by immunohistochemistry. HA/PIIINP concentrations were measured using the magnetic particle chemiluminescence method. Liver function was assessed using an automatic analyzer. miR-20a-5p level was measured by RT-qPCR. The mRNA levels of fibrosis markers were determined by RT-qPCR, and their protein levels and levels of MAPK/NF-κB pathway-related proteins, as well as TGFBR2 protein level were measured by Western blot. The P65 nuclear expression in mouse liver tissues was quantified by immunofluorescence.
ADSC-Exos suppressed TGF-β1-induced LX-2 cell proliferation and fibrosis and reduced mRNA and protein levels of fibrosis markers in vitro. ADSC-Exos ameliorated liver fibrosis by inhibiting the p38 MAPK/NF-κB pathway activation. ADSC-Exos inhibited activation of the p38 MAPK/NF-κB pathway via regulating the miR-20a-5p/TGFBR2 axis. The in vivo experiment asserted that ADSC-Exos were mainly distributed in the liver, and ADSC-Exos relieved liver fibrosis in mice, which was evidenced by alleviating decreased body weight, reducing collagen and enhancing liver function, and repressed the activation of the p38 MAPK/NF-κB pathway via the miR-20a-5p/TGFBR2 axis.
ADSC-Exos attenuated liver fibrosis by suppressing the activation of the p38 MAPK/NF-κB pathway via the miR-20a-5p/TGFBR2 axis.
人脂肪间充质干细胞外泌体（ADSC-Exos）是治疗肝纤维化的有效成分。本文试图通过 p38MAPK/NF-κB 通路初步解释 ADSC-Exos 在肝纤维化中的功能机制。
用 TGF-β1 诱导 LX-2 细胞建立肝纤维化细胞模型。用 CCl 处理小鼠建立肝纤维化小鼠模型。用 ADSC-Exos 处理肝纤维化的体内和体外模型。通过流式细胞术/茜素红/oil red O/阿尔辛蓝染色鉴定 ADSC。通过透射电子显微镜、纳米颗粒跟踪分析和 Western blot 鉴定 ADSC-Exos。通过 MTT/BrdU 测定评估 LX-2 细胞增殖/活力。体内追踪外泌体，监测小鼠体重变化。通过 HE/Masson 染色观察肝组织的病理变化。通过免疫组化评估肝组织中α-SMA/胶原 I 水平。采用磁微粒化学发光法测定 HA/PIIINP 浓度。用自动分析仪评估肝功能。通过 RT-qPCR 测定 miR-20a-5p 水平。通过 RT-qPCR 测定纤维化标志物的 mRNA 水平，通过 Western blot 测定 MAPK/NF-κB 通路相关蛋白和 TGFBR2 蛋白水平，通过免疫荧光法定量测定小鼠肝组织中 P65 核表达。
ADSC-Exos 抑制 TGF-β1 诱导的 LX-2 细胞增殖和纤维化，并降低体外纤维化标志物的 mRNA 和蛋白水平。ADSC-Exos 通过抑制 p38MAPK/NF-κB 通路的激活来改善肝纤维化。ADSC-Exos 通过调节 miR-20a-5p/TGFBR2 轴抑制 p38MAPK/NF-κB 通路的激活。体内实验表明，ADSC-Exos 主要分布在肝脏，ADSC-Exos 减轻了小鼠的肝纤维化，表现为减轻体重下降、减少胶原和改善肝功能，并通过 miR-20a-5p/TGFBR2 轴抑制 p38MAPK/NF-κB 通路的激活。
ADSC-Exos 通过 miR-20a-5p/TGFBR2 轴抑制 p38MAPK/NF-κB 通路的激活来减轻肝纤维化。
