Zhou Hanyu, Wu Yanyan, Xue Junchao, Yu Liushenyan
Department of Pharmacy, Tongde Hospital of Zhejiang Province, Xihu District, Hangzhou City, Zhejiang Province, PR China.
Histol Histopathol. 2025 May;40(5):757-772. doi: 10.14670/HH-18-816. Epub 2024 Sep 18.
Hepatic fibrosis, ultimately causing hepatic sclerosis, remains significant health concerns. Adipose-derived mesenchymal stem cell (ADMSC)-derived exosomes (Exo) exhibit amelioration of liver injury. Hepatocyte growth factor (HGF) regulates hepatocyte growthn. However, its involvement during hepatic fibrosis remains unclear.
Isolation of ADMSCs and Exo, transfection of HGF overexpression, and activation of hepatic stellate cells (HSCs) by Angiotensin II (AngII) were conducted. Cells were randomized into HSC, AngII-HSC, ADMSCs-Exo, ADMSCs-Exo, and ADMSCs-Exo, DPI, LY294002, and SB203580 groups. MTT for cell viability, cell migration, and flow cytometry for ROS were performed. BALB/c mice were treated with CCL4 for hepatic fibrosis models. The mice were randomized into Control, PBS, ADMSCs-Exo, ADMSCs-Exo, and ADMSCs-Exo groups (n=6). HE, Sirius red, and Oil Red O staining, liver function indicators, and ELISA for oxidative stress were performed. ROS generation-related and PI3K/Akt/P38MAPK-related factors were detected by immunofluorescence, immunohistochemistry, and western blot.
After identification of ADMSC-Exo and transfection, AngII increased cell viability, migration, Collagen I (CoLI), α-smooth muscle actin (α-SMA), ROS, NADPH oxidase 4 (NOX4), PI3K, p-Akt, p-P38MAPK, ras-related C3 botulinum toxin substrate 1 (RAC1), p47, and p22 expression. However, ADMSCs-Exo, DPI, LY294002, and SB203580 reversed the above effects. Moreover, ADMSCs-Exo inhibited pathological damage, fibrosis, lipid accumulation, ALT, AST, TBIL, CoLI, α-SMA, NOX4, MDA, PI3K, P-Akt, and P-P38MAPK expression, and increased ALB, SOD, GPx, CAT, GSH, Mn-SOD, Na-K-ATPase, and Ca-Mg-ATPase levels in hepatic fibrosis mice.
ADMSCs-Exo attenuated hepatic fibrosis by inhibiting oxidative stress through activating the PI3K/Akt/P38MAPK pathway, providing valuable insights for potential treatment of liver fibrosis.
肝纤维化最终会导致肝硬化，仍然是严重的健康问题。脂肪来源的间充质干细胞（ADMSC）衍生的外泌体（Exo）可改善肝损伤。肝细胞生长因子（HGF）调节肝细胞生长。然而，其在肝纤维化过程中的作用仍不清楚。
进行ADMSC和Exo的分离、HGF过表达的转染以及用血管紧张素II（AngII）激活肝星状细胞（HSC）。将细胞随机分为HSC、AngII-HSC、ADMSCs-Exo、ADMSCs-Exo以及ADMSCs-Exo、DPI、LY294002和SB203580组。进行MTT法检测细胞活力、细胞迁移以及用流式细胞术检测活性氧（ROS）。用四氯化碳处理BALB/c小鼠建立肝纤维化模型。将小鼠随机分为对照组、PBS组、ADMSCs-Exo组、ADMSCs-Exo组和ADMSCs-Exo组（n = 6）。进行苏木精-伊红（HE）染色、天狼星红染色和油红O染色、肝功能指标检测以及氧化应激的酶联免疫吸附测定（ELISA）。通过免疫荧光、免疫组织化学和蛋白质印迹法检测与ROS生成相关以及与磷脂酰肌醇-3-激酶/蛋白激酶B/丝裂原活化蛋白激酶（PI3K/Akt/P38MAPK）相关的因子。
在鉴定ADMSC-Exo并进行转染后，AngII增加了细胞活力、迁移能力、I型胶原蛋白（CoLI）、α平滑肌肌动蛋白（α-SMA）、ROS、烟酰胺腺嘌呤二核苷酸磷酸氧化酶4（NOX4）、PI3K、磷酸化蛋白激酶B（p-Akt）、磷酸化P38丝裂原活化蛋白激酶（p-P38MAPK）、Rac家族小G蛋白1（RAC1）、p47和p22的表达。然而，ADMSCs-Exo、DPI、LY294002和SB203580逆转了上述作用。此外，ADMSCs-Exo抑制了肝纤维化小鼠的病理损伤、纤维化、脂质积累、谷丙转氨酶（ALT）、谷草转氨酶（AST）、总胆红素（TBIL）、CoLI、α-SMA、NOX4、丙二醛（MDA）、PI3K、磷酸化Akt和磷酸化P38MAPK的表达，并提高了白蛋白（ALB）、超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GPx）、过氧化氢酶（CAT）、谷胱甘肽（GSH）、锰超氧化物歧化酶（Mn-SOD）、钠钾ATP酶和钙镁ATP酶的水平。
ADMSCs-Exo通过激活PI3K/Akt/P38MAPK途径抑制氧化应激来减轻肝纤维化，为肝纤维化的潜在治疗提供了有价值的见解。
