LPS‐induced macrophage exosomes promote the activation of hepatic stellate cells and the intervention study of total astragalus saponins combined with glycyrrhizic acid

Huangqi decoction, also known as Huangqi Liuyi decoction, was first recorded in the prescriptions of the Bureau of Taiping People's Welfare Pharmacy. It comprises astragalus and licorice, which is a commonly used prescription in traditional Chinese medicine for the clinical treatment of chronic liver disease, especially liver cirrhosis. Total astragalus saponins (AST) is the main component of astragalus, and glycyrrhizic acid (GA) is the main component of licorice. In this study, normal macrophage exosomes were extracted, and the exosomes incubated with lipopolysaccharides (LPS) and those incubated with LPS + AST + GA were co-cultured with JS1 cells (hepatic stellate cell line). The survival rate and the activation of key signaling pathways of JS1 cells in each group were detected and compared. We found that the co-culture of LPS-induced macrophage exosomes with JS1 cells could significantly increase the expression levels of Collagen-1 (Col-1) and Alpha smooth muscle actin (α-SMA)in JS1 cells. However, a significant reversal effect was observed after pretreatment with AST combined with GA. Further evaluation found that the expression levels of phospho (p)-Smad2 and p-Smad3 in the JS1 cells were significantly increased after macrophages were induced with LPS, whereas pretreatment with AST + GA could significantly decrease the expression levels of p-Smad2 and p-Smad3. Preliminary results of this study indicated that LPS-induced macrophage exosomes can promote the activation of hepatic stellate cells, and the pretreatment of AST combined with GA can exert a significant intervention effect. In this study, the new mechanism of anti-hepatic fibrosis effect of traditional Chinese medicine components of Huangqi Decoction was analyzed from the perspective of exosomes.摘要:黄芪汤, 又名黄芪六一汤, 始载于《太平惠民和剂局方》, 由黄芪和甘草组成, 是中医临床治疗慢性肝病的常用方剂, 尤其在肝硬化的治疗中有重要作用。黄芪总皂苷(AST)是黄芪的主要成分, 甘草酸(GA)是甘草的主要成分。本研究通过LPS 孵育建立巨噬细胞活化模型, 观察其分泌的外泌体对肝星状细胞 (JS1) 的影响, 进一步采用 AST 联合 GA 预处理巨噬细胞, 观察其干预效应。将各组外泌体分别与 JS1 细胞共培养, 检测各组 JS1 细胞的增殖情况、活化状态及关键信号通路激活情况。研究发现 LPS 诱导的巨噬细胞外泌体可以显著增加 JS1 细胞Col1、α-SMA 表达水平, 促进 JS1 细胞的活化。然而, 对巨噬细胞进行 AST 联合 GA 的预处理后具有显著的逆转作用。同时, LPS 诱导巨噬细胞后 JS1 细胞的 p-Smad2、p-Smad3 表达水平显着升高, 而 AST 联合 GA 预处理可降低 p-Smad2、 p-Smad3 的表达。本研究结果表明:脂多糖诱导的巨噬细胞外泌体可以促进肝星状细胞活化, AST 联合 GA 预处理能发挥显著的干预效应。本研究从外泌体角度解析了黄芪汤中药组分抗肝纤维化的新机制。.