The aim of this study was to investigate the underlying protective mechanisms of asiaticoside (AS) against liver fibrosis (LF) both in vivo and in vitro. A rat model with carbon tetrachloride (CCl₄)-induced liver fibrosis is employed to verify the effect and mechanism of AS on the process of liver fibrosis in vivo experiment. Hematoxylin/eosin and sirius red staining was conducted to assess the severity of liver injury and fibrosis. Further, the serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), albumin (ALB), glutamyl transferase (GGT), and total bilirubin (TBil) were measured. In addition, LX2 cells were cultured for vitro experiment to investigate the influence of AS on hepatic stellate cells (HSCs). Overproduction of α-smooth muscle actin and type I collagen is characteristic of LF and HSCs, as determined by immunohistochemical and Western blot analyses. The expression levels of molecules associated with the Notch signaling pathway (i.e., Notch-1, Jagged-1, and Delta-like-4) were assessed by Western blot analysis. The results revealed that AS attenuated LF, as defined by reduced deposition of collagen, expression of α-smooth muscle actin and collagen type 1, and expression of biochemical parameters (alanine aminotransferase, aspartate aminotransferase, and hydroxyproline). Notably, AS suppressed the expression levels of Notch-1, Jagged-1, and Delta-like-4 in activated HSCs and LF. Collectively, these results demonstrate that AS prevented the progression of LF by modulating the Notch signaling pathway, indicating that AS has potential therapeutic effects against LF.

本研究的目的是研究积雪草苷 (AS) 在体内和体外对肝纤维化 (LF) 的潜在保护机制。四氯化碳（CCl ₄)-诱导肝纤维化用于体内实验验证AS对肝纤维化过程的影响和机制。进行苏木精/伊红和天狼星红染色以评估肝损伤和纤维化的严重程度。此外，测量了丙氨酸氨基转移酶（ALT）、天冬氨酸氨基转移酶（AST）、白蛋白（ALB）、谷氨酰转移酶（GGT）和总胆红素（TBil）的血清水平。此外，培养LX2细胞进行体外实验，研究AS对肝星状细胞（HSCs）的影响。α-平滑肌肌动蛋白和 I 型胶原蛋白的过量产生是 LF 和 HSC 的特征，如通过免疫组织化学和蛋白质印迹分析所确定的。Notch信号通路相关分子的表达水平（即Notch-1、Jagged-1、和 Delta-like-4) 通过蛋白质印迹分析进行评估。结果显示 AS 减弱了 LF，定义为胶原蛋白沉积减少、α-平滑肌肌动蛋白和 1 型胶原蛋白的表达以及生化参数（谷丙转氨酶、天冬氨酸转氨酶和羟脯氨酸）的表达。值得注意的是，AS 抑制了激活的 HSC 和 LF 中 Notch-1、Jagged-1 和 Delta-like-4 的表达水平。总的来说，这些结果表明 AS 通过调节 Notch 信号通路阻止了 LF 的进展，表明 AS 对 LF 具有潜在的治疗作用。天冬氨酸氨基转移酶和羟脯氨酸）。值得注意的是，AS 抑制了激活的 HSC 和 LF 中 Notch-1、Jagged-1 和 Delta-like-4 的表达水平。总的来说，这些结果表明 AS 通过调节 Notch 信号通路阻止了 LF 的进展，表明 AS 对 LF 具有潜在的治疗作用。天冬氨酸氨基转移酶和羟脯氨酸）。值得注意的是，AS 抑制了激活的 HSC 和 LF 中 Notch-1、Jagged-1 和 Delta-like-4 的表达水平。总的来说，这些结果表明 AS 通过调节 Notch 信号通路阻止了 LF 的进展，表明 AS 对 LF 具有潜在的治疗作用。