茵陈蒿汤对自身免疫性肝炎小鼠肝细胞铁死亡的抑制作用 及其机制分析.

Role and mechanism of action of Yinchenhao Decoction in inhibiting ferroptosis of hepatocytes in mice with autoimmune hepatitis.

Objective To investigate the role and mechanism of action of Yinchenhao Decoction in inhibiting ferroptosis of hepatocytes in mice with autoimmune hepatitis. Methods A total of 18 specific pathogen-free female C57BL/6 mice were selected and divided into normal group, model group, and treatment group using a random number table, with 6 mice in each group. The mice in the model group and the treatment group were injected with concanavalin A (Con A) via the caudal vein to establish a mouse model of autoimmune hepatitis, and those in the normal group were injected with normal saline. The mice in the treatment group were given prophylactic treatment with Yinchenhao Decoction (4.68 g crude drug/kg) by gavage at 14 days before modeling, and Con A was injected after the last gavage. The levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), interferon gamma (IFN-γ), tumor necrosis factor-α (TNF-α), iron ion, glutathione (GSH), reactive oxygen species (ROS), adenosine triphosphate (ATP), and malondialdehyde (MDA) were measured； liver index and spleen index were calculated； the expression levels of GPX4 and SLC7A11 were measured； liver histopathological changes were compared between groups. A oneway analysis of variance was used for comparison of normally distributed continuous data between three groups, and the least significant difference t-test was used for further comparison between two groups. Results Compared with the normal group, the model group had significant increases in liver index, spleen index, ALT, AST, IFN-γ, TNF-α, iron ion, ROS and MDA (all P< 0.05) and significant reductions in the content of GSH and ATP and the protein expression levels of GPX4 and SLC7A11 (all P< 0.05). Compared with the model group, the treatment group had significant reductions in liver index, spleen index, ALT, AST, IFN-γ, TNF-α, iron ion, ROS and MDA (all P<0.05) and significant increases in the content of GSH and ATP and the protein expression levels of GPX4 and SLC7A11 (all P<0.05). HE staining showed that compared with the normal group, the model group showed massive hepatocyte degeneration and necrosis and inflammatory cell aggregation at the portal area, and compared with the model group, the treatment group had alleviation of liver necrosis and inflammatory infiltration. Conclusion Liver injury induced by Con A may be associated with ferroptosis. Yinchenhao Decoction can increase the protein expression levels of SLC7A11 and GPX4 protein and thus inhibit ferroptosis of hepatocytes induced by Con A. [ABSTRACT FROM AUTHOR]

目的　探讨茵陈蒿汤对自身免疫性肝炎小鼠肝细胞铁死亡的抑制作用及机制。方法　选取 SPF级雌性 C57BL/6小 鼠18只，采用随机数字表法分为正常组, 模型组, 治疗组，每组6只。模型组和治疗组采用刀豆蛋白A (Con A) 尾静脉注射制 备自身免疫性肝炎小鼠模型，正常组注射生理盐水。治疗组于造模前 14天，给予茵陈蒿汤 (4. 68 g生药/kg) 预防性灌胃治 疗，末次灌胃给药后注射Con A。分别检测ALT, AST, IFN-γ, TNF-α, 铁离子, GSH, ROS, ATP, MDA水平，计算肝脏指数, 脾 脏指数，并观察GPX4, SLC7A11的表达；比较各组小鼠肝脏病理组织变化。正态分布的计量资料3组间比较采用单因素方 差分析，进一步两两比较采用LSD-t检验。结果　与正常组比较，模型组肝脏指数, 脾脏指数, ALT, AST, IFN-γ, TNF-α, 铁离 子, ROS, MDA 水平均升高 (P值均<0. 05) ，GSH, ATP含量及 GPX4, SLC7A11蛋白表达水平降低 (P值均<0. 05) 。与模型组 比较，治疗组肝脏指数, 脾脏指数, ALT, AST, IFN-γ, TNF-α, 铁离子, ROS, MDA 水平均降低 (P值均<0. 05) ，GSH, ATP含量 及GPX4, SLC7A11蛋白表达水平升高 (P值均<0. 01) 。HE染色结果显示，与正常组比较，模型组小鼠肝脏出现大面积肝细 胞变性, 坏死, 门管区炎细胞聚集，治疗组处理的小鼠肝脏坏死和炎性浸润程度较模型组有所减轻。结论 Con A诱导肝组 织损伤可能与铁死亡有关。茵陈蒿汤可以增加SLC7A11/GPX4蛋白的表达水平进而抵抗Con A诱导的肝细胞铁死亡。 [ABSTRACT FROM AUTHOR]