摘要
目的 探讨五灵胶囊对自身免疫性肝炎(AIH)小鼠的保护作用.方法 小鼠随机分为对照组、AIH模型组、五灵胶囊低剂量组(0.5 g·kg-1·d-1)、五灵胶囊中剂量组(1.0 g·kg-1·d-1)、五灵胶囊高剂量组(2.0 g·kg-1·d-1),每组10只.五灵胶囊各剂量组按10 mL/kg的体积灌胃给予五灵胶囊混悬液,每天1次;对照组和AIH模型组灌胃给予等体积生理盐水.给药14 d后,AIH模型组和五灵胶囊各剂量组小鼠尾静脉注射刀豆蛋白A(20 mg/kg),注射 8 h后收集血清、肝脏和脾脏.采用全自动生化分析仪检测血清丙氨酸转氨酶(ALT)和天冬氨酸转氨酶(AST)水平,用H-E染色观察肝组织病理结构,用ELISA法测定肝组织中IL-6、IL-4 和TNF-α含量,用qPCR法测定肝组织中IL-6、IL-4、TNF-α、Toll样受体4(TLR4)和NF-κB mRNA的相对表达量,用免疫荧光法分析肝组织中NF-κB p65 蛋白表达和核移位情况.结果 与对照组相比,AIH模型组小鼠肝脏形态结构异常,血清ALT和AST水平升高,肝组织中IL-4、IL-6 和TNF-α含量升高,肝组织中IL-6、IL-4、TNF-α、TLR4和NF-κB mRNA的相对表达量上调,NF-κB p65 的入核增加.五灵胶囊明显改善AIH模型小鼠的肝脏病理结构,降低血清ALT和AST水平,降低肝组织中IL-4、IL-6 和TNF-α含量及TLR4、NF-κB、IL-4、IL-6 mRNA的相对表达量,并抑制NF-κB p65的入核激活.结论 五灵胶囊对AIH小鼠具有明显的保护作用,其机制可能与TLR4/NF-κB信号通路有关.
Abstract
Objective To investigate the protective effects of Wuling capsule on mice with autoimmune hepatitis(AIH).Methods Mice were randomly divided into control group,AIH model group,Wuling capsule low-dose group(0.5 g·kg-1·d-1),Wuling capsule middle-dose group(1.0 g·kg-1·d-1),and Wuling capsule high-dose group(2.0 g·kg-1·d-1),with 10 mice in each group.The Wuling capsule groups were administered with Wuling capsule suspension of different doses orally at a volume of 10 mL/kg once daily;the control group and AIH model group were given the same volume of saline by gavage.After 14 d of administration,mice in the AIH model group and Wuling capsule groups were injected with concanavalin A(20 mg/kg)via the tail vein,and the serum,liver,and spleen were collected 8 h after injection.Serum alanine transaminase(ALT)and aspartate transaminase(AST)levels were measured using an automatic biochemical analyzer;hematoxylin-eosin staining was used to observe the pathological structure of the liver tissue;the contents of interleukin(IL)-6,IL-4,and tumor necrosis factor(TNF)-α in liver were determined by enzyme-linked immunosorbent assay,and quantitative polymerase chain reaction was used to determine the relative mRNA expression levels of IL-6,IL-4,TNF-α,Toll-like receptor 4(TLR4),and nuclear factor κB(NF-κB)in the liver.Fluorescence immunoassay was used to analyze the expression and nuclear translocation of NF-κB p65 protein in the liver.Results Compared with the control group,the AIH model group showed abnormal liver morphology and structure,increased serum ALT and AST levels,increased contents of IL-4,IL-6,and TNF-α in the liver,upregulated mRNA expression levels of IL-6,IL-4,TNF-α,TLR4,and NF-κB in the liver,and increased nuclear entry of NF-κB p65.Wuling capsule significantly improved the pathological structure of the liver in AIH mice,reduced serum ALT and AST levels,decreased the contents of IL-4,IL-6,and TNF-α in the liver and the mRNA expression levels of TLR4,NF-κB,IL-4,IL-6,and inhibited the nuclear activation of NF-κB p65.Conclusion Wuling capsule has significant protective effects on AIH mice,which may be related to the TLR4/NF-κB signaling pathway.
维普
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