藤茶总黄酮对小鼠肝纤维化的改善作用

Improving Effect of Tengeha Flavonoids in Mice with Liver Fibrosis

【目的】探讨藤茶总黄酮(TF)对小鼠肝纤维化的改善作用及机制。【方法】从45只小鼠中随机抽取8只设为正常组,其余37只构建肝纤维化模型。将成功造模的24只小鼠随机分为肝纤维化组、TF组、TF+激活剂组,每组8只。TF组给予300 mg/kg TF灌胃;TF+激活剂组给予300 mg/kg TF、5 mg/kg尼日菌素钠盐灌胃;正常组、肝纤维化组给予等体积生理盐水灌胃。1次/d,连续干预10周。给药结束后,生化检测血清谷丙转氨酶(ALT)、谷草转氨酶(AST)水平,酶联免疫吸附分析(ELISA)检测肝组织白细胞介素18(IL-18)、白细胞介素1β(IL-1β)含量,免疫组织化学染色法检测肝组织α-平滑肌蛋白(α-SAM)表达,马松(Masson)染色法观察肝组织纤维化程度,蛋白免疫印迹(Western Blot)法检测肝组织核苷酸结合寡聚化结构域样受体3(NLRP3)、凋亡相关斑点样蛋白(ASC)、Caspase-1蛋白表达。【结果】与正常组比较,肝纤维化组血清ALT、AST水平,肝组织IL-18、IL-1β含量,α-SAM阳性染色面积百分比,肝组织NLRP3、ASC、Caspase-1蛋白相对表达量升高(P<0.05),Masson染色结果显示,肝纤维化组可观察到大量增生性胶原纤维,肝小叶结构被严重破坏;与肝纤维化组比较,TF组、TF+激活剂组血清ALT、AST水平,肝组织IL-18、IL-1β含量,α-SAM阳性染色面积百分比,肝组织NLRP3、ASC、Caspase-1蛋白相对表达量降低(P<0.05),Masson染色结果显示,TF组、TF+激活剂组肝脏胶原纤维沉积、纤维性增生及假小叶形成有所减少,纤维间隔缩小,而TF组效果优于TF+激活剂组。【结论】TF可改善肝纤维化小鼠肝功能,减轻炎症反应及肝纤维化程度,其机制可能与抑制NLRP3炎症小体通路有关。

Objective To explore the effect of improvement and the mechanism of Tengeha flavonoids(TF)on liver fibrosis in mice.Methods Eight mice were randomly selected from 45 mice as the normal group,and a liver fibrosis model was constructed in the remaining 37 mice.The 24 successfully modeled mice were randomly divided into liver fibrosis group,TF group and TF+activator group,with 8 mice in each group.The TF group was given 300 mg/kg TF by intragastrical gavage;the TF+activator group was given 300 mg/kg TF and 5 mg/kg Nigericin sodium salt by intragastrical gavage;the normal group and liver fibrosis group were given equal volume of normal saline by intragastrical gavage.Intervention was performed once per day for 10 continuous weeks.After administration,serum alanine aminotransferas(ALT)and aspartate aminotransferas(AST)levels were determined by biochemical methods,contents of interleukin 18(IL-18)and interleukin 1β(IL-1β)in liver tissue were measured by enzyme-linked immunosorbent assay(ELISA),α-smooth muscle protein(α-SAM)expression in liver tissue was detected by immunohistochemical staining,the fibrosis degree of liver tissue was observed by Masson staining,and the protein expression of nucleotide-binding oligomerization domain-like receptor 3(NLRP3),apoptosis-associated speck-like protein containing CARD(ASC)and Caspase-1 in liver tissue was detected by Western Blot.Results Compared with the normal group,the levels of serum ALT and AST,the contents of IL-18 and IL-1βin liver tissue,the percentage of positive staining area ofα-SAM,and the relative protein expression levels of NLRP3,ASC and Caspase-1 in liver tissue in liver fibrosis group were increased(P<0.05).Masson staining results showed that,a large number of hyperplastic collagen fibers were seen in the hepatic fibrosis group,and the hepatic lobule structure was severely damaged.Compared with the liver fibrosis group,the levels of serum ALT and AST,the contents of IL-18 and IL-1βin liver tissue,the percentage ofα-SAM positive staining area,and the relative protein expression levels of NLRP3,ASC and Caspase-1 in liver tissue were decreased in TF group and TF+activator group(P<0.05).Masson staining results showed that the liver collagen fiber deposition,fibrous hyperplasia and pseudolobule formation were decreased in TF group and TF+activator group,and fibrous septum was reduced.The effect of TF group was superior to that of TF+activator group.Conclusion TF exerts effect in improving the liver function,reducing the inflammatory response and the degree of liver fibrosis in mice with liver fibrosis,and its mechanism may be related to the inhibition of NLRP3 inflammasome pathway.