薯蓣皂苷对硫代乙酰胺诱导急性肝损伤模型大鼠的保护作用

Protective Effect of Diosgenin on Thioacetamide-Induced Acute Liver Injury in Model Rats

目的探讨薯蓣皂苷通过法尼酯衍生物X受体(FXR)/AMP活化蛋白激酶(AMPK)信号通路对硫代乙酰胺(TAA)诱导急性肝损伤模型大鼠的保护作用。方法将60只雄性大鼠均分为正常对照组(A组),模型组(B组),薯蓣皂苷低剂量组(C_(1)组)、中剂量组(C_(2)组)、高剂量组(C_(3)组)和阳性对照组(D组),各10只。C_(1)组、C_(2)组、C_(3)组大鼠分别灌胃薯蓣皂苷25,50,100 mg/kg,D组大鼠灌胃水飞蓟素200 mg/kg,A组和B组大鼠灌胃生理盐水每100 g体质量0.1 mL,每天1次,连续7 d。末次灌胃2 h后,B组、C_(1)组、C_(2)组、C_(3)组、D组大鼠腹腔注射TAA 100 mg/kg构建急性肝损伤模型,A组注射等量生理盐水。禁食不禁饮,24 h后麻醉大鼠,心脏取血,采用自动生物分析仪测定血清中丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)的水平,采用酶联免疫吸附(ELISA)法测定血清中肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)6、IL-10的水平,以及肝脏组织匀浆中谷胱甘肽(GSH)、超氧化物歧化酶(SOD)的活性和丙二醛(MDA)的含量,采用蛋白免疫印迹法测定肝组织中FXR、AMPK、核因子E2相关因子2(Nrf2)、核转录因子κB(NF-κB)的蛋白表达水平。结果与A组比较,B组、C_(1)组、C_(2)组、C_(3)组、D组大鼠ALT,AST,TNF-α,IL-6,MDA水平及NF-κB蛋白表达水平均显著升高,IL-10,GSH,SOD水平,以及FXR,AMPK,Nrf2蛋白表达水平均显著降低(P<0.05);与B组比较,C1组、C2组、C3组、D组大鼠ALT,AST,TNF-α,IL-6,MDA水平及NF-κB蛋白表达水平均显著降低,IL-10,GSH,SOD水平,以及FXR,AMPK,Nrf2蛋白表达水平均显著升高,且C_(1)组、C_(2)组、C_(3)组呈剂量依赖性(P<0.05);D组和C3组各指标比较无显著差异(P>0.05)。结论薯蓣皂苷对TAA诱导的急性肝损伤模型大鼠具有保护作用,其作用机制可能与激活FXR/AMPK信号通路有关。

Objective To investigate the protective effect of diosgenin on thioacetamide(TAA)-induced acute liver injury in model rats through farnesoid X receptor(FXR)/AMP-activated protein kinase(AMPK)signaling pathway.Methods A total of 60 male rats were divided into the normal control group(group A),model group(group B),diosgenin low-dose group(group C1),diosgenin medium-dose group(group C2),diosgenin high-dose group(group C3)and positive control group(group D),10 rats in each group.The rats in groups C1,C2and C3 were given 25,50,100 mg/kg of diosgenin by gavage,the rats in group D were given 200 mg/kg of silymarin by gavage,,and the rats in groups A and B were give 0.1 m L of normal saline per 100 g,once a day for 7 d.Two hours after the last intragastric administration,the rats in groups B,C1,C2,C3and D were injected with 100 mg/kg of TAA intraperitoneally to establish the rat model of acute liver injury,and the rats in group A were injected with equivalent normal saline.After fasting without forbidding drink for 24 h,they were anesthetized and the blood was removed from the heart.The levels of serum alanine aminotransferase(ALT)and aspartate aminotransferase(AST)were measured by the automatic biochemical analyzer,the levels of serum tumor necrosis factor-α(TNF-α),interleukin(IL)-6 and IL-10,the activity of glutathione(GSH),superoxide dismutase(SOD),and the content of malondialde-hyde(MDA)in liver homogenate were determined by the enzyme-linked immunosorbent assay(ELISA),and the expression levels of FXR,AMPK,nuclear factor E2 related factor 2(Nrf2)and nuclear transcription factor-κB(NF-κB)protein in liver tissue were mea-sured by Western blot.Results Compared with those in group A,the levels of ALT,AST,TNF-α,IL-6,MDA and the expression level of NF-κB protein in groups B,C1,C2,C3and D increased significantly,while the levels of IL-10,GSH,SOD,and the expression levels of FXR,AMPK and Nrf2 protein in groups B,C1,C2,C3and D decreased significantly(P<0.05).Compared with those in group B,the levels of ALT,AST,TNF-α,IL-6,MDA and the expression level of NF-κB protein in groups C1,C2,C3and D decreased signifi-cantly,while the levels of IL-10,GSH,SOD,and the expression levels of FXR,AMPK and Nrf2 protein increased significantly,and the effect in the groups C1,C2and C3 were dose-dependent(P<0.05).There was no significant difference in the above indexes between group D and group C3(P>0.05).Conclusion Diosgenin has a protective effect on TAA-induced acute liver injury in model rats,and its mechanism may be related to the activation of the FXR/AMPK signaling pathway.