金钗石斛生物碱对四氯化碳诱导急性肝损伤小鼠的作用

Effects of alkaloids of Dendrobium nobile Lindl. on acute liver injury induced by carbon tetrachloride in mice

目的观察金钗石斛生物碱(ADNL)对四氯化碳(CCl4)诱导急性肝损伤小鼠的作用。方法50只C57小鼠随机分为正常对照组,正常给药(ADNL 20 mg·kg^(-1)·d^(-1))组,模型组和ADNL低、高剂量(10、 20 mg·kg^(-1)·d^(-1))组,每组10只,连续灌胃7 d。末次给药后2 h,除正常对照组和正常给药组外,其余各组腹腔注射0.2%CCl4玉米油溶液(10 mL·kg^(-1))建立急性肝损伤模型。采用全自动生化仪检测小鼠血清中丙氨酸转氨酶(ALT)和天冬氨酸转氨酶(AST)含量, HE染色观察肝组织病理学变化, RT-PCR检测肝组织中肿瘤坏死因子(TNF)-α、白细胞介素1β(IL-1β)和白细胞介素6 (IL-6) mRNA表达水平, Western blot检测TNF-α、 IL-6、 p-核转录因子(NF)-κB p65蛋白表达水平。结果与正常对照组比较,正常给药组ALT、 AST及肝组织病理学无明显变化,模型组小鼠血清ALT、 AST的含量均明显升高(P <0.05),肝脏组织可见明显病理损伤, TNF-α、 IL-1β和IL-6 mRNA和IL-6、 TNF-α和p-NF-κB p65蛋白表达显著增加(P <0.05)。与模型组比较, ADNL低、高剂量组血清ALT和AST含量显著降低(P <0.05),肝组织病理损伤明显改善, TNF-α和IL-1βmRNA表达显著下调(P <0.05), TNF-α和p-NF-κB p65蛋白表达显著减少(P <0.05), ADNL高剂量组IL-6蛋白和mRNA表达也显著下调(P <0.05)。结论 ADNL对CCl4所致急性肝损伤具有显著改善作用,可能与抑制炎症反应有关。

AIM To investigate the effects of alkaloids of Dendrobium nobile Lindl.(ADNL)on acute liver injury induced by carbon tetrachloride(CCl4)in mice.METHODS Fifty C57 mice were randomly divided into normal control group,normal administration(ADNL 20 mg·kg-1·d-1)group,model group,ADNL low dose(10 mg·kg-1·d-1)and high dose(20 mg·kg-1·d-1)group(n=10).Mice were all administered intragastrically for 7 days.Except the normal control group and normal administration group,the other groups were intraperitoneally injected with 0.2%CCl4 corn oil solution(10 mL·kg-1)to induce acute liver injury.The contents of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)in serum were detected by automatic biochemical analyzer.The pathological changes of liver tissue were assessed by HE staining.The m RNA expression of tumor necrosis factor(TNF)-α,interleukin 1β(IL-1β)and interleukin 6(IL-6)were detected by RT-PCR.And the protein expression of TNF-α,IL-6 and phosphorylation-nuclear factorκB p65(p-NF-κB p65)in liver tissue were analyzed by Western blot.RESULTS Compared with the normal control group,there were no significant changes in ALT,AST and pathological tissues in the normal administration group.The serum levels of ALT and AST in the model group were significantly increased(P<0.05),and the pathological damage was seen in the liver tissue apparently,and the mRNA expression of TNF-α,IL-1βand IL-6 and the protein expression of IL-6,TNF-αand p-NF-κB p65 were also increased significantly(P<0.05).Compared with the model group,the contents of ALT and AST were reduced significantly in the ADNL low and high dose groups and the pathological damage of liver tissue were improved.At the same time,the m RNA expression of TNF-αand IL-1βand the protein expression of TNF-αand p-NF-κB p65 were down-regulated(P<0.05).And the mRNA and protein expression of IL-6 were down-regulated in the ADNL low dose group(P<0.05).CONCLUSION ADNL has a significant improvement effects on acute liver injury induced by CCl4,which may be related to inhibition of inflammatory response.