垂盆草苷对幼龄肝内胆汁淤积大鼠的作用机制

Mechanism of sarmentosin on juvenile intrahepatic cholestasis in rats

研究垂盆草苷(sarmentosin,SA)对幼龄肝内胆汁淤积大鼠的干预和调节作用。将48只SD幼龄大鼠随机分为正常组(Control)、α-萘异硫氰酸酯(ANIT)组(Model)、熊去氧胆酸(UDCA)阳性对照组、垂盆草苷低、中、高剂量组,每组8只,除正常组外,各组大鼠连续1周分别灌胃给予相应药物,每日1次。在第5 d灌胃给予80 mg/kg ANIT造模。在造模后48 h测定大鼠的胆汁流量;测定大鼠血清中丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、碱性磷酸酶(ALP)的活性和总胆红素(TBIL)、直接胆红素(DBIL)、总胆汁酸(TBA)的含量;检测肝脏组织病理变化和组织匀浆中丙二醛(MDA)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)的含量;测定血清中肿瘤坏死因子(TNF-α)、γ-干扰素(INF-γ)、白细胞介素1β(IL-1β)的表达;Western blot法分析胆汁酸转运蛋白与合成蛋白。与正常组相比,模型组胆汁流量受到明显的抑制;大鼠肝组织出现明显的病理损伤;血清中ALT、AST、ALP、TBIL、DBIL、TBA水平显著升高;组织匀浆中MDA含量显著升高,SOD和GSH-Px含量显著降低;炎症因子TNF-α、IFN-γ、IL-1β的表达升高(P<0.05,P<0.01);FXR、SHP-1、SHP-2、MREP2、BSEP、NTCP蛋白表达下降,CYP7A1、CYP27A1蛋白表达升高。与模型组相比,垂盆草苷各剂量组大鼠的胆汁流量呈现不同程度的增加;大鼠肝脏组织的病理损伤有所改善;血清中ALT、AST、ALP、TBIL、DBIL、TBA水平降低;肝脏组织匀浆中MDA的含量降低,SOD、GSH-Px的含量升高(P<0.05,P<0.01);TNF-α、IFN-γ、IL-1β的表达降低(P<0.05,P<0.01);结果表明垂盆草苷对于胆汁淤积有一定的治疗效果,其中垂盆草苷高剂量的作用效果与UDCA组相当,同时垂盆草苷可上调FXR、SHP-1、SHP-2、MREP2、BSEP、NTCP蛋白表达,下调CYP7A1、CYP27A1蛋白表达,说明垂盆草苷通过调控相关蛋白发挥作用。垂盆草苷对ANIT所致的SD幼龄大鼠肝内胆汁淤积有明显的干预和调节作用。可能是通过参与胆汁酸的转运与合成发挥其治疗作用。

To study the effects of sarmentosin(SA)on the intervention and regulation of juvenile intrahepatic cholestasis in rats,48 young SD rats were randomly divided into the control group,α-naphthylisothiocyanate(ANIT)model group,ursodeoxycholic acid(UDCA)positive control group and low-,medium-and high-dosage groups of SA,with 8 rats in each group.Except for the control group,rats in each group were given corresponding drugs by gavage once a day for a week,and 80 mg/kg ANIT model was established on the 5th day.Bile excretion was measured 48 hours after the establishment of the model;the activities of alanine aminotransferase(ALT),aspartate aminotransferase(AST)and alkaline phosphatase(ALP)and the contents of total bilirubin(TBIL),direct bilirubin(DBIL)and total bile acid(TBA)in serum were measured.The pathological changes of liver tissue and the contents of malondialdehyde(MDA),superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px)in tissue homogenate were detected,and the expressions of tumor necrosis factor-α(TNF-α),γ-interferon(IFN-γ)and interleukin-1β(IL-1β)in serum were determined.Bile acid transporters and synthetic proteins were analyzed by Western blot.Compared with the control group,bile excretion was significantly inhibited in the model group;liver tissue showed obvious pathological damage;serum levels of ALT,AST,ALP,TBIL,DBIL and TBA were significantly increased;MDA content in tissue homogenate was significantly increased,SOD and GSH-Px contents were significantly decreased;inflammatory factors TNF-α,IFN-γand IL-1βwere significantly decreased in the model group.The expression of FXR,SHP-1,SHP-2,MREP2,BSEP and NTCP decreased,while the expression of CYP7A1 and CYP27A1 increased.Compared with the model group,the bile excretion of rats in each dose of SA group increased in varying degrees;the pathological damage of liver tissue was improved;the levels of ALT,AST,ALP,TBIL,DBIL and TBA in serum were decreased;the contents of MDA in liver homogenate were decreased;and the contents of SOD and GSH-Px were increased;the expression of TNF-α,IFN-γand IL-1βdecreased.The results showed that sarmentosin had a certain therapeutic effect on cholestasis.The effect of high dose of SA was similar to that of UDCA group,while SA could up-regulate the expression of FXR,SHP-1,SHP-2,MREP2,BSEP and NTCP,down-regulate the expression of CYP7A1 and CYP27A1,suggesting that the drug plays a role by regulating related proteins.SA has obvious intervention and regulation effect on ANIT-induced intrahe patic cholestasis in young SD rats,with possible therapeutic function by participating in the transport and synthesis of bile acids.