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降黄合剂Ⅱ号对肝内胆汁淤积大鼠肝损伤及肝组织中NTCP和BSEP表达的影响 被引量:2

Effects of Jianghuang mixtureⅡon liver injury and expression of NTCP and BSEP in rats with intrahepatic cholestasis
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摘要 目的探讨降黄合剂Ⅱ号对复合因素诱导的阴黄证肝内胆汁淤积模型大鼠肝损伤及钠-牛黄胆酸盐共转运多肽(NTCP)、胆盐输出泵(BSEP)表达的影响。方法将60只SD雄性大鼠随机分为正常组、模型组、降黄合剂Ⅱ号高剂量组、降黄合剂Ⅱ号中剂量组、降黄合剂Ⅱ号低剂量组,每组12只。正常组大鼠予常规饲养;其余各组大鼠造模阶段施以寒湿条件、大黄煎剂灌胃及高糖高脂饮食,造模3周,第22天时模型组及降黄合剂Ⅱ号各组给予0.5%的ANIT 20 mg/100 g灌胃1次,正常组予以相同体积生理盐水灌胃,建立阴黄证肝内胆汁淤积大鼠模型。降黄合剂Ⅱ号各组于造模第8天开始每天下午给相应剂量的降黄合剂Ⅱ号灌胃,一直到造模后14 d,共用药28 d,正常组和模型组均予相同剂量生理盐水灌胃。观察各组大鼠一般状态,检测干预结束后各组大鼠血清碱性磷酸酶(ALP)、谷丙转氨酶(ALT)、谷草转氨酶(AST)、总胆红素(TBil)水平,镜下观察各组大鼠肝组织病理变化,采用Western blot法检测各组大鼠肝组织中BSEP、NTCP蛋白表达水平。结果正常组大鼠饮食量、活动量、被毛、体质量、排便排尿情况均正常;模型组大鼠形体均较消瘦,饮食量减少,大便增多,尿液明显变黄、增多,被毛枯黄、粗糙且易脱毛,精神状态不佳;降黄合剂Ⅱ号各组大鼠干预后情况均较模型组明显好转,以大剂量组最为明显。模型组大鼠血清ALP、ALT、AST、TBil水平均明显高于正常组(P均<0.05);降黄合剂Ⅱ号大剂量组和降黄合剂Ⅱ号中剂量组上述指标水平均明显低于模型组(P均<0.05),且降黄合剂Ⅱ号大剂量组各指标水平均明显低于降黄合剂Ⅱ号小剂量组(P均<0.05),其中血清ALP、ALT、TBil水平明显低于降黄合剂Ⅱ号中剂量组(P均<0.05)。HE染色显示降黄合剂Ⅱ号大、中剂量组肝组织较模型组中炎细胞浸润减少,毛细胆管淤堵减轻,肝细胞肿胀缓解,细胞核清晰,形态较完整,可见少量炎性细胞。模型组大鼠肝组织中NTCP、BSEP相对表达量均明显低于正常组(P均<0.05);降黄合剂Ⅱ号大剂量组和降黄合剂Ⅱ号中剂量组大鼠肝组织中NTCP、BSEP相对表达量均明显高于模型组(P均<0.05),降黄合剂Ⅱ号小剂量组与模型组比较差异均无统计学意义(P均>0.05)。结论降黄合剂Ⅱ号可改善阴黄证肝内胆汁淤积模型大鼠肝损伤,机制可能与上调NTCP和BSEP表达有关。 Objective It is to investigate the effects of Jianghuang MixtureⅡon liver injury and the expression of Na+/taurocholate cotransporting polypeptide(NTCP)and bile salt export pump(BSEP)in rats with intrahepatic cholestasis of Yinhuang syndrome induced by multiple factors.Methods Sixty male SD rats were randomly divided into normal group,model group,Jianghuang mixture No.Ⅱhigh-dose group,Jianghuang mixture No.Ⅱmiddle-dose group,and Jianghuang mixture No.Ⅱlow-dose group,12 rats in each group.The rats in the normal group were routinely fed;the rats in the other groups were treated with rhubarb decoction by gavage and high-sugar and high-fat diet for 3 weeks under cold and wet conditions.On the 22nd day,the model group and each Jianghuang mixture No.Ⅱgroup was intragastrically administered with 0.5 mg of ANIT 20 mg/100 g,and the normal group was intragastrically administered with the same volume of normal saline to establish a rat model of intrahepatic cholestasis of Yinhuang syndrome.Each group of Jianghuang Mixture No.Ⅱwas administrated with the corresponding dose of Jianghuang Mixture No.Ⅱevery day starting from the 8th day of modeling,until 14 days after modeling,and the medicine was used for 28 days.The normal group and model group were given the same dose of normal saline by gavage.The general status of the rats in each group was observed,and the levels of serum alkaline phosphatase(ALP),alanine aminotransferase(ALT),aspartate aminotransferase(AST),and total bilirubin(TBil)in each group after the intervention were detected,the pathological changes of liver tissue of the rats in each group were observed under microscope.Western blot was used to detect the expression levels of BSEP and NTCP proteins in liver tissue of rats in each group.Results The diet,activity,coat,body weight,and defecation was normal in the rats of normal group.The rats in the model group were thinner,with reduced diet,increased stools,markedly yellowing and increasing urine,yellowish coat,rough and easy hair removal,and poor mental state.After intervention,the status of the rats in each group of Jianghuang Mixture No.Ⅱwas improved significantly compared with the model group,especially in the high-dose group.The levels of serum ALP,ALT,AST,and TBil in the model group were significantly higher than those in the normal group(all P<0.05);the levels of the above indicators were significantly lower in the Jianghuang MixtureⅡhigh-dose group and the Jianghuang MixtureⅡmiddle-dose group than those in the model group(all P<0.05),and the levels of these indicators in the Jianghuang mixture No.Ⅱhigh-dose group were significantly lower than those of the Jianghuang mixture No.Ⅱlow-dose group(P<0.05),in which the levels of serum ALP,ALT,and TBil were significantly lower than those in the middle-dose group of Jianghuang Mixture No.Ⅱ(P<0.05).HE staining showed that the liver tissue of Jianghuang Mixture No.Ⅱhigh and medium dose groups had less infiltration of inflammatory cells,less choledocholithiasis,less hepatocellular swelling,clearer nuclei,more complete morphology,and a small number of inflammatory cells.The relative expressions of NTCP and BSEP in liver tissue of model group rats were significantly lower than those in normal group(both P<0.05);NTCP,The relative expression levels of BSEP were significantly higher than those of the model group(all P<0.05),and there was no significant difference between the low-dose Jianghuang mixtureⅡgroup and the model group(all P>0.05).Conclusion Jianghuang Mixture No.Ⅱcan improve liver injury in the model of Yinhuang syndrome intrahepatic cholestasis in rats.The mechanism may be related to the up-regulation of NTCP and BSEP expression.
作者 席奇 杨璇 李京涛 闫曙光 魏海梁 李日向 常占杰 XI Qi;YANG Xuan;LI Jingtao;YAN Shuguang;WEI Hailiang;LI Rixiang;CHANG Zhanjie(Affiliated Hospital of Shaanxi University of Traditional Chinese Medicine,Xianyang 712000,Shaanxi,China;Shaanxi University of Traditional Chinese Medicine,Xianyang 712000,Shaanxi,China)
出处 《现代中西医结合杂志》 CAS 2020年第9期913-917,944,共6页 Modern Journal of Integrated Traditional Chinese and Western Medicine
基金 陕西中医药大学青年基金项目(2016QN13) 陕西中医药大学学科创新团队建设项目(2019-YL05)。
关键词 降黄合剂Ⅱ号 肝内胆汁淤积 钠-牛黄胆酸盐共转运多肽 胆盐输出泵 Jianghuang MixtureⅡ intrahepatic cholestasis NTCP BSEP
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