摘要
目的:探索柴胡-白芍配伍对利福平(RFP)诱导的胆汁淤积大鼠的治疗作用及其机制。方法:60只雄性SD大鼠随机分为正常组、模型组、熊去氧胆酸(UDCA)组、柴胡组、白芍组、柴-白组,每组10只,上午除正常组外大鼠采用RFP混悬液灌胃复制胆汁淤积模型,下午各组灌胃相应生理盐水或药物治疗,持续14 d。检测血清丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)、总胆红素(TBil)、直接胆红素(DBil)、碱性磷酸酶(ALP)、总胆汁酸(TBA)含量,检测肝脏组织TBA含量,HE染色观察肝脏病理组织形态学变化,qRT-PCR检测肝组织胆盐输出泵(BSEP)、多重耐药相关蛋白2(MRP2)、多重耐药蛋白3(MDR3)、闭锁小带蛋白-1(ZO-1)、连接黏附分子-A(JAM-A)、闭合蛋白(Occludin)、连接蛋白(Claudin)1、Claudin2、Claudin3基因表达水平,Western blot检测肝组织BSEP、MRP2、MDR3、ZO-1、Occludin蛋白表达量。结果:RFP诱导的胆汁淤积大鼠食欲、精神欠佳,毛发蓬乱且黄染严重,皮毛无光泽出现掉毛现象,脚掌、尾部及大小便均呈橘黄色改变,体质量增长较缓慢;肝湿质量比值、肝组织TBA含量及各肝功能指标ALT、AST、ALP、TBil、DBil、TBA含量明显升高(P<0.01);HE染色结果显示肝细胞小叶结构较完整,出现部分炎性浸润、肝细胞水肿,散在肝细胞脂肪空泡及部分胆汁淤积;肝组织BSEP、MRP2、MDR3、ZO-1、Occludin基因及蛋白表达明显降低(P<0.01),JAM-A、Claudin1、Claudin2和Claudin3基因表达略有下降,但差异无统计学意义(P>0.05)。药物治疗后,各项指标显示出不同程度的好转。结论:柴胡-白芍配伍可通过影响肝细胞极性改善RFP诱导的大鼠胆汁淤积。
Objective:To investigate the therapeutic mechanism of Bupleurum-Paeonia compatibility in treating cholestasis induced by rifampicin(RFP).Methods:Sixty male Sprague-Dawley(SD)rats were randomly divided into six groups(n=10 per group):a normal control group,a model group,a ursodeoxycholic acid(UDCA)group,a Bupleurum group,a Paeonia group,and a Bupleurum-Paeonia combination group.Except for the normal control group,rats in all other groups were administered RFP suspension by oral gavage in the morning to establish a cholestasis model.In the afternoon,rats in each group received intragastric administration of either physiological saline or the corresponding therapeutic agents.The treatment lasted for 14 consecutive days.Serum levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),total bilirubin(TBil),direct bilirubin(DBil),alkaline phosphatase(ALP),and total bile acids(TBA)were measured.Hepatic TBA content was also determined.Liver histopathological changes were observed by hematoxylin-eosin(HE)staining.The mRNA expression levels of bile salt export pump(BSEP),multidrug resistance-associated protein 2(MRP2),multidrug resistance protein 3(MDR3),zonula occludens-1(ZO-1),junctional adhesion molecule-A(JAM-A),occludin,claudin-1,claudin-2,and claudin-3 in liver tissues were assessed using quantitative real-time PCR(qRT-PCR).Protein expression levels of BSEP,MRP2,MDR3,ZO-1,and occludin were analyzed by Western blotting.Results:Rats with RFP-induced cholestasis exhibited reduced appetite and activity,rough and yellow-stained fur with hair loss,dull coat appearance,orange discoloration of the paws,tail,urine,and feces,as well as slower body-weight gain.The liver wet weight ratio,hepatic TBA content,and serum liver function indicators-including ALT,AST,ALP,TBil,DBil,and TBA-were significantly increased compared with the normal control group(P<0.01).HE staining revealed relatively preserved hepatic lobular architecture,accompanied by partial inflammatory cell infiltration,hepatocellular edema,scattered lipid vacuoles,and focal bile stasis.The mRNA and protein expression levels of BSEP,MRP2,MDR3,ZO-1,and occludin in liver tissues were markedly decreased(P<0.01).Although the mRNA expression levels of JAM-A,claudin-1,claudin-2,and claudin-3 showed a slight downward trend,the differences were not statistically significant(P>0.05).Following drug treatment,varying degrees of improvement were observed in all measured indicators.Conclusion:The combination of Bupleurum and Paeonia alleviates RFP-induced cholestasis in rats by modulating hepatocyte polarity.
作者
张梦杰
王雅静
曹娜娜
刘云帆
王茜
ZHANG Meng-jie;WANG Ya-jing;CAO Na-na;WANG Xi(School of Pharmacy,Hebei University of Chinese Medicine(Shijiazhuang Hebei,050091),China)
出处
《中西医结合肝病杂志》
2025年第12期1492-1499,共8页
Chinese Journal of Integrated Traditional and Western Medicine on Liver Diseases
基金
河北省自然基金中医药联合基金(No.H2022423305)。
关键词
胆汁淤积
利福平
柴胡
白芍
配伍
肝细胞极性
cholestasis
rifampicin
Bupleurum
Paeonia
compatibility
hepatocyte polarity
