补骨脂炮制前后水煎液致雌性ICR小鼠胆汁瘀积性肝损伤作用比较

Study of cholestatic liver injury in female ICR mice by decoction of Psoraleae Fructus before and after processing

目的探讨补骨脂炮制前后致雌性ICR小鼠胆汁瘀积性肝损伤的差异。方法采用高效液相色谱仪检测补骨脂炮制前后化学成分含量变化。雌性ICR小鼠连续30 d ig生、盐补骨脂水煎液,末次给药24 h后,取血清,采用全自动生化仪检测肝损伤相关指标丙氨酸转氨酶(alanine aminotransferase,ALT)、天冬氨酸转氨酶(aspartate aminotransferase,AST)、碱性磷酸酶(alkaline phosphatase,ALP)活性和总胆红素(total bilirubin,TBIL)、直接胆红素(direct bilirubin,DBIL)、总胆汁酸(total bile acid,TBA)水平;取肝脏、脾脏和肾脏,称定质量并计算脏器指数,采用苏木素-伊红(HE)染色法进行肝脏病理组织学检查;采用ELISA法检测血清中雌二醇(estradiol,E2)、促卵泡激素(follicle-stimulating hormone,FSH)和白细胞介素-6(interleukin-6,IL-6)以及肝脏中法尼醇受体(farnesoid X receptor,FXR)、雌激素硫酸转移酶1E1(sulfotransferase 1E1,SULT1E1)和E2含量。结果补骨脂经盐炙后补骨脂苷、异补骨脂苷、新补骨脂异黄酮含量下降,其余8种成分含量均升高,其中补骨脂素、异补骨脂素、corylifol A升高较为明显。雌性小鼠ig生、盐补骨脂水煎液30 d后出现生殖器损伤,小鼠阴道口红肿,有水样浸润。与对照组比较,生、盐补骨脂组小鼠血清ALT、AST、ALP活性及DBIL、TBIL、TBA水平均显著升高(P<0.05、0.01、0.001),盐炙后补骨脂致肝损伤较生品更严重;肝脏FXR、SULT1E1含量显著降低(P<0.01、0.001),血清E2、FSH、IL-6和肝脏E2含量均显著升高(P<0.05、0.01、0.001)。盐补骨脂与生补骨脂比较,IL-6升高更明显,FXR、SULT1E1下降更明显。结论补骨脂素、异补骨脂素和Corylifol A可能是补骨脂药效成分同时也可能是其主要毒性成分,盐补骨脂较生补骨脂更易造成肝损伤。

Objective To investigate the difference of cholestatic liver injury in female ICR mice induced by raw and salt processed Buguzhi(Psoraleae Fructus).Methods The changes of chemical constituents before and after processing of Psoraleae Fructus were detected by high performance liquid chromatography.Female ICR mice were continuously given raw and salt processed Psoraleae Fructus water decoction for 30 d,and 24 h after last administration,serum was collected,automatic biochemical analyzer was used to detect liver injury-related indicators alanine aminotransferase(ALT),aspartate aminotransferase(AST),alkaline phosphatase(ALP)activities and total bilirubin(TBIL),direct bilirubin(DBIL),total bile acid(TBA)levels.Liver,spleen and kidney were taken and weighed,organ index was calculated;Hematoxylin-eosin(HE)staining was used for liver histopathological examination;ELISA was used to detect contents of estradiol(E2),follicle-stimulating hormone(FSH),interleukin-6(IL-6)in serum and farnesoid X receptor(FXR),estrogen sulfate Transferase 1E1(SULT1E1),E2 in liver.Results After salt processed Psoraleae Fructus,the contents of psoralen,isopsoralen and neobavaisoflavone were decreased,and other eight components contents were increased,among which psoralen,isopsoralen and corylifol A were significantly increased.The genitals of female mice were damaged after ig raw and salt processed Psoraleae Fructus water decoction for 30 d,vaginal mouth of mice was red and swollen with watery infiltration.Compared with control group,ALT,AST,ALP activities and DBIL,TBIL,TBA levels in serum of mice in raw and salt processed Psoraleae Fructus groups were significantly increased(P<0.05,0.01,0.001),liver injury of salt processed Psoraleae Fructus was more serious than that of Psoraleae Fructus.The contents of FXR and SULT1E1 in liver were significantly decreased(P<0.01,0.001),and contents of E2,FSH,IL-6 in serum and E2 in liver were significantly increased(P<0.05,0.01,0.001).Compared with Psoraleae Fructus,IL6 content in salt processed Psoraleae Fructus group was significantly increased,and FXR and SULT1E1 contents were decreased more obviously.Conclusion Psoralen,isopsoralen and Corylifel A may be active components and main toxic components of Psoraleae Fructus,salt processed Psoraleae Fructus is more likely to cause liver damage than raw Psoraleae Fructus.