丹参酮胶囊对α-萘异硫氰酸酯所致肝内胆汁淤积小鼠肝损伤的影响及机制初步研究

Effects of Danshentong Capsules on Intrahepatic Cholestasis Liver Injury in Mice Induced by ANIT

目的探讨丹参酮胶囊对α-萘异硫氰酸酯(α-naphthyl isothiocyanate,ANIT)所致肝内胆汁淤积小鼠肝损伤的影响及其可能机制。方法将C57BL/6小鼠随机分为空白对照组、模型组、丹参酮低剂量组(50 mg·kg^(-1)·d^(-1))、丹参酮中剂量组(125 mg·kg^(-1)·d^(-1))、丹参酮高剂量组(250 mg·kg^(-1)·d^(-1))。检测总胆红素(total bilirubin,TBIL)、碱性磷酸酶(alkaline phosphatase,ALP)、丙氨酸氨基转移酶(alanine aminotransferase,ALT)、天门冬氨酸氨基转移酶(aspartate aminotransferase,AST)的水平,HE染色观察肝脏病理,用LC-MS/MS检测血浆胆汁酸含量,RT-PCR检测孕烯醇酮X受体(pregnenolone X receptor,PXR)、细胞色素P450(cytochrome P450)亚型3A11(CYP3A11)及UDP-葡萄糖醛酸转移酶1A1(UDP-glucuronosyl transferases,UGT1A1)mRNA转录。结果与模型组比,丹参酮低、中剂量组血清ALT、AST和ALP明显下降,肝脏病理有所改善;高剂量组ALT和AST降低不明显,血清TBIL和ALP反而升高,肝脏病理无明显改善。与模型组比较,胆酸(cholic acid,CA)、去氧胆酸(deoxycholic acid,DCA)和石胆酸(lithocholic acid,LCA)在丹参酮低、中剂量组有所减少而高剂量组有所增加。丹参酮增加PXR mRNA转录,并呈剂量依赖关系。CYP3A11和UGT1A1转录在低、中剂量增加倍数明显高于高剂量组。结论丹参酮低剂量对ANIT诱导的肝内胆汁淤积小鼠具有防治作用,高剂量则加重了肝损伤,可能与CYP3A11和UGT1A1在不同剂量间表达差异及其酶活性被抑制有关。

OBJECTIVE To investigate the effect and the relevant mechanism of Danshentong capsules on intrahepatic cholestasis （IC） model mice induced by ANIT. METHODS Female C57BL/6 mouse were randomly divided into normal group, model group, low-dose treatment group （50 mg . kg-1 d-1）, middle-dose treatment group （125 mg kg-1 d-1） and high-dose treatment group （250 mg kg-1 d- 1 ）. Serum TBIL, ALT, AST and ALP were determined and liver pathological changes were observed under micro- scope. The plasma bile acid was determined by LC-MS/MS. Expression of mRNA for PXR, CYP3A11 and UGT1A1 were quantified by RT-PCR. RESULTS Compared with the model group, the levels of ALT, AST and ALP in low-dose group decreased significantly （ P 〈 0. 05 ）, and that in middle-dose group had a downward trend, while TBIL and ALP levels increased significantly in high-dose group （P〈0. 05）. Compared with the model group, concentration of CA, DCA and LCA in plasma in low- and middle-dose group decreased while inereased in high-dose group. Tanshinone could elevate the transcription of mRNA for PXR, CYP3A11 and UGT1A1 （P 〈 0. 05） with dose-dependent mode. CONCLUSION Danshentong Capsules with a lower dosage have protective effect on liver injury for ANIT-indueed IC mice, while that with higher dosage could strength liver injury. The underling mechanism is possible related to the expression difference of CYP3All and UGT1A1 induced by tanshinone with different dosages.