基于转运体的何首乌致肝损伤作用机制探讨

目的：阐明长期给药何首乌醇提物 (PME) 和水提物 (PMW) 致大鼠肝损伤的可能机制。方法：分别制备 PME 和 PMW。雄性 SD 大鼠随机分为对照组、阳性对照异硫氰酸 α-萘酯 (ANIT 100 mg·kg^–1) 组、PMW组 (15 g·kg^–1)、PME 组 (12 g·kg^–1)。PME、PMW 组大鼠给药剂量相当于 60 g·kg^–1生药量，约为临床最大量的110 倍，灌胃给药 42 d，每日给药 1 次。ANIT 组于第 40 天灌胃给药 1 次。采用试剂盒方法检测大鼠血清生化指标丙氨酸氨基转移酶 (ALT)、天冬氨酸氨基转移酶 (AST)、总胆汁酸 (TBA)、碱性磷酸酶 (ALP)、总胆红素(TBIL) 水平；采用苏木素-伊红 (HE) 染色法检测大鼠肝脏病理变化；采用实时荧光定量聚合酶链式反应 (qRT-PCR) 测 定 给 药 后 大 鼠 肝 脏 组 织 中 胆 汁 酸 、 胆 红 素 代 谢 相 关 转 运 体 有 机 阴 离 子 转 运 多 肽 1a1 (OATP1a...

Transporter-based Mechanism of Liver Injury Caused by Polygoni Multiflori Radix

Objective To decipher the mechanism of rat liver injury induced by long-term administration of Polygoni Multiflori Radix ethanol extract (PME) and water extract (PMW).Methods PME and PMW were prepared separately. Male SD rats were randomly assigned into a control group, a positive control (alpha-naphthyl isothiocyanate, ANIT, 100 mg·kg^-1) group, a PMW (15 g·kg^-1) group, and a PME (12 g·kg^-1) group. The rats in the PME and PMW groups were respectively administrated with PME and PMW at the doses equivalent to 60 g·kg^-1 crude drug, which was about 110 times the maximum dose in clinical practice. The ANIT group was intragastrically administrated with ANIT once on the day 40. The serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bile acid (TBA), alkaline phosphatase (ALP), and total bilirubin (TBIL) were determined by kits. Hematoxylin-eosin (HE) staining was carried out for observing the pathological changes of rat liver. Fluorescence quantitative polymerase chain reaction (qRT-PCR) was employed to determine the mRNA levels of bile acid and bilirubin metabolism-related transporters in rat liver tissue after administration, which included organic anion transporting polypeptide 1a1 (OATP1a1), OATP1b2, multidrug resistance-associated protein 2 (MRP2), MRP3, bile salt efflux pump (BSEP), Na⁺-taurocholate co-transporting polypeptide (NTCP), breast cancer resistance protein (BCRP), and P-glycoprotein (P-gp).Results Compared with the control group, PMW lowered the AST level (P<0.05) and elevated the ALT and ALP levels (P<0.05) in the serum; PME lowered the AST level (P<0.05) and elevated the ALT, TBA, and ALP levels (P<0.01) in the serum. There was no significant difference in the level of TBIL between the two groups. The pathological results showed that PME and PMW caused liver injury at different degrees after administration, and more serious lesions were observed in the PME group. The results of qRT-PCR showed that PMW and PME affected the expression of uptake and efflux transporters in the process of bilirubin and bile acid metabolism. Specifically, they significantly down-regulated the mRNA levels of the efflux transporters MRP2 and BCRP and up-regulated those of the uptake transporters OATP1a1 and OATP1b2. Furthermore, PME significantly down-regulated the mRNA levels of BSEP, P-gp, and NTCP; PMW significantly up-regulated the mRNA levels of NTCP, BSEP, and P-gp; PMW and PME had no significant effect on the mRNA level of MRP3.Conclusion After long-term administration, both PME and PMW cause liver injury in rats, which is associated with the effect on bilirubin and bile acid-associated transporters. The degrees of liver injury caused by PME and PMW vary, which may be related to the different effects of the two on specific transporters.