摘要
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)组、PMW组(15 g·kg)、PME组(12 g·kg)。PME、PMW组大鼠给药剂量相当于60 g·kg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(OATP1a1)、OATP1b2、多药耐药相关蛋白2(MRP2)、MRP3、胆盐外排泵(BSEP)、Na~+-牛磺胆酸共转运多肽(NTCP)、乳腺癌耐药蛋白(BCRP)、P-糖蛋白(P-gp)mRNA表达水平。结果:与对照组比较,PMW组大鼠血清中AST水平显著降低(P<0.05),ALT和ALP水平显著升高(P<0.05);PME组大鼠血清中AST水平显著降低(P<0.05),ALT、TBA和ALP水平显著升高(P<0.01);PMW组于PME组TBIL水平差异无统计学意义。病理结果显示,PME、PMW给药后均可引起不同程度的肝损伤,并且PME组表现更为严重。qRT-PCR结果表明,PMW和PME可同时影响胆红素和胆汁酸代谢过程中摄取和外排转运体的表达,均可显著下调外排转运体MRP2、BCRP mRNA表达水平,上调摄取转运体OATP1a1、OATP1b2 mRNA表达水平;PME可显著下调BSEP、P-gp、NTCP mRNA表达水平;PMW可显著上调NTCP、BSEP、P-gp mRNA水平;两者对MRP3 mRNA表达水平影响差异无统计学意义。结论:经过长期诱导后,PME、PMW均可引起大鼠肝损伤,其作用机制与影响胆红素和胆汁酸相关转运体相关,PME和PMW引发肝损伤程度存在差异,可能与两者对具体转运蛋白的作用不同相关。
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)group, a PMW (15 g·kg) group, and a PME (12 g·kg) group. The rats in the PME and PMW groups were respectively administrated with PME and PMW at the doses equivalent to 60 g·kgcrude 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 upregulated 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.
作者
汪祺
杨建波
王莹
李妍怡
文海若
马双成
WANG Qi;YANG Jian-boa;WANG Ying;LI Yan-yi;WEN Hai-ruo;MA Shuang-cheng(National Institutes for Food and Drug Control,Beijing 100050,China;Beijing University of Chinese Medicine,Beijing 100029,China)
出处
《中国现代中药》
CAS
2022年第9期1720-1726,共7页
Modern Chinese Medicine
基金
国家自然科学基金项目(81973476)。
关键词
何首乌醇提物
何首乌水提物
肝损伤
转运体
胆汁酸
胆红素
Polygoni Multiflori Radix ethanol extract
Polygoni Multiflori Radix water extract
liver injury
transporter
bile acid
bilirubin
