基于P450酶的雷公藤多苷致大鼠肝损伤及其与复方甘草酸苷配伍减毒研究

doi:10.19803/j.1672-8629.20220356

摘要/Abstract

摘要： 目的明确雷公藤多苷（tripterygium glycosides,TG）肝毒性的剂量-时间-效应关系,基于细胞色素P450酶的调控研究TG致大鼠肝损伤的毒性机制,探讨复方甘草酸苷（compound glycyrrhizin,CG）配伍TG的减毒机制,为TG临床安全应用提供理论依据。方法雄性SD大鼠30只,按体重随机分为对照组、TG 189.0 mg·kg^-1组、TG 472.5 mg·kg^-1组、TG 189.0 mg·kg^-1和TG 472.5 mg·kg^-1剂量分别与CG 20.25 mg·kg^-1剂量联合用药组,共5组,每组6只。每日给药1次,连续3周。分别于给药后1、2、3周,测定血清中总蛋白（TP）、白蛋白（ALB）、谷草转氨酶（AST）、谷丙转氨酶(ALT)含量;末次给药结束,计算肝系数,行组织病理学检查,对CYP2B1和CYP3A1基因的转录水平进行检测。结果给药1周后,TG 189.0、472.5 mg·kg^-1组血清中ALB水平均降低（P<0.05）,TG 472.5 mg·kg^-1组ALT升高（P<0.01）;至给药3周,除上述指标的改变外,TG189.0、472.5 mg·kg^-1组进一步引起大鼠血清TP含量的降低（P<0.05）;与CG联合用药3周后,TG472.5 mg·kg^-1剂量联合用药组与同等剂量的TG单独用药组比较,可降低ALT含量（P<0.01）,升高ALB、TP含量（P<0.05）。至给药3周,TG 189.0 mg·kg^-1组的大鼠肝细胞轻微肿胀,汇管区少量淋巴细胞浸润;TG 472.5 mg·kg^-1组的大鼠肝重量和肝系数较对照组增加（P<0.05,P<0.01）,肝被膜下见片状坏死灶伴出血,肝细胞空泡变性、肿胀,汇管区淋巴细胞浸润。与CG联合用药后,大鼠肝脏的组织结构得到修复。189.0、472.5 mg·kg^-1剂量的TG均可导致大鼠肝组织中CYP2B1和CYP3A1 mRNA水平的升高（P<0.05,P<0.01）;TG与CG联合用药后,CYP2B1 mRNA水平下调。结论连续3周给予大鼠189.0、472.5 mg·kg^-1剂量的TG,可对大鼠造成不同程度的肝损伤。其中,TG 472.5 mg·kg^-1组肝损伤更为严重,给药3周较给药1周时肝毒性更显著,具有剂量-时间-效应关系。大鼠肝细胞内细胞色素P450酶CYP2B1和CYP3A1 mRNA过表达为其毒性机制之一;CG可通过下调CYP2B1 mRNA水平,减轻肝毒性。

关键词: 雷公藤多苷, 肝损伤, 药物代谢酶, 复方甘草酸苷, 配伍减毒, 大鼠, P450酶, 大鼠

Abstract: Objective To clarify the dose-time-effect relationship of tripterygium glycosides (TG) hepatotoxicity, investigate the mechanism of TG hepatotoxicity in rats based on the regulation of cytochrome P450 enzymes, and explore the mechanism of toxicity reduction of compound glycyrrhizin (CG) with TG in order to ensure safe clinical applications of TG. Methods Thirty male SD rats were randomly divided into the control group, TG 189.0 mg·kg^-1 group, TG 472.5 mg·kg^-1group, and TG 189.0 and 472.5 mg·kg^-1 respectively combined with CG 20.25 mg· kg^-1 groups according to body weight, with 6 rats in each. The drug was administered once daily for three weeks. The serum levels of total protein(TP)、albumin(ALB), aspartate aminotransferase(AST), and alanine aminotransferase(ALT)were measured at 1, 2, and 3 weeks after dosing, respectively. At the end of the last dose, liver coefficients were calculated and histopathological examinations were performed to detect the transcript levels of CYP2B1 and CYP3A1 genes. Results One week after administration, the serum levels of ALB in the TG 189.0 and 472.5 mg·kg^-1 groups became lower (P<0.05), but ALT in the TG 472.5 mg·kg^-1 group was increased (P<0.01). Three weeks after administration, the level of TP in the TG 189.0, and 472.5 mg·kg^-1 groups continued to decrease. After 3 weeks of coadministration with CG, the ALT level in the TG 472.5 mg·kg^-1 group decreased (P<0.01) while those of ALB and TP (P<0.05) increased compared with the same dose of TG used alone. After three weeks of administration, rats in the TG 189.0 mg·kg^-1 group showed slight swelling of hepatocytes and a small amount of lymphocyte infiltration in the confluent area while those in the TG 472.5 mg·kg^-1 group showed an increase in liver weight and liver coefficient compared with the control group (P<0.05, P<0.01). TG at doses of 189.0 and 472.5 mg·kg^-1 increased CYP2B1 and CYP3A1 mRNA levels in liver tissues of rats (P<0.05, P<0.01). CYP2B1 mRNA levels were down-regulated after the combination of TG and CG. Conclusion TG administered to rats at doses of 189.0 and 472.5 mg·kg^-1 for three consecutive weeks caused different degrees of liver injury. Hepatotoxicity of TG 472.5 mg·kg^-1 was more severe, and was more significant three weeks after administration than one week after administration, with a dose time effect relationship. Overexpression of CYP2B1 and CYP3A1 mRNA in rat hepatocytes is one of the mechanisms of toxicity. CG can reduce hepatotoxicity by downregulating the CYP2B1 mRNA level.

Key words: tripterygium glycosides, liver injury, drug metabolism enzyme, compound glycyrrhizin, compatibility attenuated, rat, P450 enzymes, rat

中图分类号:

R969.3

付小桐, 刘婷, 曹春雨, 张海静, 夏冰, 赵晓昂, 王丽芳, 赵春晖, 杨依霏. 基于P450酶的雷公藤多苷致大鼠肝损伤及其与复方甘草酸苷配伍减毒研究[J]. 中国药物警戒, 2023, 20(1): 85-91.

FU Xiaotong, LIU Ting, CAO Chunyu, ZHANG Haijing, XIA Bing, ZHAO Xiao'ang, WANG Lifang, ZHAO Chunhui, YANG Yifei. Liver injury induced by tripterygium glycosides based on P450 enzymes and toxicity reduction of compound glycyrrhizin[J]. Chinese Journal of Pharmacovigilance, 2023, 20(1): 85-91.

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