基于转录组学的补骨脂酚致小鼠肝损伤种属差异  

Species Differences of Bakuchiol-induced Liver Injury in Mice Based on Transcriptomics

作　　者：康倩君 涂灿 郭兆娟 李品 蒋冰倩 王停 张晶璇 KANG Qian-jun;TU Can;GUO Zhao-juan;LI Pin;JIANG Bing-qian;WANG Ting;ZHANG Jing-xuan(Beijing Institute of Traditional Chinese Medicine(TCM),National Medical Products Administration Key Laboratory for Research and Evaluation of TCM,Beijing University of Chinese Medicine,Beijing 100029,China)

机构地区：[1]北京中医药大学北京中医药研究院,国家药品监督管理局中医药研究与评价重点实验室,北京100029

出　　处：《中国实验方剂学杂志》2022年第5期77-85,共9页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金面上项目(81773992);中央高校基本科研业务费重点攻关项目(2020-JYB-ZDGG-109)。

摘　　要：目的:探究补骨脂酚在Institute of Cancer Research(ICR)小鼠和昆明(KM)小鼠不同种属之间的肝毒性应答差异。方法:通过急性毒性和亚急性毒性动物实验证实补骨脂酚致小鼠肝毒性的客观表现,运用转录组学比较正常ICR小鼠和KM小鼠2种不同种属小鼠间的差异基因富集通路,在此基础上采用实时荧光定量聚合酶链式反应(Real-time PCR)验证相关通路上关键基因的mRNA表达证实补骨脂酚致肝损伤的种属差异。结果:补骨脂酚亚急性毒性实验,与正常组比较,ICR小鼠血清碱性磷酸酶(ALP),5′-核苷酸酶(5′-NT)含量均有不同程度升高,差异无统计学意义,KM小鼠无明显变化;病理结果显示ICR小鼠以肝细胞肥大为主要病理特征,KM小鼠主要表现为肝细胞脂肪变性。补骨脂酚急性毒性实验,给药3 d,KM小鼠出现竖毛,精神萎靡,濒死,ICR小鼠无明显毒性表现。与正常组比较,KM小鼠(400 mg·kg^(-1))血清丙氨酸氨基转移酶(ALT),天门冬氨酸氨基转移酶(AST)显著升高(P<0.01),肝脏总活性氧(SOD)酶活力显著降低(P<0.01);ICR小鼠(400 mg·kg^(-1))血清5′-NT,胆碱酯酶(CHE)含量显著升高(P<0.01)。ICR小鼠肝脑比增加20.34%,KM小鼠增加29.14%(P<0.01)。ICR小鼠肝脏病理主要表现为肝细胞肥大,KM小鼠以局灶性炎症为主,伴肝细胞肥大、肝细胞脂肪变性。转录组学京都基因和基因组数据库(KEGG)通路和Reactome通路富集分析,发现ICR小鼠和KM小鼠不同种属间差异基因表达主要涉及氧化磷酸化、胆汁分泌、胆汁酸及胆汁酸盐合成和代谢通路。与正常组比较,细胞色素P450家族成员7A1(CYP7A1)在各给药组均显著上调(P<0.01),多耐药相关蛋白(MRP2)在KM小鼠各给药组均显著降低(P<0.01),在ICR小鼠各给药组显著升高(P<0.01);胆汁酸盐输出泵(BSEP)在ICR小鼠急性肝损伤(400 mg·kg^(-1))组明显降低(P<0.05);SHP1在KM小鼠急性肝损伤(400 mg·kg^(-1))组高表达,法尼醇X受体(FXR)在ICR小Objective: To explore the differences in response to bakuchiol-induced hepatotoxicity between Institute of Cancer Research(ICR) mice and Kunming(KM) mice. Method: The objective manifestations of bakuchiol-induced hepatotoxicity in mice were confirmed by acute and subacute toxicity animal experiments,and enrichment pathways of differential genes between normal ICR mice and KM mice were compared by transcriptomics. The real-time quantitative polymerase chain reaction(real-time qPCR)assay was used to verify the mRNA expression of key genes in the related pathways to confirm the species differences of bakuchiol-induced liver injury. Result: In the subacute toxicity experiment,compared with the normal mice,the ICR mice showed increased serum content of alkaline phosphatase(ALP),and 5 ′-nucleotidase(5 ′-NT),without significant difference,and no manifest change was observed in KM mice. Pathological results showed that hepatocyte hypertrophy was the main pathological feature in ICR mice and hepatocyte steatosis in KM mice.In the acute toxicity experiment,KM mice showed erect hair,mental malaise,and near-death 3 days after administration. The levels of serum alanine aminotransferase(ALT)and aspartate aminotransferase(AST)in KM mice(400 mg·kg^(-1))significantly increased(P<0.01),and the activity of total reactive oxygen species(SOD)in liver significantly decreased(P<0.01)compared with those in normal mice,while the serum content of5 ′-NT and cholinesterase(CHE)in ICR mice(400 mg·kg^(-1))were significantly elevated(P<0.01). The liver/brain ratio in ICR mice increased by 20.34% and that in KM mice increased by 29.14%(P<0.01). The main pathological manifestation of the liver in ICR mice was hepatocyte hypertrophy,while those in KM mice were focal inflammation, hepatocyte hypertrophy, and hepatocyte steatosis. Kyoto Encyclopedia of Genes and Genomes(KEGG)and Reactome pathway enrichment analyses showed that the differential gene expression between ICR mice and KM mice was mainly involved in oxidative phosphorylation,bil

关 键 词：中药 补骨脂酚 肝损伤 种属差异 转录组学 

分 类 号：R22[医药卫生—中医基础理论] R242[医药卫生—中医学]