摘要
补骨脂和淫羊藿是临床常用药对,既往研究显示两药可能引起特异质肝损伤,然而其物质基础以及发生机制尚不清楚。该研究基于肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)介导的免疫应激小鼠模型,以补骨脂及淫羊藿中2个主要化学成分补骨脂甲素和朝藿定B为研究对象,通过检测细胞上清液中乳酸脱氢酶(lactate dehydrogenase,LDH)释放量和小鼠血浆中丙氨酸氨基转氨酶(alanine aminotransferase,ALT)、天冬氨酸氨基转氨酶(aspartate transaminase,AST)含量,苏木精-伊红(hematoxylin-eosin,HE)染色观察肝脏病理学变化情况,分别在细胞及动物水平评估补骨脂甲素联合朝藿定B引起肝损伤的程度。借助拟靶向代谢组学技术和多元统计分析方法,考察给药前后小鼠血浆中内源性代谢物的影响,筛选差异代谢标志物,并进行京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)富集通路分析。结果表明,在细胞水平,使用TNF-α刺激2 h后,相较于正常组及单独给药组,补骨脂甲素可以使HepG2细胞中LDH释放量显著增加;补骨脂甲素联合朝藿定B后,LDH释放量在原来基础上进一步显著增加。在动物水平,对于正常小鼠,单独或联合补骨脂甲素均不引起肝损伤;对于免疫应激模型小鼠,单独给药补骨脂甲素或者朝藿定B均可引起肝损伤,补骨脂甲素或者朝霍定B联合给药时肝损伤进一步增强,表现为小鼠血浆中ALT、AST含量显著升高,并伴随肝脏组织大量炎性免疫细胞浸润。基于拟靶向代谢组学技术,筛选得到7个共有差异代谢物,根据受试者操作特性(receiver operating characteristic,ROC)曲线分析结果,其中曲线下面积(area under curve,AUC)大于0.9的包括D-氨基葡萄糖6-磷酸、N1-甲基-2-吡咯-5-甲酰胺、17β-硝基-5a-雄甾烷、葛花苷元-7-O-葡萄糖醛酸苷、N-(1-脱氧-1-果糖基)缬氨酸。因此,这5个差异代谢物可能是潜在的生物标志物。
Chinese patent medicine preparations containing Epimedii Folium and Psoraleae Fructus have been associated with the occurrence of idiosyncratic drug-induced liver injury(IDILI).However,the specific toxic biomarkers and mechanisms underlying these effects remain unclear.This study aimed to comprehensively assess the impact of bavachin and epimedin B,two principal consti-tuents found in Psoraleae Fructus and Epimedii Folium,on an IDILI model induced by tumor necrosis factor-α(TNF-α) treatment,both in vitro and in vivo.To evaluate the extent of liver injury,various parameters were assessed.Lactate dehydrogenase(LDH) release in the cell culture supernatant,as well as the levels of alanine aminotransferase(ALT) and aspartate transaminase(AST) in mouse plasma were measured.Additionally,histological analysis employing hematoxylin-eosin staining was performed to observe liver tissue changes indicative of the severity of liver injury.Furthermore,a pseudo-targeted metabolomics approach was employed,followed by multivariate analysis,to identify differential metabolites.These identified metabolites were subsequently subjected to Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analysis.The results showed that at the cellular level,after 2 hours of TNF-α stimulation,bavachin significantly increased the release of LDH in HepG2 cells compared to the normal group and the group treated alone;after the combination of bavachin and epimedin B,the release of LDH further significantly increased on the original basis.Similarly,although the individual or combination treatments of bavachin and epimedin B did not induce liver injury in normal mice,the combination of both drugs induced marked liver injury in TNF-α treated mice,leading to a significant elevation in plasma AST and ALT levels and substantial infiltration of inflammatory immune cells in the liver tissue.Pseudo-targeted metabolomics analysis identified seven common differential metabolites.Among these,D-glucosamine-6-phosphate,N1-methyl-2-pyridone-5-carboxam
作者
林蒙蒙
李蓥滢
曹波
徐静
肖小河
李国辉
李春雨
LIN Meng-meng;LI Ying-ying;CAO Bo;XU Jing;XIAO Xiao-he;LI Guo-hui;LI Chun-yu(National Cancer Center,National Clinical Research Center for Cancer,Cancer Hospital,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100021,China;the Fifth Medical Center of Chinese PLA General Hospital,Beijing 100039,China)
出处
《中国中药杂志》
CAS
CSCD
北大核心
2024年第2期443-452,共10页
China Journal of Chinese Materia Medica
基金
国家自然科学基金项目(82174071)
中国医学科学院医学与健康科技创新工程项目(2021-I2M-1-014)。
关键词
中成药
补骨脂甲素
朝藿定B
特异质肝损伤
肿瘤坏死因子-Α
拟靶向代谢组学
Chinese patent medicine
bavachin
epimedin B
idiosyncratic liver injury
tumor necrosis factor-α
pseudo-targeted metabolomics
