目的 评价补骨脂挥发油的肝毒性作用并分析其潜在毒性机制。方法 采用肝脏具特异性荧光的Tg(fabp10:EGFP)转基因斑马鱼模型评价补骨脂挥发油对肝脏形态、荧光强度、生化指标及组织病理的影响；利用气相色谱-质谱（GCMS）对补骨脂挥发油化学成分进行系统表征；通过成分占比和分子对接毒性系数加权，运用网络毒理学策略筛选补骨脂挥发油肝毒性的核心靶点，并利用基因本体（GO）及京都基因与基因组百科全书（KEGG）通路富集分析其可能的毒性机制，通过构建“药物-成分-靶点-通路”网络筛选出补骨脂挥发油潜在的肝毒性成分和靶点；利用实时荧光定量PCR(qRT-PCR)方法对相关核心靶点进行验证。结果 补骨脂挥发油可显著降低斑马鱼肝脏荧光面积及强度（P<0.001），显著增加丙氨酸氨基转移酶（ALT）、天冬氨酸氨基转移酶（AST）和丙二醛（MDA）水平（P<0.05），降低谷胱甘肽（GSH）水平（P<0.001），并诱导肝细胞核萎缩、空泡化等病理损伤；GC-MS共鉴定出补骨脂挥发油的55种成分，以萜类化合物为主，其中42种为首次从补骨脂中报道；加权网络毒理学共筛选出细胞色素P450 3A4酶（CYP3A4）、周期蛋白依赖性激酶（CDK1）、热休克蛋白90α型1(HSP90AA1)、低氧诱导因子1A(HIF1A)、磷脂酰肌醇4,5-二磷酸3-激酶催化亚基α(PIK3CA)5个与补骨脂挥发油肝毒性相关的核心靶点，毒性机制与细胞衰老、自噬、HIF-1信号通路等相关，氧化石竹烯、(-)-顺式-桃金娘醇、蓝桉醇等成分为补骨脂挥发油中潜在的肝毒性成分；qRT-PCR结果显示补骨脂挥发油处理后斑马鱼中CDK1、HIF1A基因表达显著增加（P<0.001）,PIK3CA基因表达显著减少（P<0.001）。结论 补骨脂挥发油具有一定的肝毒性，其机制与调控核心靶点、干预相关信号通路及毒性成分介导有关。

Objective To evaluate the hepatotoxicity of Psoralea corylifolia volatile oil and analyze its potential toxicity mechanism. Methods The liver-specific fluorescent Tg(fabp10:EGFP) transgenic zebrafish model was employed to evaluate the effects of P. corylifolia volatile oil on liver morphology, fluorescence intensity, biochemical indicator and histopathology. GC-MS was utilized for systematic characterization of the chemical constituents in P. corylifolia volatile oil. By weighting the toxicity coefficients of components and molecular docking, the core targets of P. corylifolia volatile oil hepatotoxicity were screened using the network toxicology strategy, and the possible toxicity mechanisms were investigated with Gene Ontology(GO) and Kyoto Encyclopaedia of Genes and Genomes(KEGG) pathway enrichment analyses. Based on the “drug-component-target-pathway” network, the potential toxicity components and targets of P. corylifolia volatile oil were screened. qRT-PCR method was used to verify the relevant core targets. Results P. corylifolia volatile oil significantly reduced the area and intensity of liver fluorescence(P < 0.01), increased alanine aminotransferase(ALT), aspartate aminotransferase(AST) and malondialdehyde(MDA) levels(P < 0.05), decreased glutathione(GSH) levels(P < 0.001), and induced pathological damage such as liver cell nuclear atrophy and vacuolation. A total of 55 components of P. corylifolia volatile oil were identified by GC-MS, and terpenoids were the main components, of which 42 were reported from P. corylifolia for the first time. Five core targets, Cytochrome P450 3A4 enzyme(CYP3A4), cyclin-dependent kinase 1(CDK1), heat shock protein 90 alpha type 1(HSP90 AA1), hypoxia-inducible factor 1A(HIF1A), and phosphatidylinositol 4,5-bisphosphate 3-kinase catalytic subunit alpha(PIK3 CA), related to the hepatotoxicity of P. corylifolia volatile oil were screened from weighted network toxicology. The toxic mechanism was related to cellular senescence, autophagy, HIF-1 signaling pathway and so on. Caryophyllene oxide,(-)-cis-myrtanol, globulol and other components were potential hepatotoxic components of P. corylifolia volatile oil. The results of qRT-PCR showed that the expression of CDK1 and HIF1A genes in zebrafish increased significantly(P < 0.001) and the expression of PIK3 CA gene decreased significantly(P < 0.001) after treatment with P. corylifolia volatile oil. Conclusion The volatile oil from P. corylifolia exhibits certain hepatotoxicity, and its mechanism is related to the regulation of core targets, intervention in relevant signaling pathways, and mediation by toxic components.

R285.5

山东省中医药科技项目(Z-2023005T)