甘遂醋炙减毒机制的主要途径及靶点预测

Main ways and target prediction of detoxification mechanism of Euphorbia kansui after stir-frying with vinegar

目的研究甘遂醋炙减毒机制的主要途径及靶点。方法取24只SPF级SD大鼠,随机分为空白组、生甘遂组(850 mg/kg)、醋甘遂组(850 mg/kg),每组8只。空白组灌胃0.5%羧甲基纤维素钠溶液,生甘遂组、醋甘遂组给予相应的供试品药液,连续给药20 d,第20天收集大鼠12 h尿液。采用超高效液相色谱串联四极杆静电场轨道阱高分辨质谱(UPLC-Q-Exactive-MS)技术分析尿液样本。经Compound Discoverer 3.0软件进行数据前处理,经BioCyc、HMDB等数据库进行代谢物结构鉴定,应用主成分分析(PCA)、正交偏最小二乘法-判别分析(OPLS-DA)等多种方法观察不同组别是否呈现各自聚类现象;基于MetaboAnalyst进行通路分析,对甘遂醋炙减毒机制潜在靶点进行预测。结果鉴定出20个有显著性差异的内源性代谢物,其中有N-乙酰基-L-天冬氨酸、3-磷酸羟基丙酮酸等10个目标生物标志物为醋甘遂减毒机制作用靶点,参与的主要代谢通路包括精氨酸生物合成,丙氨酸、天冬氨酸和谷氨酸代谢,半胱氨酸和蛋氨酸代谢及精氨酸和D-鸟氨酸代谢。对通路中所有相关代谢物的生物学意义进行分析推测,甘遂醋炙后可能通过降低N-乙酰基-L-天冬氨酸水平缓解神经毒性,通过升高L-高精氨酸水平对心脑血管系统存在保护作用等。结论甘遂经醋炙后,可对机体的神经系统、心脑血管系统、免疫系统及能量代谢等方面的不利因素有明显改善,其发挥减毒机制的最集中相关的代谢通路为精氨酸生物合成。

OBJECTIVE To study main way and target of Euphorbia kansui after stir-frying with vinegar.METHODS Twenty-four SPF grade SD rats were randomly divided into blank group,E.kansui group(850 mg/kg)and vinegar stir-fried E.kansui group(850 mg/kg),with 8 rats in each group.Blank group was given 0.5%sodium carboxymethyl cellulose solution intragastrically,and E.kansui group and vinegar stir-fried E.kansui group were given relevant test sample for consecutive 20 d.The rats’urine of 12 hours was collected on the 20th day.The urine samples of rats in each group were determined by UPLC-QExactive-MS.The data was pre-processed by Compound Discoverer 3.0 software,and the metabolite structure was identified by BioCyc,HMDB and other databases.Whether different groups presented their own clustering phenomenon was observed by principal component analysis(PCA)and orthogonal partial least squares-discriminant analysis(OPLS-DA),etc.Based on the pathway analysis of MetaboAnalyst,the potential targets of detoxification mechanism of E.kansui after stir-frying with vinegar were predicted.RESULTS Twenty significantly differential endogenous metabolites were identified,of which 10 target metabolites,such as N-acetyl-L-aspartate and 3-phosphonooxypyruvic acid,were targets of detoxification mechanism of E.kansui after stirfrying with vinegar.The main metabolic pathways included arginine biosynthesis,alanine,aspartic acid and glutamic acid metabolism,cysteine and methionine metabolism,and arginine and D-ornithine metabolism.The biological significance of all related metabolites in the pathways was analyzed and speculated;after stir-frying with vinegar,E.kansui may alleviate neurotoxicity by reducing the level of N-acetyl-L-aspartic acid;E.kansui had a protective effect on cardio-cerebrovascular system by increasing the level of L-high arginine.CONCLUSIONS After stir-frying with vinegar,E.kansui can significantly improve the adverse factors in terms of nervous system,cardio-cerebrovascular system,immune system and energy metabolism.The most concentrated metabolic pathway related to its detoxification mechanism is arginine biosynthesis.