基于非靶向代谢组学与转录组学探究氨基酸缓解甘草酸苷致大鼠假性醛固酮增多症的分子机制

Exploring molecular mechanism of amino acids alleviating glycyrrhizin induced by pseudo-hyperaldosteronism in rats based on non-target metabolomics and transcriptomics

目的研究复方甘草酸苷片中氨基酸缓解甘草酸苷致假性醛固酮增多症的机制。方法SD大鼠灌胃给予过量甘草酸苷建立假性醛固酮增多症模型,将SD大鼠分为空白对照组(等体积生理盐水)、甘草酸苷组(甘草酸单铵盐,300 mg/kg)、甘草酸苷与氨基酸复方组(300 mg/kg,含甘草酸单铵盐﹕甘氨酸﹕DL-甲硫氨酸=1.25∶1∶1)连续灌胃14 d,收集大鼠肾脏、肝脏,HE染色法检测大鼠肾脏组织、肝脏组织损伤情况;采用超高效液相色谱-串联飞行时间质谱联用仪(UHPLC-Q-TOF MS)对大鼠肝脏进行非靶向代谢组学分析,筛选潜在靶点;通过转录组学数据分析与醛固酮增多症相关的醛固酮腺瘤患者差异基因功能,关联推测氨基酸改善醛固酮增多的可能机制。结果与空白对照组相比,甘草酸苷组大鼠肾小管扩张、近曲小管上皮水肿;肝细胞空泡样变性、甘草酸苷与氨基酸复方组大鼠肾脏组织、肝脏组织损伤情况有所改善,仅部分大鼠肾小管扩张、肾近曲小管上皮水肿;肝细胞空泡样变性。代谢组学分析,与甘草酸苷组相比,甘草酸苷与氨基酸复方组筛选出9个潜在靶点;转录组学数据分析氨基酸可能通过脂代谢过程的调节等过程改善醛固酮增多。结论氨基酸可通过作用靶点LDLR调节脂代谢通路改变肉豆蔻酸等代谢物的表达缓解假性醛固酮增多症。

Objective The mechanism of pseudo-hyperaldosteronism caused by glycyrrhizin was investigated by amino acids in compound glycyrrhizin tablets.Methods SD rats were given glycyrrhizin by gavage to establish a model of pseudo-hyperaldosteronism,and SD rats were divided into blank control group(equal volume of saline),glycyrrhizin group(Monoammonium Glycyrrhizinate,300 mg/kg),and glycyrrhizin and amino acid compound group(300 mg/kg,Monoammonium Glycyrrhizinate:Glycine:DL-methionine=1.25∶1∶1)were gavaged continuously for 14 d.The kidneys and livers of the rats were collected with HE stained were used to detect kidney and liver tissue damage;the liver of the rats were analyzed metabolomically by ultra-high performance liquid chromatography-tandem time-of-flight mass spectrometry(UHPLC-Q-TOF MS)to screen potential targets;transcriptomic data were used to analyze the potential targets associated with aldosteronism-related differential gene function in patients with aldosteronism adenoma,and correlation speculation of possible mechanisms of amino acid amelioration of aldosteronism.Results Compared with blank control group,renal tubular dilatation and proximal tubular epithelial edema were observed in glycyrrhizin group;hepatocyte vacuole-like degeneration,renal tissue and liver tissue damage were improved in glycyrrhizin and amino acid combination group of the rats,and part of the rats had renal tubular dilatation,renal proximal tubular epithelial edema and hepatocyte vacuole-like degeneration.In the metabolomic analysis,9 potential targets were screened in glycyrrhizin and amino acid combination group compared with glycyrrhizin group;the transcriptomic data analyzed that amino acids may improve aldosterone increase through the processes such as regulation of lipid metabolic processes.Conclusion Amino acids may alleviate pseudo hyperaldosteronism by modulating lipid metabolic pathways through acting on the target LDLR to alter the expression of metabolites such as myristic acid.