摘要
目的探讨盐益智仁提取物药液灌胃对肾阳虚多尿大鼠氨基酸代谢的影响及其机制。方法制备盐益智仁及其提取物,采用气相色谱-质谱(GC-MS)技术鉴定提取物成分。选择雄性SD大鼠18只,随机分为对照组、模型组、干预组,每组6只;模型组、干预组均予醋酸泼尼松溶液15 mg/kg和氢氯噻嗪溶液25 mg/kg灌胃制备肾阳虚多尿模型。模型制备成功后,干预组予盐益智仁提取物药液1.04 g/kg灌胃,模型组、对照组予等量0.6%吐温-80灌胃,连续灌胃1周。观察大鼠日常饮食、饮水、排泄、活动量、精神状态等,记录其24h排尿量。末次灌胃6h,收集尿液,用GC-MS技术分析尿液中氨基酸差异代谢物,用紫外-谷氨酸脱氢酶法检测尿液尿素(UREA);腹主动脉取血,用全自动血细胞分析仪检测血常规,用GC-MS技术分析血液中氨基酸差异代谢物;用全自动生化分析仪检测血清肌酐(CREA)、AST、ALT。用MetaboAnalyst6.0在线网站进行代谢通路分析,筛选与氨基酸代谢密切相关的代谢通路;用Pearson偏最小二乘回归法分析氨基酸差异代谢物与药效指标的相关性。结果在盐益智仁提取物中共鉴定出6种主要化学成分,分别为(-)-α-蒎烯、β-芹子烯、马兜铃烯、巴伦西亚橘烯、α-Copaen-15-ol、圆柚酮。与对照组比较,模型组饮水量多,饮食量少,精神萎靡,活动量少,24 h排尿量多;与模型组比较,干预组饮水量少,饮食量恢复,活动量增加、24 h尿量少、精神状态好转。与对照组比较,模型组尿液UREA水平高(P<0.05),血清AST水平低(P<0.05),血清ALT、CREA水平高(P均<0.05),WBC计数低(P<0.05),尿液和血清中氨基酸代谢物水平低(P均<0.05);与模型组比较,干预组尿液UREA水平低(P<0.05),血清AST水平高(P<0.05),血清ALT、CREA水平低(P均<0.05),WBC计数高(P<0.05),尿液和血清中氨基酸代谢物水平高(P均<0.05)。在三组尿液和血清中共发现17种氨基酸差异代谢物。在17种氨基酸差异代谢物中共筛选出5条代谢通路:甘氨酸、丝氨酸和苏氨酸代谢,丙氨酸、天冬氨酸和谷氨酸代谢,半胱氨酸和蛋氨酸代谢,组氨酸代谢,乙醛酸和二羧酸代谢。17种氨基酸差异代谢物与24 h排尿量和血清ALT、CREA水平呈负相关(P均<0.05),与WBC计数、尿液UREA水平和血清AST水平呈正相关(P均<0.05)。结论盐益智仁能够改善肾阳虚多尿大鼠氨基酸代谢和肝肾功能,并且其尿液和血清中氨基酸差异代谢物与药效指标关系密切;盐益智仁的作用机制可能主要通过甘氨酸、丝氨酸和苏氨酸代谢,丙氨酸、天冬氨酸和谷氨酸代谢,半胱氨酸和蛋氨酸代谢,组氨酸代谢,乙醛酸和二羧酸代谢等5条代谢通路调节氨基酸代谢。
Objective To investigate the effect and mechanism of intragastric administration of salt-processed Alpinia oxyphylla extracts liquid on amino acid metabolism in kidney-yang deficiency polyuria rats.Methods The salt-processed Alpinia oxyphylla extracts were prepared and identified by gas chromatography-mass spectrometry(GC-MS).Eighteen male SD rats were randomly divided into the control,model and intervention groups,with 6 rats in each group.Rats in both the model and intervention groups were given prednisone acetate solution 15 mg/kg and hydrochlorothiazide solution 25 mg/kg by gavage to establish the models of kidney-yang deficiency polyuria.After successful preparation of the models,rats in the intervention group were given 1.04 g/kg salt-processed Alpinia oxyphylla extracts,and rats in the model and control groups were given 0.6%Tween-80 by gavage,continuously for 1 week.The daily diet,drinking water,excretion,activity and mental status of the rats were observed,and 24-h urine output was recorded.Urine was collected 6 h after the last gavage,and the differential metabolites of amino acids in urine were analyzed by GC-MS,and UREA was detected by UV glutamate dehydrogenase method.Blood was collected from the abdominal aorta,blood routine was detected by automatic blood cell analyser,and differential amino acid metabolites in blood were analysed by GC-MS.Serum creatinine(CREA),AST and ALT were determined by automated biochemical analyser.Metabolic pathways were analyzed by MetaboAnalyst 6.0 online website,and metabolic pathways closely related to amino acid metabolism were screened.The correlation between amino acid metabolites and pharmacodynamic indexes was analyzed by Pearson partial least squares regression method.Results Six main chemical components were identified in the extracts of salt-processed Alpinia oxyphylla,which were(-)-α-copaene,β-selinene,aristolochene,valencene,α-Copaen-15-ol and nootkatone.Compared with the control group,rats in the model group drank more water and ate less,with worse spirit,less activity and more urine in 24 h.Compared with the model group,rats in the intervention group drank less water with recovered diet,increased activity,decreased urine volume at 24 h and improved mental state.Compared with the control group,the model group had higher urinary UREA(P<0.05),lower serum AST(P<0.05),higher serum ALT and CREA(P<0.05),lower white blood cells(WBC)count(P<0.05),and lower urinary and serum amino acid metabolite levels(P<0.05).Compared with the model group,the intervention group had lower urinary UREA(P<0.05),higher serum AST(P<0.05),lower serum ALT and CREA(P<0.05),higher WBC count(P<0.05)and higher levels of urinary and serum amino acid metabolites(all P<0.05).Seventeen different amino acid metabolites were found in the urine and serum of the three groups.Five metabolic pathways were identified in the 17 different amino acid metabolites:glycine,serine and threonine metabolism,alanine,aspartate and glutamate metabolism,cysteine and methionine metabolism,histidine metabolism,glyoxylic acid and dicarboxylic acid metabolism.The 17 different amino acid metabolites were negatively correlated with 24 h urine output and serum ALT and CREA levels(all P<0.05),but were positively correlated with WBC count,UREA and serum AST levels(all P<0.05).Conclusions The salt-processed Alpinia oxyphylla can improve amino acid metabolism and liver and kidney function in kidney-yang deficiency polyuria rats,and the difference metabolites of amino acids in urine and serum are closely related to pharmacodynamic indexes.The mechanism of action of salt-processed Alpinia oxyphylla may regulate amino acid metabolism mainly through five metabolic pathways such as glycine,serine and threonine metabolism,alanine,aspartate and glutamate metabolism,cysteine and methionine metabolism,histidine metabolism,glyoxylic acid and dicarboxylic acid metabolism.
作者
韩敏
吴珊珊
何席呈
雷艳
王若男
HAN Min;WU Shanshan;HE Xicheng;LEI Yan;WANG Ruonan(College of Pharmacy,Guizhou University of Traditional Chinese Medicine,Guiyang 550025,China)
出处
《山东医药》
2025年第1期48-52,58,共6页
Shandong Medical Journal
基金
国家自然科学基金资助项目(81903806)。
关键词
多尿
肾阳虚
盐益智仁
氨基酸代谢
气相色谱-质谱技术
大鼠
diuresis
kidney-yang deficiency
salt-processed Alpinia oxyphylla
amino acid metabolism
gas chromatography-mass spectrometry
rats
作者简介
第一作者:韩敏(1995-),女,硕士研究生在读,主要研究方向为中药民族药炮制机制。E-mail:1678021541@qq.com;通信作者:何席呈(1986-),男,硕士研究生,助教,主要研究方向为中药民族药质量。E-mail:hexicheng53@163.com。
