摘要
目的:“异病同治”是中医经典治病法则之一,该研究旨在以“异病同治”理论为切入点,采用气相色谱-质谱联用技术(GC-MS)从血清代谢稳态角度阐释茱萸丸对肝内胆汁淤积(IC)和溃疡性结肠炎(UC)“异病同治”的效应机制。方法:分别以低剂量茱萸丸(0.6 g·kg^(-1))和高剂量茱萸丸(1.2 g·kg^(-1))水煎液灌胃给药干预α-萘基异硫氰酸酯(ANIT)诱导的IC大鼠模型和2,4,6-三硝基苯磺酸(TNBS)诱导的UC大鼠模型。IC实验部分:将24只SD大鼠随机分为4组,分别为正常组、ANIT模型组、茱萸丸低剂量组、茱萸丸高剂量组;UC实验部分:将24只SD大鼠随机分为4组,分别为正常组、TNBS模型组、茱萸丸低剂量组、茱萸丸高剂量组。首先,通过大鼠血清生化指标丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)、γ-谷氨酰转肽酶(γ-GT)、总胆汁酸(TBA)及结肠损伤评分、结肠重量指数、疾病活动指数和组织病理变化分别评价两种疾病模型及茱萸丸对2种疾病的干预效应。其次,利用气相色谱-质谱(GC-MS)仪对大鼠血清进行测序分析,筛选茱萸丸干预两种疾病的共同核心通路,并通过实时荧光定量聚合酶链式反应(Real-time PCR)检测通路中核心基因的表达情况,最终阐释茱萸丸对上述2种疾病模型“异病同治”的生物学机制。结果:IC实验部分,与正常组比较,ANIT模型组大鼠ALT、AST、γ-GT和TBA水平显著升高(P<0.01);与ANIT模型组比较,茱萸丸低剂量组ALT和TBA水平显著降低(P<0.01),茱萸丸高剂量组ALT和TBA水平显著降低(P<0.01),AST和γ-GT水平显著降低(P<0.01)。UC实验部分,与正常组比较,TNBS模型组大鼠结肠损伤评分、结肠重量指数、疾病活动指数显著升高(P<0.01);与TNBS模型组比较,茱萸丸低剂量组结肠重量指数显著降低(P<0.01),疾病活动指数明显降低(P<0.05),茱萸丸高剂量组结肠损伤评分、结肠重量指数、疾病活动指数显著降低(P<0.01)。GC-MS代谢组学分析结合Real-time PCR结果显示,茱萸丸对上述2种疾病模型中精氨酸生物合成紊乱具有相似的反向调控作用。结论:茱萸丸对IC和UC均具有确切的治疗效应,对精氨酸生物合成通路的调控是茱萸丸对上述2种疾病“异病同治”的核心机制。
Objective:The theory of homotherapy for heteropathy is one of the classical rules in traditional Chinese medicine.Taking this theory as a breakthrough point,this study employed gas chromatography-mass spectrometry(GC-MS)to elucidate the mechanism underlying the therapeutic effects of Zhuyuwan on both intrahepatic cholestasis(IC)and ulcerative colitis(UC)from the viewpoint of serum metabolic homeostasis.Methods:The rat models ofα-naphthylisothiocyanate(ANIT)-induced cholestasis and 2,4,6-trinitro-benzenesulfonic acid(TNBS)-induced UC were treated with low(0.6 g·kg^(-1))and high(1.2 g·kg^(-1))doses of Zhuyuwan by gavage.In the experiment regarding IC,24 Sprague-Dawley(SD)rats were randomly assigned into four groups:normal,ANIT model,low-dose Zhuyuwan,and high-dose Zhuyuwan.In the experiment regarding UC,24 SD rats were randomly allocated into four groups:normal,TNBS model,low-dose Zhuyuwan,and high-dose Zhuyuwan.Firstly,the two disease models and the intervention effects of Zhuyuwan on the two diseases were evaluated based on serum levels of biochemical indicators[alanine aminotransferase(ALT),aspartate transaminase(AST),γ-glutamyltranspeptidase(γ-GT),and total bile acid(TBA)],colon damage score,colon weight index,disease activity index,and histopathological changes in rats.Secondly,the rat serum samples were analyzed by gas chromatography-mass spectrometry(GC-MS)to screen the common core pathways of the two disease models,and the expression of core genes in the pathways was determined by Real-time PCR,on the basis of which the biological mechanism of the treatment of the two disease models by Zhuyuwan was ultimately elucidated.Results:The results of the experiment regarding IC showed that the ANIT model group had higher ALT,AST,γ-GT,and TBA levels than the normal group(P<0.01).Compared with the ANIT model group,the low-dose Zhuyuwan group showed declined ALT and TBA levels(P<0.01)and the high-dose Zhuyuwan group showed lowered ALT,TBA,AST,andγ-GT levels(P<0.01).The results of the experiment regarding UC showed that compared with the normal group,the TNBS model group presented increases in the colonic damage score,colon weight index,and disease activity index(P<0.01).Compared with the TNBS model group,the low-dose Zhuyuwan group showcased declines in colon weight index(P<0.01)and disease activity index(P<0.05),and the high-dose Zhuyuwan group showed reductions in the colon damage score,colon weight index,and disease activity index(P<0.01).GC-MS metabolomics analysis combined with qRT-PCR demonstrated that Zhuyuwan had a similar inverse regulatory effect on arginine metabolism disruption in the above two disease models.Conclusion:Zhuyuwan exhibited definite therapeutic effects on both IC and UC,and the regulation of arginine biosynthesis pathway is the core mechanism for the treatment of both diseases by Zhuyuwan.
作者
韩军
文跃强
许宗颖
罗丹
周黎
李雪怡
代雨凡
杨乐乐
沈涛
于瀚
HAN Jun;WEN Yueqiang;XU Zongying;LUO Dan;ZHOU Li;LI Xueyi;DAI Yufan;YANG Lele;SHEN Tao;YU Han(School of Basic Medical Sciences,Chengdu University of Traditional Chinese Medicine(TCM),Chengdu 611137,China;School of Basic Medical Sciences,Guangzhou University of Chinese Medicine,Guangzhou 510006,China;Deng Zhongjia Inheritance Studio,Chengdu University of TCM,Chengdu 611137,China)
出处
《中国实验方剂学杂志》
北大核心
2025年第13期46-53,共8页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(82374331)
四川省中医药管理局中医药科研专项(2024MS161)
四川省科技创新苗子工程培育项目(MZGC20240077)。
关键词
茱萸丸
异病同治
肝内胆汁淤积症
溃疡性结肠炎
代谢组学
精氨酸生物合成
Zhuyuwan
homotherapy for heteropathy
intrahepatic cholestasis
ulcerative colitis
metabonomics
arginine biosynthesis
