摘要
目的结合代谢组学技术观察大黄-附子药对治疗阳虚便秘模型大鼠的效果,探讨其机制和关键代谢通路。方法将40只SD大鼠随机分为空白对照组、模型对照组、大黄单煎组、附子单煎组、大黄-附子合煎组,每组8只。除空白对照组外,其余各组建立阳虚便秘大鼠模型。空白对照组和模型对照组予煮沸后的超纯水灌胃,其余各组予相应药物煎液灌胃(药物浓度均为1 g生药/mL),各组灌胃剂量均为1 mL/200 g体质量,共计给药4 d。观察各组大鼠的体温、小肠推进率和血清胃动素(MTL)水平;苏木精-伊红染色观察大鼠结肠组织病理改变;基于液相色谱-质谱联用技术对大鼠代谢产物进行模式识别与分析,鉴定差异代谢物并分析其富集的代谢通路。结果与模型对照组比较,大黄-附子合煎组大鼠在给药4 d后,肛温[(38.17±0.60)℃比(36.07±0.25)℃,P<0.05]、额温[(35.77±0.06)℃比(35.27±0.15)℃,P<0.05]、小肠推进率[(73.58±2.47)%比(43.56±2.79)%,P<0.01]、血清MTL[(350.96±15.91)pg/mL比(280.97±25.13)pg/mL,P<0.05]明显升高,差异有统计学意义。各组大鼠结肠病理显示,大黄-附子合煎组与空白对照组的肠壁厚度最为接近。代谢组学分析结果:大黄-附子合煎组与模型对照组大鼠代谢谱明显分离,最终筛选出尿嘧啶、牛磺酸、磷酸胆碱、三甲胺、2-羟基丁酸、丙酸、乙醇、色氨酸、肌氨酸、异丁酸盐、甘氨酸11种内源性差异代谢物,主要富集于甘氨酸、丝氨酸和苏氨酸代谢通路,乙醛酸和二羧酸代谢通路,苯丙氨酸、酪氨酸和色氨酸的生物合成代谢通路。结论大黄-附子药对对阳虚便秘大鼠模型有确切疗效,可能通过上调血清MTL和影响甘氨酸、丝氨酸和苏氨酸代谢通路,乙醛酸和二羧酸代谢通路,苯丙氨酸、酪氨酸和色氨酸的生物合成代谢通路发挥治疗作用。
Objective This study aimed to observe the therapeutic effect of rhubarb-aconite drug pair on yang deficiency constipation in model rats using metabolomics technology and to explore its underlying mechanisms and key metabolic pathways.Methods Forty SD rats were randomly divided into the blank control group,the model control group,the rhubarb single decoction group,the aconite single decoction group,and the rhubarb-aconite combined decoction group,with 8 rats in each group.Except for the blank control group,all other groups were induced to establish rat models of yang deficiency constipation.The blank control and model control groups received boiled ultrapure water by gavage,while the other groups received the corresponding decoctions by gavage(each decoction concentration was 1 g crude drug/mL).The gavage dosage for all groups was 1 mL/200 g of body weight,administered for 4 days.The body temperature,small intestine propulsion rate,and serum motilin(MTL)levels of the rats were measured.Hematoxylin-eosin staining was used to observe pathological changes in the colon tissue.Metabolic profiling and analysis were performed using liquid chromatography-mass spectrometry technology to identify differential metabolites and analyze their enriched metabolic pathways.Results Compared with the model control group,after 4 days of treatment,the rhubarb-aconite combined decoction group showed significant increases in rectal temperature[(38.17±0.60)℃vs.(36.07±0.25)℃,P<0.05],forehead temperature[(35.77±0.06)℃vs.(35.27±0.15)℃,P<0.05],small intestinal propulsion rate[(73.58±2.47)%vs.(43.56±2.79)%,P<0.01],and serum MTL[(350.96±15.91)pg/mL vs.(280.97±25.13)pg/mL,P<0.05].Pathological examination of the colon showed that the intestinal wall thickness in the rhubarb-aconite combined decoction group was closest to that of the blank control group.Metabolomics analysis revealed distinct metabolic profiles between the rhubarb-aconite combined decoction group and the blank control group.Finally,11 endogenous differential metabolites were identified,including uracil,taurine,choline phosphate,trimethylamine,2-hydroxybutyric acid,propionic acid,ethanol,tryptophan,creatine,isobutyrate,and glycine.These metabolites were mainly enriched in the glycine,serine,and threonine metabolism pathways,glyoxylate and dicarboxylate metabolism pathways,and the biosynthesis pathways of phenylalanine,tyrosine,and tryptophan.Conclusion The rhubarb-aconite drug pair has a definite therapeutic effect yang deficiency constipation in a rat model,possibly by upregulating serum MTL and affecting the glycine,serine,and threonine metabolism pathways,glyoxylate and dicarboxylate metabolism pathways,and the biosynthesis pathways of phenylalanine,tyrosine,and tryptophan.
作者
张悦
程苇钇
何莉
任玮晔
朴寄纲
包丹丹
ZHANG Yue;CHENG Weiyi;HE Li;REN Weiye;PIAO Jigang;BAO Dandan(Science and Education Department,Wenzhou Hospital of Integrated Traditional Chinese and Western Medicine,Wenzhou,Zhejiang 325000,China;School of Pharmacy,Zhejiang Chinese Medical University,Hangzhou,Zhejiang 310053,China;Department of Dermatology and Plastic Surgery,the First Affiliated Hospital of Zhejiang Chinese Medical University,Hangzhou,Zhejiang 310003,China)
出处
《浙江中西医结合杂志》
2024年第10期897-904,共8页
Zhejiang Journal of Integrated Traditional Chinese and Western Medicine
基金
浙江省中医药科技计划项目(No.2021ZQ036)。
关键词
大鼠
阳虚便秘
大黄-附子药对
配伍机制
代谢组学
Rat
Yang deficiency constipation
Rhubarb-aconite drug pair
Compatibility mechanism
Metabonomics
