基于~1H-NMR代谢组学技术分析逍遥丸干预非酒精性脂肪肝肝郁脾虚证大鼠的作用机制

Analysis of Mechanism of Xiaoyaowan on Rats with Nonalcoholic Fatty Liver Disease and Syndrome of Liver Depression and Spleen Deficiency by ~1H-NMR Metabolomics

目的：研究逍遥丸对非酒精性脂肪肝（NAFLD）肝郁脾虚证大鼠尿液中内源性代谢产物的影响,探讨逍遥丸对该病证的作用机制。方法：将SD大鼠随机分为3组,即空白组、模型组、逍遥丸组（3.24 g·kg-1·d-1）,采用高糖高脂饲料结合饥饱失常、慢性束缚应激刺激、夹尾法,共10周复制NAFLD模型,逍遥丸组给予逍遥丸治疗4周。采用酶联免疫吸附法检测模型大鼠血清丙氨酸氨基转移酶、肝组织总胆固醇、甘油三酯、脑组织5-羟色胺和去甲肾上腺素的含量,运用苏木精-伊红染色观察肝组织形态学的变化。采用代谢组学技术对模型大鼠尿液进行核磁数据采集,结合多元统计分析探讨逍遥丸的治疗作用机制。结果：逍遥丸组能使NAFLD肝郁脾虚证大鼠尿液中乙酸,肌酐,甘氨酸,异亮氨酸,α-酮戊二酸,乳酸,亮氨酸,琥珀酸,牛磺酸,二甲胺,α-D-葡萄糖,甘油共12个标志物发生不同程度的回调;主要涉及能量代谢、糖代谢和胰岛素抵抗等代谢途径的调节。结论：逍遥丸可能通过调节脂质代谢、糖代谢和氨基酸代谢紊乱等来发挥治疗NAFLD的作用。

Objective： To study on the effect of Xiaoyaowan on the endogenous metabolites in the urine of rats with nonalcoholic fatty liver disease（ NAFLD） and liver depression and spleen deficiency syndrome,and to explore the therapeutic mechanism of this compound. Method： SD rats were randomly divided into 3 groups,namely blank group,model group and Xiaoyaowan group（ 3. 24 g·kg-1·d-1）. The high-fat and high-fat diet combined with irregular diet, chronic restraint stress stimulation, and tail clipping method were employed to replicate the NAFLD model for 10 weeks. Xiaoyaowan was given for 4 weeks. The levels of serum alanine aminotransferase,total cholesterol and triglycerides in the hepatic tissue,5-hydroxytryptamine and norepinephrine in brain tissue of model rats were detected by enzyme-linked immunosorbent assay. The morphology of liver tissue was observed by hematoxylin-eosin（ HE） staining. The nucleus magnetic data of model rat urine was collected by metabolomics and combined with multivariate statistical analysis to explore the therapeutic mechanism of Xiaoyaowan. Result： Xiaoyaowan could make acetic acid,creatinine,glycine and other nine markers in the urine of rats with NAFLD and syndrome of liver depression and spleen deficiency have varying degrees of callback. It mainly involved the regulation of metabolic pathways,such as energy metabolism,glucose metabolism,insulin resistance,etc. Conclusion： Xiaoyaowan may play a role in the treatment of NAFLD by regulating lipid metabolism,glucose and amino acid metabolism disorders.