基于血液代谢组学的升陷汤抗慢性心力衰竭的作用机制研究

Mechanism of Shengxian decoction in treatment of heart failure based on serum metabolomics

目的采用代谢组学技术研究升陷汤治疗慢性心力衰竭的作用机制.方法将大鼠按随机数字表法分为空白组、模型组和给药组,每组10只.模型组和给药组大鼠腹腔注射阿霉素制备心力衰竭大鼠模型.造模后给药组灌胃升陷汤液3.83 g/kg,空白组和模型组灌胃等体积蒸馏水,1次/d,连续灌胃4周.检测大鼠血清肌酸激酶(creatine kinase,CK)、AST、LDH、MDA水平;采用高分辨串联质谱采集各组血清代谢物图谱,利用主成分分析和偏最小二乘判别分析对数据进行统计,通过通路富集分析获取代谢通路,聚焦关键代谢酶和代谢途径.结果与模型组比较,给药组大鼠血清CK[(1 015.44±201.49)U/L比(1 301.89±311.54)U/L]、AST[(210.59±80.34)U/L比(421.56±120.32)U/L]、LDH[(1 211.64±416.61)U/L比(601.58±311.74)U/L]水平降低(P<0.05),MDA[(209.21±151.15)nmol/ml比(1 251.15±110.64)nmol/ml]水平升高(P<0.05);血液代谢轮廓显示,空白组大鼠与模型组大鼠代谢分布显著分离,且给药组处于二者之间.降维后通过偏最小二乘判别分析得到心力衰竭大鼠血液生物标志物,包括柠檬酸、琥珀酸、苹果酸、花生四烯酸、犬尿氨酸、丝氨酸、鞘氨醇、Ceramide(d18:0/14:0)、Sphingomyelin(d18:1/22:0)、Sphingomyelin(d18:0/16:1)、Sphingomyelin[d18:0/18:1(11Z)],MetPA分析显示,心力衰竭大鼠代谢异常主要涉及花生四烯酸代谢、甘氨酸、丝氨酸和苏氨酸代谢、鞘脂代谢、柠檬酸代谢、氨酰基-tRNA生物合成.结论升陷汤对心力衰竭大鼠心肌有一定的保护作用.调节花生四烯酸代谢、甘氨酸、丝氨酸和苏氨酸代谢、鞘脂代谢、柠檬酸代谢、氨酰基-tRNA生物合成等可能是其治疗心力衰竭的关键机制.

Objective To explore the mechanism of Shengxian decoction in the treatment of heart failure by using metabolomic technology. Methods Rats were randomly divided into the control group, model group and administration group according to body weight, with 10 rats in each group. Rats in model group and administration group were induced by intraperitoneal injection of adriamycin to duplicate rat heart failure model. The rats in the treatment group were given Shengxian decoction 3.83 g/kg, while those in the control group and model group were given distilled water of equal volume once a day for 4 weeks. The levels of CK, AST, LDH and MDA in serum of rats were detected by ultraviolet spectrophotometer, and the metabolite profiles were collected by high resolution tandem mass spectrometry. The data were analyzed by principal component analysis and partial least squares discriminant analysis. Metabolic pathways were obtained by pathway enrichment analysis, focusing on key metabolic enzymes and metabolic pathways. Results Compared with the model group, the serum CK (1 015.44 ± 201.49 U/L vs. 1 301.89 ± 311.54 U/L), AST (210.59 ± 80.34 U/L vs. 421.56 ± 120.32 U/L), LDH (1 211.64 ± 416.61 U/L vs. 601.58 ± 311.74 U/L) in the administration group significantly decreased (P<0.05), and MDA (209.21 ± 151.15 nmol/ml vs. 1 251.15 ± 110.64 nmol/ml) levels significantly increased (P<0.05). The metabolic distribution of rats in the control group was significantly separated from that in the model group, and the administration group was between the two groups. After dimension reduction, blood biomarkers were obtained by partial least squares discriminant analysis, including citric acid, succinic acid, malic acid, arachidonic acid, canine uric acid, serine, sphingosine, Cer (d18:0/14:0), SM (d18:1/22:0), SM [d18:0/18:1 (11Z)], SM (d18:0/16:1). Metabo Analyst 4.0 analysis showed abnormal metabolism in heart failure rats, which mainly involved arachidonic acid metabolism, glycine, serine and threonine metabolism, sphingolipid metabolism, citric acid metabolism and aminoacyl-tRNA biosynthesis. Conclusions The Shengxian decoction has a good therapeutic effect on heart failure rats. Regulation of arachidonic acid metabolism, glycine, serine and threonine metabolism, sphingolipid metabolism, citric acid metabolism and aminoacyl-tRNA biosynthesis may be the key mechanisms for its treatment of heart failure.