[Mechanism of Trichosanthis Pericarpium in treating intermingled phlegm and blood stasis syndrome of coronary heart disease based on transcriptomics and metabolomics].

This study explored the potential mechanism of Trichosanthes Pericarpium(TP) in the treatment of intermingled phlegm and blood stasis syndrome of coronary heart disease(CAD-TY) through transcriptomics and metabolomics. SD rats were randomly divided into a control group, a model group, a positive drug Salvia miltiorrhiza group, a high-dose TP group, and a low-dose TP group. The CAD-TY model was prepared by using three methods: infusion of high fat emulsion, intraperitoneal injection of streptozotocin, and coronary artery ligation. The activity or content of total cholesterol(TC), triglyceride(TG), low density lipoprotein(LDL), high density lipoprotein(HDL), blood glucose(GLU), creatine kinase(CK), creatine kinase isoenzyme(CK-MB), and troponin I(cTnI) in plasma was detected by enzyme-linked immunosorbent assay and biochemical kit. Transcriptomics and metabolomics techniques were used to detect differentially expressed genes and differential metabolites in heart tissue. Enrichment analyses were conducted through Kyoto Encyclopedia of Genes and Genomes(KEGG). The expressions of key genes and proteins were verified by reverse transcription-quantitative polymerase chain reaction(RT-qPCR) and Western blot. The results showed that compared with the control group, the model group exhibited severe glucolipid metabolism disorder and myocardial infarction. The activity or content of TC, TG, LDL, GLU, CK, CK-MB, and cTnI in plasma was significantly increased, while the content of HDL was significantly decreased. TP intervention could reverse these pathological changes in model rats. A total of 375 potential genes of TP for the treatment of CAD-TY were identified by transcriptomic analysis. These genes were mainly enriched in signal pathways such as adenosine 5'-monophosphate-activated protein kinase(AMPK), forkhead box O(FoxO), and insulin resistance. Metabolomic analysis showed that TP could reverse the content of 74 metabolites in CAD-TY rats, mainly including lipid metabolites such as enoylcarnitine, corticosterone, and other endogenous components. The metabolites were enriched in metabolic pathways related to glycolipid metabolism, including steroid hormone biosynthesis, lipolysis, and glycolysis/gluconeogenesis. Based on the analysis of transcriptome and metabolome results, the key targets enriched in the AMPK signaling pathway were verified. The results of RT-qPCR detection showed that compared with those of the control group, the expressions of key genes sestrin 1(SESN1), AMP-activated protein kinase catalytic subunit alpha-2(PRKAA2), peroxisome proliferator-activated receptor gamma coactivator 1 alpha(PPARGC1A), insulin receptor(INSR), FoxO3, and Unc-51 like kinase 1(ULK1) mRNAs were significantly decreased in the model group(P<0.01). Compared with those of the model group, the mRNA expressions of the above genes in the hearts of rats in the TP group were significantly increased(P<0.01). The results of Western blot showed that TP could significantly increase the expressions of SESN1, PRKAA2, and PPARGC1A proteins(P<0.01). The detection results of RT-qPCR and Western blot were consistent with those of the transcriptomic analysis. The above research results indicate that TP can significantly improve the glycolipid metabolism disorder of CAD-TY rats and thereby has a protective effect on the damaged myocardium. TP may exert a cardiac protective effect on CAD-TY by upregulating the expression of key targets such as SESN1, PRKAA2, and PPARGC1A, activating the AMPK signaling pathway, promoting acylcarnitine production and fatty acid β oxidation, improving cardiac lipid metabolism, and alleviating myocardial injury. The study partially reveals the biological connotations of TP in "broadening the chest and resolving masses as well as resolving phlegm and freeing channels".