We aimed to investigate the protective effects of the active ingredients of Aconite and Glycyrrhiza glabra on rats with CHF. Firstly, we performed ELISA to examine the effects of HA + GA treatment on plasma TCHO and TG levels as well as immunohistochemistry to investigate expression levels of FGF, VEGF and eNOS protein in CHF rats after HA + GA treatment. In addition, LC-MS/MS analysis used to determine the metabolic differences between the HA+GA group and the Model group and to find potential biomarkers and their possible metabolic pathways associated with CHF.
The biochemical tests revealed that TCHO and TG levels in Model were higher than in the HA + GA group, while the difference between the HA + GA and Sham groups was not statistically significant, suggesting that the protective effect of HA + GA on chronic heart failure was associated with a reduction in lipid levels. In hypercholesterolemic patients, plasma FGF2 levels were significantly lower, which was associated with the inhibitory effect of plasma low-density lipoprotein (LDL) [18]. In atherosclerotic lesions, VEGFA expression expressed in smooth muscle cells was significantly increased [19]. It is well known that eNOS has atherosclerotic effects through the synthesis of NO, which has been shown to promote atherosclerosis by oxidizing LDL. The immunohistochemistry results showed that the HA+GA group achieved a protective effect against CHF by up-regulating the expression of FGF2 and VEGFA proteins and attenuating eNOS protein expression.
Sphingolipids are a complex class of lipids consisting of a backbone sphinx base that can be phosphorylated, acylated, glycosylated, bridged to various head groups via phosphodiester bonds, or otherwise modified, resulting in hundreds of sphingolipid subspecies [10]. Sphingolipids have been found to be closely associated with cardiovascular disease; and sphingomyelin levels are significantly increased in atherosclerotic lesions [11], and Poss AM et al., showed that its serum levels have been suggested as a biomarker for cardiovascular disease [12]. Sphingosine is a biologically active signaling molecule among sphingolipid metabolites, and we observed that sphingosine levels were significantly downregulated in the HA + GA group compared to that in the Model group, and we speculated that HA + GA may achieve protective effects in rats with CHF by modulating sphingolipid metabolic pathways and sphingolipid signaling pathways.
Phosphatidylcholine is a common glycerophospholipid that is widely found in animal and plant tissues and egg yolk. The choline component of dietary phosphatidylcholine is metabolized by human intestinal microorganisms to produce trimethylamine-N-oxide (TMAO), which increased the risk of cardiovascular disease [20]. Paapstel K et al. found a negative correlation between serum phosphatidylcholine levels and vascular injury in patients with atherosclerosis [21]. We found that phosphatidylcholine levels were significantly upregulated in the HA+GA group compared to that in the Model group, suggesting that HA + GA achieves vascular protection in CHF rats by promoting the synthesis of phosphatidylcholine. Cardiolipin is a phospholipid specific to mitochondrial membranes and plays an important role in the composition and function of mitochondrial membranes. Impaired energy metabolism is a feature of heart failure, which is associated with iron deficiency and impaired mitochondrial function in patients with heart failure [14]. The important function of cardiolipin for mitochondria lies in its combined action on protein complexes and super complexes [22]. Even for green plants, cardiolipin plays an important role in the mitochondrial development and function [17]. We found that cardiolipin levels were significantly up-regulated in the HA+GA group, indicating that HA+GA achieved a protective effect against CHF in rats by modulating the glycerophospholipid pathway. In addition to regulating sphingolipid metabolic pathway and glycerophospholipid metabolic pathway, it is also related to regulate sphingolipid signaling pathway, apoptosis, arachidonic acid metabolism, linoleic acid metabolism, α-linolenic acid metabolism, and bile secretion pathway.