With the increasing life expectancy of women worldwide, more and more women enter the perimenopause and postmenopause period, perimenopause syndrome poses a threat to the overall health of women worldwide, and cardiovascular disease is one of the main threats [10, 11]. Studies have shown that cardiovascular disease mortality in premenopausal women is generally lower than that of men of the same age, but since perimenopause, the risk of cardiovascular disease in women has increased sharply [12, 13], and the mechanism of this transition is still unclear. safe. Hormone replacement therapy is the most effective modern medical treatment for PS, but adverse events associated with diseases such as stroke and breast and ovarian cancer cannot be ignored [3, 11]. Therefore, there is an urgent need to provide new treatment options for perimenopausal coronary heart disease without causing adverse effects.
NZEXHJ is a compound preparation developed by our team for the treatment of PS and consists of seven herbs, including Ligustrum lucidum, Curculigo longa, Herba Epimedii, Morinda officinalis, Angelica sinensis, Anemarrhena asphodeloides, and Phellodendron amurense, and has a protective effect on cardiovascular diseases. We have shown through animal experiments that NZEXHJ can protect myocardial tissue, improve cardiac function, reduce myocardial fibrosis and infiltration of inflammatory cells, and protect mitochondrial microstructure in ovariectomized rats with myocardial infarction, but its specific mechanism and target of action are still unclear. To achieve this goal, we explored the mechanism of action of NZEXHJ in the treatment of PMCHD using network pharmacology in conjunction with a molecular docking technique.
One unanticipated result was that 50 pharmacodynamic components of NZEXHJ were obtained by constructing the network analysis of the active ingredients of the perimenopausal targets of NZEXHJ that could act on 52 PS gene targets, among which NZEXHJ significantly acted on SCN5A, SER1, AR, PGR, and other gene targets. The SCN5A gene is a myocardial cell Na + channel that regulates myocardial cell action potential and is involved in the occurrence and development of cardiomyopathy [6]. Genes such as AR [14] and PGR [15] are involved in hormone regulation in the body, thereby improving heart failure symptoms [16], and reducing mortality [17]. Seven core targets, including CASP3, CXCL8 (IL-8), IL6, MAPK1, TNF, TP53, and VEGFA, were finally derived by constructing a PPI network analysis and were considered effective targets in the treatment of PS.
Through network pharmacological analysis, it was found that drugs such as Angeli-cae sinensis Radix, Morindae officinalis Radix, CurculiginisRhizoma, Epimedii Folium, Ligustrilucidi Fructus, Phellodendrichinense Cortex, and Anemarrhenaerhizoma con-tained β-Sitosterol. Quercetin and luteolin are the main active ingredients in the extracts of LigustriLucidi Fructus, Phellodendrichinense Cortex, Epimedii Folium, and Morindae Officinalis Ra-dix, which contain a large number of phenolics and flavonoids and can protect cardiomyocytes through anti-oxidative stress and anti-apoptosis [18–21].Numerous studies have shown that the active ingredients in NZEXHJ, including β-Sitosterol [22], kaempferol [23], luteolin [24], and quercetin [25], can inhibit inflammatory responses and protect cardiomyocytes via a variety of mechanisms. Intervention with NZEXHJ in ovariectomized rats with myocardial infarction revealed that NZEXHJ could reduce the expression of IL-8 and TNF in the heart tissue of ovariectomized rats (IL-8 was significantly decreased). We discovered through network pharmacology research that IL-8 and TNF, which are closely related to inflammation, could serve as the main targets of NZEXHJ. We therefore postulate that NZEXHJ inhibits inflammation in order to protect cardiac tissue.
Additionally, the PI3K-Akt signaling pathway, which is crucial for the occurrence and progression of coronary heart disease, was significantly enriched in the KEGG pathway enrichment analysis, along with lipids and atherosclerosis [26]. NZEXHJ, however, reduced the expression levels of PI3K and p-AKT in the cardiomyocytes of ovariectomized rats with myocardial infarction in animal studies. β-Sitosterol, the active component of NZEXHJ, inhibits PI3K-Akt signaling pathway [27]. Numerous studies have shown that the activity of the PI3K-AKT signaling pathway is one of the mechanisms involved in myocardial fibrosis [28]. By inhibiting this path-way, the level of inflammatory cytokines is reduced [29], the progression of myocardial fibrosis slows down [30], and the apoptosis of myocardial cells is mitigated [31], thereby protecting myocardial tissue. In contrast, the activation of this pathway is closely associated with myocardial injury and even aggravates myocardial cell injury after acute myocardial infarction [28, 32]. Therefore, we speculated that NZEXHJ protected against cell injury leading to myocardial infarction in ovariectomized rats by regulating the PI3K-Akt signaling pathway.
In this experiment, after intervention of ovariectomized rats with myocardial infarction using NZEXHJ, it was also found that ovariectomized rats had reduced uterine weight and increased body weight, similar to the findings of Janas et al [33], and the occurrence of this phenomenon was related to the fact that estrogen regulates insulin sensitivity through the hypothalamus, regulates insulin secretion, and affects the peripheral deposit of adipose tissue [34]. We demonstrated that the continuity of the femoral bone was interrupted after ovariectomy, and NZEXHJ exerted protective effects in this regard. The active ingredients in NZEXHJ (kanamycin, luteolin, and mistletoe) could regulate autophagy and apoptosis of rat osteocytes [35–37], significantly improve the bone mineral density of patients through the AMPK/SIRT1 signaling pathway [38].
In summary, we identified the main target of NZEXHJ in the treatment of PS using network pharmacology and further explored the effects of NZEXHJ in ovariectomized myocardial-infarction rats in order to illustrate the affected target genes (CXCL8 and TNF) and to demonstrate its influence on the PI3K-AKT signaling pathway to prevent myocardial infarction injury in ovariectomized rats.