Exploring the mechanism of mulberry leaf chrysanthemum in treating hypertension based on network pharmacology and molecular docking
Hypertension is a multi gene genetic disease caused by both environmental and genetic factors, characterized by elevated systemic arterial blood pressure. It can be accompanied by functional or organic damage to organs such as the heart, brain, and kidneys. It is one of the common cardiovascular and cerebrovascular diseases in China and a major risk factor for stroke, coronary heart disease, and heart failure. With the increasing living standards of Chinese residents, hypertension is on the rise. If left untreated, it seriously affects human health and life. Traditional Chinese Medicine classifies hypertension as “headache”, “dizziness”, “wind dizziness” and other categories. The etiology and pathogenesis are often due to the imbalance of qi, blood, yin and yang in the organs, and the coexistence of wind, fire, phlegm, blood stasis, and deficiency. Positive deficiency is the root cause, while evil is the target. In clinical practice, hypertension is characterized by a mixture of deficiency and excess. Traditional Chinese medicine has the characteristics of multi efficacy, increasing efficiency and reducing side effects, multi-target, and forward-looking in treatment. It can protect target organs, reduce toxic side effects, and improve the quality of life of hypertensive patients, playing a unique advantageous role.

Folium Mori (FM) is the dried leaf of Morus alba L. in the family Moraceae, while Flos Chrysanthemi (FC) is the dried inflorescence of Chrysanthem morifolium Ramat. in the family Asteraceae. Both are sweet and cold, with a light body, and enter the lungs and liver meridians. They are commonly used in clinical practice in combination with the upper burner to clear heat and wind, and the lower burner to clear heat and calm the liver, benefit yin and improve vision. In the “Popular Treatise on Cold Damage”, Lingjiao Gouteng Tang is a combination of the two herbs, which can calm the liver and latent yang, clear heat and calm the wind, and help the emperor to clear heat and calm the liver. In clinical practice, the combination of the two can often be used to treat symptoms such as wind heat cold, lung heat cough, initial onset of warm disease, liver yang hyperactivity headache and dizziness, red eye swelling and pain, dark eye dizziness, sores, abscesses, and toxins. Modern doctors often use mulberry leaf chrysanthemum medicine to calm the liver and clear the liver for the treatment of hypertension, which can effectively alleviate symptoms such as headache, dizziness, red eye swelling and pain caused by hypertension. However, the material basis and mechanism of action of mulberry leaf chrysanthemum for lowering blood pressure are not yet clear, and there are few research reports. Therefore, this study adopts network pharmacology and molecular docking methods to analyze the pharmacological substance basis of mulberry leaf chrysanthemum in the treatment of hypertension, explore its potential molecular mechanism of blood pressure reduction, and provide theoretical basis for related basic experimental research and clinical applications.

 

Hypertension is the primary risk factor for stroke and coronary heart disease, and one of the most important factors leading to death worldwide. The etiology and pathogenesis of traditional Chinese medicine are complex, with symptoms such as yin deficiency, yang hyperactivity, wind movement, phlegm dampness and blood stasis obstruction. The disease site is mainly responsible for the liver, kidneys, and spleen. Modern medicine believes that its onset is influenced by multiple links, mechanisms (RAAS, oxidative stress, inflammation, neuroimmune factors, non coding RNA), and multiple factors. Mulberry leaves are the essence of Jixing, which is good for wind, so they are good at searching for wind “(from” Chengconvenient Reading “), while chrysanthemums are” the elite that obtain golden water, which can benefit the two organs of golden water. Moisturizing water can make fire, and benefiting gold can make wood flat “(from” Compendium of Materia Medica “). Mulberry leaves and chrysanthemums are classic medicinal pairs commonly used in clinical practice. Although they are light and flat products, they can benefit the lungs and liver, make kidney water sufficient, nourish liver nature, clear liver heat, and commonly treat dizziness, headache, irritability, facial redness, and red eyes caused by hypertension. However, the specific pharmacological substances and mechanisms of action are still unclear. This study used the network pharmacology research model to screen 41 co acting proteins of mulberry leaf chrysanthemum and hypertension, involving 29 active ingredients. Topological analysis identified 156 core target information and 39 pathways related to hypertension, reflecting the mechanism of the joint action of mulberry leaf chrysanthemum “multi-component, multi-target, and multi pathway” in the treatment of hypertension.
According to Table 2 and Figure 2, the main active ingredients for treating hypertension may include quercetin, kaempferol, β – carotene, arachidonic acid, acacetin, luteolin, etc. Among them, quercetin and kaempferol are shared by mulberry leaves and chrysanthemums. Quercetin can reduce oxidative stress, interfere with the renin angiotensin aldosterone system (RAAS), and improve endothelial dependence and balance of endothelin-1 (ET-1) and NO, thereby improving endothelial function and lowering blood pressure. Kaempferol can endothelium dependent vasodilation, inhibit vascular remodeling caused by angiotensin II, and improve insulin resistance in type II diabetes rats by inhibiting IKK/NF – κ B inflammatory damage IRS-1, and has a lipid-lowering effect on them. β – carotene can reduce the oxidation of low-density lipoprotein (LDL), eliminate free radicals, and prevent vascular damage. Arachidonic acid is the lipid contained in mulberry leaves, which has the effect of preventing atherosclerosis and antithrombotic. Acacia and luteolin are flavonoids found in chrysanthemums. Total flavonoids in chrysanthemums can regulate the NO mediated pathway to control calcium and potassium ion channels, and have protective effects on vasodilation and vasodilation. luteolin also has the ability to scavenge free radicals and protect cells, and synergistically increases antioxidant activity with other antioxidants in the body. In addition, 41 main active ingredient therapeutic targets for hypertension were collected, involving multiple factors such as inflammation, antioxidant, angiogenesis, and vascular remodeling, indicating that mulberry leaf chrysanthemum can exert synergistic antihypertensive effects through multiple components and targets.
Network topology analysis is the multi-level and multi angle mining of relevant information from protein interaction network systems, and the screening of multi-target drug molecules with specific signal nodes, providing reference for predicting the interrelationships between drugs and diseases. This study analyzed protein network interactions and topological parameters of 41 key targets, and obtained a target network with 156 nodes and 3808 edges centered on EGFR, JUN, and PRKCA. On the one hand, key targets were further screened, and on the other hand, the related gene network was expanded, providing evidence for GO functional enrichment, KEGG pathway analysis, and molecular docking validation. According to the topological analysis results in Table 3 and Figure 3, combined with Table 1 and Figures 2 and 6, it can be seen that the key targets include JUN, MMP9, PTGS2, EGFR, CCND1, MMP1, SPP1, PRKCA, ELK1, IGFBP5, KIF5C, SH2D2A, MEP1B, F2, PZP, etc. Studies have shown that the pathogenesis of hypertension is mainly related to the activation of RAAS, hyperactivity of sympathetic nerve, insulin resistance, water and sodium retention, abnormal membrane ion transport and vascular endothelial dysfunction, leading to vasoconstriction, injury, remodeling, and elevated blood pressure. Among them, Jun kinase (JUN) regulates various genes involved in VSMC proliferation, migration, ROS production, and extracellular matrix degradation through transcriptional regulation. Matrix metalloproteinase 9 (MMP9) is involved in the breakdown of extracellular matrix during normal physiological processes and can participate in angiogenesis by releasing vascular endothelial growth factor (VEGF). Meanwhile, the increased activity of matrix metalloproteinase 1 (MMP1) increases the risk of hypertension related microvascular dysfunction. Prostaglandin endoperoxide synthase 2 (PTGS2) is an inducible immediate response gene that can be rapidly upregulated by cytokines, inflammatory mediators, hypoxia, and other stimuli. Its main product, prostaglandin, has biological activities such as promoting cell proliferation and angiogenesis. The epidermal growth factor receptor (EGFR) is widely distributed on the surface of mammalian epithelial cells, fibroblasts, keratinocytes, and other cells, playing an important role in cell growth, proliferation, and differentiation. Functional loss or abnormal activity of key signaling pathway factors can lead to cardiovascular disease and immune deficiency. The cyclin D1 gene (CCND1) has the function of regulating cell cycle, cell growth, and cell differentiation. When cells are damaged, the transcription products of LncRNA upstream of the promoter can bind to TNA binding protein TLS. The protein kinase C – α (PRKCA) gene is a member of the serine threonine specific protein kinase C family, involved in vascular smooth muscle contraction and vascular endothelial growth factor pathway, and involved in calcium ion regulation and vascular smooth muscle contraction. Molecular docking further confirmed the component screening results of network pharmacology, indicating that potential targets PTGS2, MMP9, MMP1 have good binding activity with quercetin and luteolin, and PTGS2 has good binding activity with kaempferol, β – carotene, acacetin, etc. Moreover, the binding conformation is stable, indicating that these may be key components and targets for mulberry leaf chrysanthemum treatment of hypertension.
Perform GO functional and KEGG pathway enrichment analysis on key targets to further explore the mechanism of action of mulberry leaf chrysanthemum in treating hypertension. The results showed that mulberry leaf chrysanthemum may participate in the reaction of lipopolysaccharide, positive regulation of smooth muscle cell proliferation, hypoxia reaction, redox process, regulation of protein catabolism and other biological processes. In extracellular space, endoplasmic reticulum membrane, cell membrane raft and other parts, enzyme binding, serine endopeptidase activity, protein homopolymerization activity, heme binding, protein binding and other molecular reactions occur, and then regulate the IL-17 signaling pathway, fluid shear stress and atherosclerosis signaling pathway, TNF signaling pathway, and microRNA signaling pathway in cancer to play a hypotensive role. The TNF and IL-17 signaling pathways are associated with inflammatory responses. Abnormal regulation of the TNF signaling pathway can lead to damage to endothelial cells, smooth muscle cell proliferation, increased peripheral vascular resistance, and hypertension. The IL-17 signaling pathway can induce the expression of pro-inflammatory cytokines, chemokines, and metalloproteinases in various tissues and cells. Fluid shear stress and atherosclerotic signal pathway are mainly related to hemorheology. Lower blood flow shear stress is easy to lead to the formation of atherosclerosis or plaque rupture, endothelial damage, MMP-9, CRP levels increase, and higher blood flow shear stress is conducive to maintaining the normal function of vascular endothelium, protecting vascular intima, and preventing hypertension. MicroRNAs (miRNAs) are endogenous non coding RNA small molecules that mainly regulate gene expression at the post transcriptional level. They can inhibit the function of RAAS, reduce water and sodium retention, and regulate the expression of SRF, IRF, GAX and other genes, leading to endothelial dysfunction and participating in the occurrence and development of hypertension by regulating the expression of AT1R gene and mineralocorticoid receptor gene. At the same time, key targets such as PTGS2, JUN, MMP1, MMP9, CXCL2, etc. were significantly enriched in these pathways, fully demonstrating the synergistic antihypertensive effect of mulberry leaf chrysanthemum with multiple targets and pathways.
In conclusion, this paper systematically analyzed the molecular mechanism of mulberry leaf chrysanthemum in the treatment of hypertension by means of network pharmacology and molecular docking, and revealed that mulberry leaf chrysanthemum may act on multiple targets such as JUN, MMP9, PTGS2, EGFR, CCND1, MMP1, PRKCA through IL-17 signaling pathway, fluid shear stress and atherosclerosis signaling pathway The TNF signaling pathway and the microRNA signaling pathway in cancer are involved in vascular relaxation, calcium and sodium ion transport, endothelial function, inflammatory response, glucose and lipid metabolism, and oxidative stress, and play a role in lowering blood pressure. This proves that mulberry leaf chrysanthemum has the characteristics of multi-component, multi-target, and multi-path synergistic effects in the treatment of hypertension, providing direction for further exploration of its pharmacological substance basis and mechanism of action. Further in vitro and in vivo experiments are needed to confirm the intrinsic correlation and target of its efficacy network.