Study on the mechanism of Huangqi extract in treating ischemic hypoxic encephalopathy based on network pharmacology, molecular docking technology, and experimental verification
Neonatal ischemic encephalopathy (HIE) is a type of disease that affects the nerve growth and development of newborns and infants. Survivors often leave behind various neurological sequelae, which impose a heavy burden on the affected children and their families. There is currently no specific drug for treating HIE in clinical practice, and studying the pathophysiological mechanisms of HIE is of great significance for finding more effective treatment plans. Research has shown that oxidative stress, calcium influx, free radical generation, acidosis, ion imbalance, inflammation, apoptosis, autophagy, necrosis, and other factors all play important roles in the pathogenesis of HIE. In the field of traditional Chinese medicine, it has been reported that Huangqi and its preparations have shown good effects in the treatment of HIE. Modern pharmacology has confirmed that Huangqi extract has the effects of clearing oxygen free radicals, anti infection, and protecting vascular endothelium. It is widely used in clinical practice to treat cardiovascular and cerebrovascular diseases. This study aims to analyze the mechanism of action of Huangqi extract in treating HIE through network pharmacology and molecular docking techniques, providing reference for promoting clinical application.

The main pathophysiological mechanism of HIE is ischemia and hypoxia of brain tissue. Brain tissue ischemia and hypoxia can cause a series of pathological and physiological changes, mainly including cell apoptosis, inflammatory response, and oxidative stress response. The different stages of ischemic and hypoxic nerve damage are all caused by the interaction between the three physiological changes mentioned above. The treatment of neonatal HIE has always been a hot and difficult topic in clinical research. The traditional “three supports” and “three symptomatic” therapies are limited by various factors such as gestational age, time, and medical facilities, resulting in limited therapeutic effects for HIE. Clinically, there has been an urgent search for complementary or alternative therapies for the treatment of HIE. Huangqi, as a traditional Chinese medicine, has made significant progress in recent years through modern pharmacological research on the effects of active ingredients in Huangqi. Research has shown that Astragalus membranaceus has the effects of enhancing the body’s immunity, resisting hypoxia and oxidative stress, and improving vascular endothelial function. Animal experiments have shown that Astragalus membranaceus can upregulate the expression level of B-lymphotumor-2 protein in rat brain tissue, inhibit neuronal apoptosis, and alleviate brain damage caused by ischemia and hypoxia. However, the main active ingredients and mechanisms of action of Astragalus membranaceus in clinical practice are not yet clear. Network pharmacology and molecular docking are emerging technologies in recent years for pharmacological research based on large databases. Through big data, the main active ingredients and targets of related drugs are obtained and molecular docking is performed with disease targets to analyze the specific mechanism of action of drugs. According to the TCMSP database, a total of 70 active ingredients of Huangqi extract were obtained. A total of 350 relevant targets were obtained, and after removing duplicate targets and gene annotations, 120 targets of the active ingredients in “Huangqi Extract” were finally obtained.
The compound network target map obtained through topological analysis shows that the main active ingredients of Astragalus membranaceus in treating HIE are 4-hydroxycinnamic acid, cis avionic acid, astragaloside, hederagenin, and galangin. 4-hydroxycinnamic acid has been proven to have the ability to scavenge oxygen free radicals. Animal experiments have shown that 4-hydroxycinnamic acid can alleviate neuropathic pain in rats. A study in Xizang showed that cis acitretin can effectively improve the oxygen saturation of the body and alleviate the adverse symptoms caused by hypoxia; And the active ingredient can promote the absorption of astragaloside IV in the body. Astragaloside IV has been extensively studied and proven to have anti-inflammatory, anti apoptotic, and immunomodulatory biological activities, making it an important neuroprotective agent. Astragaloside rosewood glycoside is a compound isolated from Astragalus membranaceus, which can inhibit the activation of extracellular regulated protein kinases 1/2 (ERK1/2) induced by human platelet-derived growth factor BB (PDGF-BB) stimulation, and inhibit the phosphorylation of mitogen activating protein (MAP) kinase; It can also inhibit PDGF-BB-induced proliferation of vascular smooth muscle cells by suppressing the ERK1/2MAP kinase cascade pathway. Network pharmacology studies have shown that the active ingredient of Huangqi, Hualiangjiangsu, can exert therapeutic effects on knee osteoarthritis through the PDK Akt signaling pathway, tumor necrosis factor, and MAPK signaling pathway.
After mapping 120 targets with 242 targets of HIE, 18 potential targets were obtained. Further PPI network analysis revealed that NOS2, PTGS2, SOD1, MAPK4, and CASP8 are the core targets of Huangqi in treating HIE. Inflammation is considered an important pathogenic factor of HIE, and a large amount of inflammatory factors can accelerate the apoptosis and necrosis of neuronal cells. NOS2 can promote the production of nitric oxide in the neural tissues of the central and peripheral nervous systems, and participate in inflammation and oxidative stress responses. Animal experiments have shown that knocking out the rat NOS2 gene can inhibit the synthesis of Rac1, thereby suppressing the pro-inflammatory effect of tumor necrosis factor alpha. The main function of SOD1 is to exert antioxidant effects in the body. SOD1 plays an important role in the oxidative stress response of the body, as it can promote the breakdown of hydrogen peroxide in cells, reduce the production of oxygen free radicals, and maintain the balance of the body’s oxidative antioxidant system. In vitro cell experiments have shown that overexpression of PTGS2 can exacerbate cell damage in PC12 cells under glucose deprivation model. When local ischemia and hypoxia occur and release cytokines, it can activate the MAPK4 pathway, further stimulating immune cells to release pro-inflammatory cytokines and exacerbating local inflammatory reactions. CASP8 is a key regulatory factor for cell apoptosis and necrotic apoptosis, as well as a key factor in regulating inflammatory response through its dimerization and enzyme activity. An increase in its content indicates an enhancement of local inflammatory response. From the above analysis, we can see that the main active ingredients of Astragalus membranaceus exert their therapeutic effects on HIE by acting on the above-mentioned targets, and its biological processes may involve inflammatory reactions, oxidative stress cells, damage repair, etc.
The molecular docking results showed that the three active ingredients in Astragalus membranaceus, 4-hydroxycinnamic acid, cis avionic acid, and astragaloside, have good binding activity with key targets, mainly relying on hydrophobic or hydrogen bonding to exert therapeutic effects on HIE. In terms of binding energy, the lowest binding energy between SOD1 and Astragalus membranaceus rosewood glycoside is -8.9 kcal/mol. The animal experiment verification results showed that the expression levels of NOS2 and PTGS2 proteins in the hippocampus of the treatment group rats were lower than those in the model group, and the expression level of SOD1 protein was higher than that in the model group. The expression levels of NOS2 and PTGS2 proteins in the hippocampus of the sham surgery group rats were lower than those in the treatment group and the model group, and the expression level of SOD1 was higher than that in the treatment group and the model group, further confirming the reliable role of Huangqi extract in the treatment of HIE. Related studies have shown that astragalus saponins can activate the Nrf2/HO signaling pathway, upregulate the expression of downstream antioxidant gene HO-1, inhibit nitric oxide production, increase SOD activity, and enhance the antioxidant capacity of brain tissue. Modern pharmacology has also confirmed that besides its strong ability to scavenge free radicals, Huangqi contains flavonoids that can reduce platelet activity during cerebral ischemia-reperfusion, inhibit the expression of cyclooxygenase-2, and alleviate cerebral ischemia-reperfusion injury.
In summary, this article uses network pharmacology and molecular docking technology to analyze the mechanism of the main active ingredients in Astragalus membranaceus extract in treating HIE. It is concluded that the three main components of Astragalus membranaceus extract in treating HIE are 4-hydroxycinnamic acid, cis avionic acid, and Astragalus membranaceus rosewood glycoside, which act on key targets such as NOS2, PTGS2, SOD1, MAPK4, and CASP8 to exert their therapeutic effects on HIE. Network pharmacology analysis also confirmed that this process is the result of a synergistic effect of multiple components, targets, and pathways.