Exploring the effective ingredients and potential mechanisms of action of Da Xue Teng in treating cerebral infarction based on ultra-high performance liquid chromatography-mass spectrometry technology and network pharmacology
Stroke is the leading cause of reduced life expectancy among Chinese citizens, seriously endangering their health and affecting their overall health level. With the development of social economy and the improvement of national living standard, the incidence rate of stroke is increasing year by year and is becoming younger. According to the different types of onset, stroke is divided into ischemic stroke and hemorrhagic stroke. Ischemic stroke, also known as cerebral infarction, is reported to account for approximately 85% of the total stroke population in China. Therefore, how to reduce the incidence rate, disability rate, mortality rate, and restore neurological function of cerebral infarction is a difficult and hot topic for Chinese and Western medicine researchers.
At present, the main treatment methods for cerebral infarction in Western medicine are intravenous thrombolysis, arterial thrombolysis, and mechanical thrombectomy, which can make positive contributions to the short-term reconstruction of lesion blood supply, alleviation of hypoxia damage, and reduction of free radical production for patients. But some patients have poor long-term prognosis and face problems such as sequelae and complications. Traditional Chinese medicine has its unique advantages in treating cerebral infarction, including a wide range of applications, minimal side effects, good patient prognosis, improved neurological function recovery, reduced disability and recurrence rates, and improved patient quality of life. Traditional Chinese medicine believes that the etiology and pathogenesis of cerebral infarction are related to qi deficiency and blood stasis, phlegm and blood stasis obstructing the orifices, etc. With the deepening understanding and research of stroke and “toxic pathogens”, some scholars have proposed viewpoints such as “heat toxicity theory of stroke” and “toxic damage to the brain meridian”, believing that phlegm toxicity, heat toxicity, etc. produced after stroke can all damage the brain meridian. The big blood vine is Sargentodoxa cuneta (Oliv.) Rehd, a member of the family Actinaceae The dried vine stems of Et Wils plants have the effects of clearing heat and detoxifying, promoting blood circulation, dispelling wind and dampness, and contain rich chemical components such as saponins, anthraquinones, organic acids, lignin, phenols, etc. Commonly used for treating intestinal abscess, abdominal pain, heat toxin sores, amenorrhea, dysmenorrhea, swelling and pain, rheumatism and rheumatism. Modern pharmacological studies have shown that Scutellaria baicalensis can improve damage to the hippocampus and cortex of the brain, inhibit the decrease in microcirculatory perfusion, reduce the proportion of infarcted areas, and lower mortality rates. The total phenolic acid of Scutellaria baicalensis can inhibit the inflammatory response of brain tissue and apoptosis of brain cells in rats with cerebral ischemia-reperfusion injury. At the same time, it can reduce the mortality rate and neurological deficit score of rats with cerebral ischemia-reperfusion injury. Tongqiao Huashuan Formula is an effective formula for clinical treatment of cerebral infarction, among which Da Xue Teng is the main medicine. The core functions of this formula are to dispel wind, phlegm, and blood stasis. The clinical research results of Cai et al. showed that Tongqiao Huashuan Tang can significantly improve the hemorheological indicators of ischemic stroke patients, improve blood viscosity, coagulation, and cell aggregation, and has ideal antithrombotic effects. At present, the effective ingredients, targets, and pathways of action of Tripterygium wilfordii on cardiovascular and cerebrovascular diseases are still in the exploratory stage, which limits the research and development, utilization, and clinical promotion. Therefore, based on previous research, this study intends to further analyze the chemical composition of Scutellaria baicalensis, clarify the therapeutic targets and pathways of its active ingredients on cerebral infarction, and provide scientific basis for further research on the treatment of cerebral infarction with Scutellaria baicalensis.

Analyze the composition of the top 5 components with the highest degree values in the constructed network graph, including dihydroguaiacol, emodin, ionoside, carotenoids, and emodin methyl ether. Dihydroguaiacol has a blocking effect on platelet activating factor receptor binding, thereby preventing platelet aggregation and thrombus formation. Rhubarb may exert neuroprotective effects by inhibiting the expression level of Bax protein, reducing the production of Beclin1 protein, and alleviating neuronal apoptosis. The neuroprotective effect of carotenoids is related to the upregulation of antioxidant proteins, inhibition of P38 mitogen activated protease, and activation of stress activated protein kinase signaling pathways. In addition, studies have found that carotenoids have a promoting effect on the proliferation of neural stem cells. Emodin methyl ether can reduce the expression of IL-1 β, TNF – α, and ICAM-1, inhibit the activity of Caspase-3, thereby reducing neuronal apoptosis and exerting neuroprotective effects.
Based on the selected 71 core targets, literature analysis was conducted on targets with high degree values such as PIK3CA, Src, STAT3, JAK2, etc. PIK3CA is a central molecule in the PI3K/AKT/mTOR pathway, which is associated with cell proliferation, angiogenesis, and tumorigenesis. Studies have shown that the expression of PIK3CA protein is significantly inhibited in SH-SY5Y (human neuroblastoma cells) and brain tissue induced by OGD/R (oxygen glucose deprivation/reoxygenation). Activation of the PI3K/Akt/mTOR signaling pathway can effectively increase the expression level of PIK3CA, thereby reducing mitochondrial mediated cell apoptosis and exerting neuroprotective effects. Astrocytes (Src) are the most abundant cells in the central nervous system. After cerebral infarction, they can be activated to become reactive astrocytes and form glial scars. Studies have shown that Src is an upstream signal for AngII induced activation of astrocytes, and Src can activate ERK1/2 to stimulate the growth of astrocytes, playing an important role in the process of brain injury. The activation of JAK2/STAT3 is closely related to neuronal apoptosis. Studies have shown that JAK2, as an upstream signaling molecule of STAT3, can regulate the phosphorylation of STAT3, thereby mediating downstream biological responses to STAT3 activation. JAK phosphorylation inhibitors can reduce the phosphorylation levels of JAK2 and STAT3 in ischemic brain tissue, thereby lowering the apoptosis rate and reducing the infarct size. HSPs are stress proteins produced by cells upon exposure to harmful stimuli, which are associated with the generation, proliferation, and differentiation of tumors. There are research reports that HSP90 α can protect hypoxic nerve cells to a certain extent and improve their ability to withstand hypoxia, buying time for vascular regeneration in the penumbra. In summary, the predicted core targets are involved in the pathogenesis of cerebral infarction, and the analysis results are consistent with relevant reports on Scutellaria baicalensis and cerebral infarction. It is speculated that the chemical components of Scutellaria baicalensis can exert potential therapeutic mechanisms by acting on these core targets.
According to the KEGG enrichment analysis results, it can be concluded that Scutellaria baicalensis may play a role in preventing and treating cerebral infarction through the HIF-1 signaling pathway, Sphingolipid signaling pathway, ErbB signaling pathway, and other pathways. The HIF-1 signaling pathway is the main molecule regulating cells’ response to hypoxia, and it is more sensitive during cellular hypoxia. At present, it has been found that the HIF pathway can directly stimulate and activate nearly 80 genes, and the induced protein expression is mainly manifested in improving the tolerance and survival of hypoxic cells. Sphingolipids play an important role in the process of nerve growth and differentiation. Exogenous sphingolipids have been proven to have a wide range of neuroprotective effects, such as brain protection and promoting the repair of damaged nerves. There are studies showing that the combination of intravenous thrombolysis and sphingolipids injection has better neurological function recovery than single line intravenous thrombolysis. During the aging process and the development of neurodegenerative diseases, the levels of ceramides in the brain gradually increase and mediate neuronal apoptosis, playing a repairing role. The NRG/ErbB signaling pathway plays an important role in the protective mechanism of cerebral infarction. Studies have found that in traumatized brain tissue, NEG-1 β and ErbB receptors in nerve cells are induced. ErbB signaling can antagonize the production of central nervous system inflammation and protect brain neurons. Research has also found that acupuncture can regulate the NRG/ErbB pathway, increase the expression level of ErbB, and to some extent inhibit neuronal apoptosis and the release and expression of inflammatory factors. It can also inhibit monocyte infiltration, activation of astrocytes, and cytokine production, thereby exerting a protective effect on neuronal cells by suppressing inflammatory responses. The above-mentioned pathways mainly participate in processes such as oxidative stress, neural repair, cell apoptosis, and inflammatory response. It can be further speculated that the chemical components of Scutellaria baicalensis can exert a therapeutic mechanism for cerebral infarction by regulating these pathways. In addition, the results of molecular docking preliminarily simulated the interaction relationship between components and targets, which to some extent verified the reliability of the predicted results.
In summary, this study combined UPLC-HR-MS, network pharmacology, molecular docking, and other methods to investigate and analyze the chemical components of Scutellaria baicalensis and its therapeutic targets and biological pathways for cerebral infarction. The interaction relationship between chemical components and targets was constructed and validated. The mechanism of action of Tripterygium wilfordii in treating cerebral infarction through multi-target and multi pathway mechanisms has been elucidated from multiple perspectives, and the relevant predictions are consistent with literature reports, providing a basis and evidence for further research and development of the mechanism of action of Tripterygium wilfordii in treating cerebral infarction in the future.