Research on the medication patterns and molecular mechanisms of high-frequency drug combinations in ankylosing spondylitis based on data mining and network pharmacology

Research on the medication patterns and molecular mechanisms of high-frequency drug combinations in ankylosing spondylitis based on data mining and network pharmacology
Ankylosing spondylitis (AS) is a chronic inflammatory disease of unknown etiology. Unlike other systemic autoimmune diseases, the abnormal activity of the innate immune system and immune cells plays a dominant role. During the development of the disease, AS is prone to involve the sacroiliac joint and axial bone area, with the most common symptoms being chronic back pain and spinal stiffness, but peripheral and musculoskeletal manifestations are also common. In terms of treatment, nonsteroidal anti-inflammatory drugs (NSAIDs) remain the first-line drug therapy. As second-line drugs, tumor necrosis factor (TNF) inhibitors and interleukin-17 (IL17) inhibitors are available. Recently, Janus kinase inhibitors have also shown efficacy in improving disease symptoms. Although these biologics have shown good efficacy in the treatment of AS, there are also many adverse reactions. Traditional Chinese medicine has been passed down for thousands of years and has its own unique therapeutic advantages. Traditional Chinese medicine experts have been tirelessly pursuing the treatment of AS, and traditional Chinese medicine has unique advantages in improving AS symptoms and enhancing long-term efficacy. Shanghai University of Traditional Chinese Medicine Affiliated Guanghua Hospital is a tertiary specialized hospital for joint diseases, with an annual outpatient volume of over 230000 and an annual discharge volume of over 6800. It has 153 beds and is one of the largest rheumatology departments in China, with certain research advantages. Due to the fact that traditional Chinese medicine is an empirical medicine with multiple schools of thought, there is no unified standard for the use of medication in the treatment of AS diseases. Traditional Chinese medicine prescriptions are mostly self formulated and lack modern pharmacological research. Therefore, the author adopts data mining techniques to explore the prescription and medication rules of rheumatology and immunology physicians in the hospital using traditional Chinese medicine to treat AS, analyzes the high-frequency use of single herbs, drug pairs, and the four qi and five flavor, efficacy classification, and meridian rules of high-frequency drugs, and excavates new prescriptions. At the same time, combined with traditional Chinese medicine pharmacology analysis, the molecular mechanism of traditional Chinese medicine in treating AS is further explained, providing a certain theoretical basis and guidance for the treatment of AS diseases with traditional Chinese medicine.

 

The etiology and pathogenesis of AS cannot be fully explained at present. Nowadays, traditional Chinese medicine generally believes that in the early stage, this disease is mainly characterized by pathogenic factors. Over time, phlegm and blood stasis accumulate, blocking the Du meridian and causing stiffness and discomfort in the spine. In the later stage, liver and kidney deficiency and damage to the Du meridian occur, forming the pathological characteristics of “early pathogenic factors, middle blood stasis, and late deficiency”. Traditional Chinese medicine has a long history of treating AS, and the treatment prescriptions of ancient doctors are also very rich. With the development of modern information technology and the use of online databases, we can discover the medication patterns of traditional Chinese medicine in treating diseases, and summarize the medicinal properties, meridian tropism, and efficacy of traditional Chinese medicine, further studying diseases from the perspective of traditional Chinese medicine.

The first part of the article collects and organizes traditional Chinese medicine prescriptions for treating AS in our hospital through data mining. A total of 200 prescriptions were collected, and 236 traditional Chinese medicines were counted. The results showed that modern doctors used drugs similar to those used by ancient doctors in the treatment of AS. In terms of medication, the main drugs used are tonics for tonifying deficiency, promoting blood circulation and removing blood stasis, clearing heat, and dispelling wind and dampness, such as dogspine dispelling rheumatism, tonifying liver and kidney, strengthening muscles and bones, continuing to nourish liver and kidney, Danggui promoting blood circulation and removing blood stasis, Duhuo dispelling wind and dampness, etc. The main flavors of medicine are bitter, sweet, and spicy, while the main medicinal properties are warm, calm, and cold. It travels through meridians such as the liver, lungs, spleen, stomach, and kidneys. Traditional Chinese medicine theory holds that the liver controls the tendons, the kidneys control the bones and store essence, and the spleen and stomach control the circulation and transformation, which are the foundation of postnatal development. If the liver, kidney, spleen, and stomach functions are normal, then the essence and blood are sufficient, and the muscles and bones can be nourished before the spinal joints can move normally. Afterwards, cluster analysis was conducted on high-frequency drugs with a usage frequency of ≥ 1%. The results are divided into six categories, among which the main ones are rheumatism dispelling, blood activating and stasis removing, and kidney tonifying and bone strengthening drugs. This is consistent with the pathological characteristics of AS, which is often characterized by liver and kidney deficiency, damage to the Du meridian, and accompanied by chronic stagnation and stasis. The analysis of association rules shows that the 33 drug combinations with high rankings are mainly composed of tonifying deficiency drugs, promoting blood circulation and removing blood stasis drugs, and dispelling wind and dampness drugs. Examples such as Gouji Xuduan, Duhuo Gouji, Chuanxiong Danggui, etc. can correspond to the high-frequency analysis results of traditional Chinese medicine, confirming the reliability of the data analysis results. In the factor analysis section of traditional Chinese medicine, 8 common factors were extracted, such as CF1 being a combination of heat clearing drugs, blood activating and stasis removing drugs, and wind and dampness removing drugs, CF2 being a combination of wind and dampness removing drugs, deficiency tonifying drugs, and blood activating and stasis removing drugs, and CF3 being a combination of wind and dampness removing drugs and deficiency tonifying drugs. It can be seen that the results of factor analysis and association rule analysis are basically consistent, with high accuracy.

The second part further conducts network pharmacology analysis on the high-frequency drug combination Gouji Xuduan discovered in the data mining results, and elaborates on the specific molecular mechanism of Gouji Xuduan in treating AS. The analysis of drug components found that the active ingredients that play a major role in the treatment of AS with dogspine dipsacus are β – sitosterol, kaempferol, and naringenin, which have important pharmacological effects. β – sitosterol is one of the plant sterols and belongs to the tetracyclic triterpenoid class. β – sitosterol is widely used in the pharmaceutical industry due to its unique biological and physicochemical properties. Research has found that β – sitosterol has important anti-inflammatory, antioxidant, anti-tumor, wound healing promoting, and immune regulatory effects. It inhibits the aggregation and release of tumor necrosis factor and is mainly used for the treatment of cardiovascular and tumor diseases. In addition, studies have found that kaempferol also has anti-inflammatory, anti bacterial and viral infection, and immune regulatory effects. It can enhance the inhibitory function of regulatory T cells, increase the expression level of FOXP3, and prevent and treat inflammatory diseases such as AS. Through pharmacological analysis, it can be found that there are multiple active ingredients with anti-inflammatory, antioxidant, and immune regulatory effects in dogspine dipsacus, which are the key components for dogspine dipsacus to exert therapeutic effects on AS. Each drug component has its own target protein. To further elucidate the relationship between the two, network analysis was conducted between the components and genes, resulting in the identification of multiple major proteins such as TNF, EGFR, VEGFA, STAT3, HSP90AA1, PTGS2, MMP2, MMP9, IL2, etc. There have been many reports on the mechanism of action of TNF in autoimmune and inflammatory diseases, and related therapeutic drugs have also been applied in clinical treatment with good efficacy. PTGS2 is a key enzyme in prostaglandin biosynthesis, serving as both a dioxygenase and a peroxidase. It often participates in various inflammatory reactions of the body and has the functions of inhibiting the secretion of inflammatory factors, anti-inflammatory, and analgesic effects. In addition, there have been many studies on genes such as EGFR and STAT3, which will not be elaborated in detail. Based on the above analysis, it can be found that genes such as TNF, EGFR, VEGFA, STAT3, HSP90AA1, PTGS2, etc. are key genes involved in the action of dog spine on AS. Subsequent pathway enrichment analysis revealed that these key genes are mainly enriched in Toll like receptor signaling pathways and T cell receptor signaling pathways, and are mainly involved in processes such as inflammation regulation, nuclear receptor activity, and ligand activated transcription factor activity. Research has shown that Toll like receptors are important initiators of immune responses, including innate and acquired immune responses. The genetic polymorphism in Toll like receptors can lead to dysfunction of certain related key signaling pathways, thereby increasing the risk of autoimmune diseases, such as regulating the inflammatory response of AS. T cells also play an important role in immune response, as they can help generate protective immunity by eliminating infections and tumors, or regulate immune and inflammatory responses by disrupting host cells. By precisely targeting antigen-specific T cell receptor signals to regulate T cell responses, it is possible to fundamentally change the treatment strategies for autoimmune diseases. In summary, the important active ingredients such as β – sitosterol and kaempferol in Dogspine Dipsacus can act on Toll like receptor signaling and T cell receptor signaling pathways through genes such as TNF, EGFR, and PTGS2 to treat AS. This reflects the characteristics of traditional Chinese medicine in treating diseases with multiple components, targets, and pathways, and provides a better theoretical basis for the application of data mining results.

The article summarizes and analyzes the prescription and medication patterns of traditional Chinese medicine treatment for AS at Guanghua Hospital affiliated with Shanghai University of Chinese Medicine through the application of data mining and network pharmacology methods, and discovers commonly used drug combinations such as Gouji Xuduan. And the molecular mechanism of action of this drug combination was studied, intuitively verifying the characteristics of the multi-component, multi-target, and multi-path treatment of AS with Gouji Xuduan, reflecting the unique advantages of traditional Chinese medicine in treating diseases. In addition, this study also has some shortcomings, as some doctors prescribe traditional Chinese medicine strictly according to the agreed prescription, which limits the collected prescription data to a certain extent. However, network pharmacology analysis only theoretically elucidates the molecular mechanism of action of Gouji Xuduan in treating AS, and further animal or cell experiments are needed to verify and improve the research.