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Background: Acute gouty arthritis (AGA) significantly impairs patients' quality of life. Currently, existing therapeutic agents exhibit definite efficacy but also lead to serious adverse reactions. Therefore, it is essential to develop highly efficient therapeutic agents with minimal adverse reactions, especially within traditional Chinese medicine (TCM). Additionally, food polyphenols have shown potential in treating various inflammatory diseases. The Qingre-Huazhuo-Jiangsuan-Recipe (QHJR), a modification of Si-Miao-San (SMS), has emerged as a TCM remedy for AGA with no reported side effects. Recent research has also highlighted a strong genetic link to gout. Methods: The TCM System Pharmacology (TCMSP) database was used to collect the main chemical components of QHJR and AGA-related targets for predicting the metabolites in QHJR. HPLC-Q-Orbitrap-MS was employed to identify the ingredients of QHJR. The collected metabolites were then used to construct a Drugs-Targets Network in Cytoscape software, ranked based on their "Degree" of significance. Differentially expressed genes (DEGs) were screened in the Gene Expression Omnibus (GEO) database using GEO2R online analysis. Subsequently, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed. The DEGs were utilized to construct a Protein-Protein Interaction (PPI) Network via the STRING database. In vivo experimental validation was conducted using colchicine, QHJR, rapamycin (RAPA), and 3-methyladenine (3-MA) as controls to observe QHJR's efficacy in AGA. Synovial tissues from rats were collected, and qRT-PCR and Western blot assays were employed to investigate Ampk-related factors (Ampk, mTOR, ULK1), autophagy-related factors (Atg5, Atg7, LC3, p62), and inflammatory-related factors (NLRP3). ELISA assays were performed to measure inflammatory-related factor levels (IL-6, IL-1ß, TNF-α), and H&E staining was used to examine tissue histology. Results: Network analysis screened out a total of 94 metabolites in QHJR for AGA. HPLC-Q-Orbitrap-MS analysis identified 27 of these metabolites. Notably, five metabolites (Neochlorogenic acid, Caffeic acid, Berberine, Isoliquiritigenin, Formononetin) were not associated with any individual herbal component of QHJR in TCMSP database, while six metabolites (quercetin, luteolin, formononetin, naringenin, taxifolin, diosgenin) overlapped with the predicted results from the previous network analysis. Further network analysis highlighted key components, such as Caffeic acid, cis-resveratrol, Apigenin, and Isoliquiritigenin. Other studies have found that their treatment of AGA is achieved through reducing inflammation, consistent with this study, laying the foundation for the mechanism study of QHJR against AGA. PPI analysis identified TNF, IL-6, and IL-1ß as hub genes. GO and KEGG analyses indicated that anti-inflammation was a key mechanism in AGA treatment. All methods demonstrated that inflammatory expression increased in the Model group but was reversed by QHJR. Additionally, autophagy-related expression increased following QHJR treatment. The study suggested that AMPKα and p-AMPKα1 proteins were insensitive to 3 MA and RAPA, implying that AMPK may not activate autophagy directly but through ULK1 and mTOR. Conclusion: In conclusion, this study confirms the effectiveness of QHJR, a modified formulation of SMS (a classic traditional Chinese medicine prescription for treating gout), against AGA. QHJR, as a TCM formula, offers advantages such as minimal safety concerns and potential long-term use. The study suggests that the mechanism by which QHJR treats AGA may involve the activation of the AMPK/mTOR/ULK1 pathway, thereby regulating autophagy levels, reducing inflammation, and alleviating AGA. These findings provide new therapeutic approaches and ideas for the clinical treatment of AGA.
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ETHNOPHARMACOLOGICAL RELEVANCE: Gout belongs to the category of "arthralgia syndrome" in traditional Chinese medicine. It is believed that gout is caused by stagnation of blood stasis, heat, and turbid toxin. Qingre Huazhuo Jiangsuan Decoction (QHJD) is a traditional Chinese medicine prescription developed from the classic Chinese medicine prescription Simiao powder to clear heat, remove turbidity, reduce acid, and reduce inflammation. Now Traditional Chinese Medicine (TCM) physicians often apply it to treat acute gouty arthritis (AGA). However, the mechanism of QHJD in relieving acute gouty arthritis is still unclear, and further research is needed. AIM OF THE STUDY: Here, we aim to explore the potential mechanism of QHJD in relieving acute gouty arthritis. MATERIALS AND METHODS: Acute gouty arthritis model was established by injecting monosodium urate (MSU) suspension into knee joint. The pathological state of synovial tissue in each group was evaluated by hematoxylin-eosin (HE) staining. The level of TNF-α, IL-6, and IL-1ß were detected by enzyme-linked immunosorbent assay (ELISA). qRT-PCR was used to detect the mRNA expression of NLRP3, ATG5, ATG7, PI3K, AKT, and mTOR. The protein expression of LC3II/I, p62, ULK1, P-ULK1, Beclin-1, PI3K, AKT, mTOR, P-PI3K, P-AKT, and P-mTOR were detected by Western blot. RESULTS: (1) The level of autophagy protein (mRNA) was significantly up-regulated in QHJD group and rapamycin, while the expression of autophagy protein (mRNA) was significantly downregulated in the 3-methyladenoenoic acid (3 MA) group; (2) QHJD and rapamycin significantly inhibited PI3K/AKT/mTOR pathway, while 3 MA group activated this pathway. (3) It was worth noting that after treatment with QHJD and rapamycin, the inflammatory pathological state of AGA synovial tissue was significantly reduced with the activation of the autophagy gene in knee synovial tissue, and the inhibition of PI3K/AKT/mTOR pathway. CONCLUSIONS: This research revealed that QHJD activates autophagy by inhibiting PI3K/AKT/mTOR pathway, thereby relieving acute gouty arthritis.
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Artritis Gotosa , Proteínas Proto-Oncogénicas c-akt , Humanos , Proteínas Proto-Oncogénicas c-akt/metabolismo , Fosfatidilinositol 3-Quinasas/metabolismo , Artritis Gotosa/inducido químicamente , Artritis Gotosa/tratamiento farmacológico , Artritis Gotosa/metabolismo , Serina-Treonina Quinasas TOR/metabolismo , Transducción de Señal , Autofagia , Sirolimus , ARN MensajeroRESUMEN
Lung cancer is one of the most common malignant tumors in the world, and its incidence and mortality rate rank among the top malignant tumors worldwide, which has become an important killer threatening human survival rate and well-being. Modern medical treatment for lung cancer is mainly based on surgery and radiotherapy, with gene, targeted drugs and immunotherapy as auxiliary treatments, which are effective, but there are problems such as postoperative recurrence, resistance to radiotherapy, toxic side effects and poor compliance. In recent years, with the continuous development of TCM, TCM is popular among physicians and patients for its high efficiency, low toxicity, low side effects and economic benefits, etc. As a classical TCM formula, Qianjin Weijin Decoction(QJWJ) has certain value in the treatment of lung cancer. This paper summarizes and analyzes the clinical research, molecular mechanism, pharmacological effects and chemical composition of QJWJ in the treatment of lung cancer, in order to provide more ideas and theoretical basis for the treatment of lung cancer with QJWJ.