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Based on the holistic regulation and multi-target characteristics of TCM, this study investigated the mechanism of Panlongqi Tablets (PLQT) in ameliorating rheumatoid arthritis (RA) by regulating key proteins, including glycogen synthase kinase-3β (GSK-3β), phosphatidylinositol 3-kinase catalytic subunit alpha (PIK3CA), and protein kinase B3 (AKT3), within the PI3K-AKT signaling pathway. An adjuvant-induced arthritis (AIA) rat model was established by intradermal injection of type Ⅱ collagen-Freund's complete adjuvant into the right hind paw, and rats received different doses of PLQT for intervention. Pharmacodynamic effects were evaluated by monitoring changes in body weight, measuring paw swelling, observing ankle synovial tissue pathology via hematoxylin-eosin (HE) staining, and detecting serum levels of inflammatory cytokines, including tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1β (IL-1β), using ELISA. Targets identified through prior network pharmacology screening were integrated with therapy-related biomarkers from untargeted metabolomics for key pathway enrichment and identification of critical targets. Western blot and RT-qPCR were subsequently used to validate the expression of these key targets. Animal experimental results showed that, compared with the model group, all PLQT dose groups significantly improved weight loss, reduced paw swelling, inhibited synovial tissue fibrosis hyperplasia and inflammatory cell infiltration, and decreased serum levels of pro-inflammatory cytokines such as TNF-α, IL-6, and IL-1β in RA rats, exhibiting a partial dose-dependent manner. Metabolomics analysis revealed that PLQT exerted regulatory effects on 179 of 276 RA-related biomarkers. Enrichment analysis revealed that GSK-3β, PIK3CA, and AKT3 proteins in the PI3K-AKT signaling pathway were the key targets for PLQT intervention in RA. Validation by Western blot and RT-qPCR demonstrated that PLQT significantly modulated the expression of these three proteins. In conclusion, the therapeutic effects of PLQT on RA are likely associated with regulation of GSK-3β, PIK3CA, and AKT3 within the PI3K-AKT signaling pathway.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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