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[Xinfeng capsule inhibits immune inflammation in osteoarthritis by inhibiting the miR- 23a-3p/PETN/PI3K/AKT/mTOR
1First Affiliated Hospital of Anhui University of Chinese Medicine, Hefei 230031, China.
Objective:
To observe the inhibitory effect of Xinfeng capsule (XFC) on immune inflammatory response in the coculture system of chondrocytes and CD4+ T cells from patients with osteoarthritis (OA).
Objective:
Thirty OA patients and 30 healthy subjects were examined for the expression of miR-23a-3p/PTEN/PI3K/AKT/mTOR. The changes in the miR-23a-3p/ PTEN/PI3K/AKT/mTOR pathway and the clinical indicators of the OA patients after XFC treatment were observed. OA-CD4+ T cells and OA-Chondrocytes cells were cultured in Transwel chambers, and the expressions of miR-23a-3p, PTEN, PI3K, AKT and mTOR mRNA were detected by RT-PCR; the levels of IL-1β, IL-6, IL-10 and TNF-α were detected by ELISA. The protein expressions of PTEN, PI3K, AKT, p-AKT, mTOR and p-mTOR were detected by Western blotting.
Objective:
Compared with healthy individuals, OA patients showed significantly increased expressions of miR-23a-3p, PTEN, PI3K, AKT, IL-1β, IL-6 and TNF-α and lowered expressions of PTEN and IL-10 (P < 0.01). Positive correlations were found between miR-23a-3p and IL-6 and between PI3K and IL-10; mTOR was positively correlated with IL-6, IL-10, complement C3 and C4 (P < 0.01 or 0.05). Intervention with XFC obviously down-regulated the expressions of IL-1β, IL-6, and TNF-α and up-regulated the expression of IL-10 (P < 0.01). In the cell co-culture systems, the expressions of miR-23a-3p, PI3K, Akt, mTOR, IL-1β, IL-6 and TNF-α were up-regulated while the expressions of PTEN and IL-10 were down-regulated in the model group (P < 0.01). Overexpression of miR-23a-3p significantly up-regulated the expressions of IL-1β, IL-6, TNF-α, miR-23a-3p, PI3K, AKT and mTOR and lowered the expressions of PTEN and IL-10 (P < 0.01 or 0.05). The expressions of IL-1β, IL-6, TNF-α, miR-23a-3p, PI3K, AKT and mTOR were down-regulated and the expressions of IL-10 and PTEN were up-regulated in XFC-treated cells (P < 0.01 or 0.05).
Objective:
XFC can reduce the inflammatory response in patients with OA by down-regulating the expression of miR-23a-3p, inhibiting the activation of the PTEN/PI3K/AKT/ mTOR pathway, and regulating the expression of IL-1β, IL-6, IL-10 and TNF-α.
Insights
Xinfeng capsule (XFC) effectively reduces inflammation in osteoarthritis (OA) by modulating immune responses. It targets the miR-23a-3p/PTEN/PI3K/AKT/mTOR pathway, decreasing pro-inflammatory cytokines and increasing anti-inflammatory ones.
Area of Science:
- Immunology and Molecular Biology
- Osteoarthritis Pathogenesis Research
- Traditional Chinese Medicine Pharmacology
Background:
- Osteoarthritis (OA) is characterized by immune-mediated inflammation involving chondrocytes and CD4+ T cells.
- The miR-23a-3p/PTEN/PI3K/AKT/mTOR pathway plays a critical role in regulating inflammatory responses in OA.
- Understanding the molecular mechanisms underlying OA inflammation is crucial for developing effective treatments.
Purpose of the Study:
- To investigate the inhibitory effect of Xinfeng capsule (XFC) on the immune inflammatory response in an OA coculture system.
- To analyze the impact of XFC on the miR-23a-3p/PTEN/PI3K/AKT/mTOR pathway and related cytokines in OA patients.
- To elucidate the therapeutic potential of XFC in managing OA-associated inflammation.
Main Methods:
- Coculture of chondrocytes and CD4+ T cells from OA patients and healthy individuals.
- Quantification of miR-23a-3p, PTEN, PI3K, AKT, and mTOR mRNA expression using RT-PCR.
- Measurement of IL-1β, IL-6, IL-10, and TNF-α levels via ELISA and protein expression via Western blotting.
Main Results:
- OA patients exhibited elevated miR-23a-3p, PI3K, AKT, IL-1β, IL-6, and TNF-α, with decreased PTEN and IL-10 compared to healthy controls.
- XFC treatment significantly down-regulated IL-1β, IL-6, TNF-α, and miR-23a-3p, while up-regulating IL-10 in OA cell models.
- XFC inhibited the PTEN/PI3K/AKT/mTOR pathway activation, correlating with reduced inflammatory markers.
Conclusions:
- Xinfeng capsule demonstrates significant anti-inflammatory effects in osteoarthritis by modulating immune cell interactions.
- XFC exerts its therapeutic action through the downregulation of miR-23a-3p and inhibition of the PTEN/PI3K/AKT/mTOR signaling pathway.
- XFC represents a promising therapeutic agent for osteoarthritis, targeting key inflammatory mediators and pathways.
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