[Effects of Zhuyu Pills on Th1/Th2 and Th17/Treg immune balance in acne model mice via regulation of JAK2/STAT3 signaling pathway].

Based on the Janus kinase 2(JAK2)/signal transducer and activator of transcription 3(STAT3) signaling pathway, this study investigated the effects of Zhuyu Pills on the immune balance of helper T cell(Th) 1/Th2 and Th17/regulatory T cell(Treg) populations in a mouse model of acne. The acne model was established by applying oleic acid to the right auricle combined with subcutaneous injection of Propionibacterium acnes, resulting in auricle lesions of 1 cm~2 over a 14-day modeling period. Fifty successfully modeled ICR mice were randomly assigned to five groups: model, low-, medium-, and high-dose Zhuyu Pills, and isotretinoin(n=10 per group). An additional 10 age-matched healthy ICR mice were included as a control group without modeling. The control and model groups received equal volumes of sterile distilled water via gavage, while the low-, medium-, and high-dose Zhuyu Pills groups received Zhuyu Pills at 130.54, 261.08, and 522.16 mg·kg~(-1), respectively. The isotretinoin group received isotretinoin at 4.55 mg·kg~(-1), once daily. All treatments were administered for four weeks. After administration, auricle tissue pathology was evaluated by hematoxylin and eosin(HE) staining. Serum levels of interleukin(IL)-17, interferon-γ(IFN-γ), IL-10, transforming growth factor-β(TGF-β), IL-12, IL-22, IL-4, and IL-5 were measured by ELISA. Protein expression of p-JAK2/JAK2 and p-STAT3/STAT3 in auricle tissue was assessed by Western blot. Th1, Th2, Th17, and Treg cell populations in the spleen were analyzed by flow cytometry. Immunohistochemistry was used to detect T-box expressed in T cell(T-bet) and GATA binding protein 3(GATA3) expression, and immunofluorescence was used to detect retineic-acid-receptor-related orphan nuclear receptor gamma(RORγt) and forkhead box P3(Foxp3) expression in auricle tissue. Western blot and real-time PCR were used to measure protein and mRNA levels of T-bet, GATA3, RORγt, and Foxp3. Compared with the control group, the model group exhibited significantly thickened auricle epidermis and extensive inflammatory cell infiltration(P<0.01); increased serum IL-17, IFN-γ, IL-12, and IL-22(P<0.01); decreased TGF-β, IL-10, IL-4, and IL-5(P<0.01); elevated p-JAK2/JAK2 and p-STAT3/STAT3 levels in auricle tissue(P<0.01); increased Th1 and Th17(P<0.01) and decreased Th2 and Treg cell populations in the spleen(P<0.01); and upregulated protein and mRNA levels of T-bet and RORγt(P<0.01) but downregulated GATA3 and Foxp3 protein and mRNA levels in auricle tissue(P<0.01). Compared with the model group, all Zhuyu Pills groups and the isotretinoin group showed reduced epidermal thickness and inflammatory infiltration(P<0.01); decreased serum IL-17, IFN-γ, IL-12, and IL-22(P<0.05 or P<0.01) and increased TGF-β, IL-10, IL-4, and IL-5(P<0.05 or P<0.01); lower p-JAK2/JAK2 and p-STAT3/STAT3 levels in auricle tissue(P<0.01); decreased Th1 and Th17(P<0.01) and increased Th2 and Treg cell populations in the spleen(P<0.01); and decreased protein and mRNA levels of T-bet and RORγt(P<0.01) but increased GATA3 and Foxp3 protein and mRNA levels in auricle tissue(P<0.05 or P<0.01). In conclusion, Zhuyu Pills may regulate inflammatory cytokines through the JAK2/STAT3 signaling pathway, restore Th1/Th2 and Th17/Treg polarization balance, maintain immune cell homeostasis, and thereby exert therapeutic effects on inflammatory skin lesions in acne.