Therapeutic effects of Jieduquyuziyin prescription on the balance of Th17/Treg cells via AMPK/mTOR/HIF-1α signaling pathway in systemic lupus erythematosus

Purpose: This study aims to investigate the capacity of Jieduquyuziyin Prescription (JP) to ameliorate Systemic lupus erythematosus (SLE)-like symptoms and decipher the molecular mechanisms governing JP-mediated re-equilibration of Th17/Treg cells. Methods: An imiquimod (IMQ)-induced murine model of SLE was established. The mice were randomized to receive JP, prednisone (Pred) or metformin (Met) for a 4-week intervention period. Systemic involvement was assessed by monitoring body weight fluctuations and proteinuria levels, while hepatorenal functions were quantified through serum biochemical profiles. Serum concentrations of anti-dsDNA (IgG), anti-Smith antibodies, TGF-β, and IL-17 were determined using ELISA method. Histopathological alterations and immune complex depositions in renal tissues were characterized by H&E and immunofluorescence staining, respectively. Flow cytometry was utilized to quantify the Th17 (CD4+IL-17+) and Treg (CD4+CD25+Foxp3+) cell populations within splenic single-cell suspensions. Furthermore, the splenic expression of lineage-specific transcription factors RORγt and Foxp3, along with the regulatory dynamics of the AMPK/mTOR/HIF-1α signaling, were analyzed using RT-qPCR and western blotting. Results: JP intervention exerted potent systemic therapeutic effects. JP markedly suppressed the titers of key pathogenic autoantibodies (specifically anti-dsDNA and anti-Smith) and improved the renal pathology and the deposition of immune complex IgM in the kidney. Pharmacological intervention shifted the immune balance by attenuating serum IL-17A and promoting TGF-β1, biochemical changes that correlated with the reciprocal modulation of splenic Foxp3 and RORγt mRNA levels. JP markedly augmented splenic Treg frequency while attenuating the Th17 cell response, thereby restoring immune homeostasis. Moreover, JP promoted the activation of the AMPK/CPT-1. Meanwhile, JP decreased the mRNA levels of GLUT1, HK2, PKM2, CPT-1. Conclusion: JP restores Th17/Treg balance via the AMPK/mTOR/HIF-1α pathway, thereby alleviating SLE-like pathogenesis.