A Novel Drug Combination of Mangiferin and Cinnamic Acid Alleviates Rheumatoid Arthritis by Inhibiting TLR4/NFκB/NLRP3 Activation-Induced Pyroptosis

Abstract

Growing evidence shows that Baihu-Guizhi decoction (BHGZD), a traditional Chinese medicine (TCM)-originated disease-modifying anti-rheumatic prescription, may exert a satisfying clinical efficacy for rheumatoid arthritis (RA) therapy. In our previous studies, we verified its immunomodulatory and anti-inflammatory activities. However, bioactive compounds (BACs) of BHGZD and the underlying mechanisms remain unclear. Herein, an integrative research strategy combining UFLC-Q-TOF-MS/MS, gene expression profiling, network calculation, pharmacokinetic profiling, surface plasmon resonance, microscale thermophoresis, and pharmacological experiments was carried out to identify the putative targets of BHGZD and underlying BACs. After that, both in vitro and in vivo experiments were performed to determine the drug effects and pharmacological mechanisms. As a result, the calculation and functional modularization based on the interaction network of the “RA-related gene–BHGZD effective gene” screened the TLR4/PI3K/AKT/NFκB/NLRP3 signaling-mediated pyroptosis to be one of the candidate effective targets of BHGZD for reversing the imbalance network of “immune-inflammation” during RA progression. In addition, both mangiferin (MG) and cinnamic acid (CA) were identified as representative BACs acting on that target, for the strong binding affinities between compounds and target proteins, good pharmacokinetic features, and similar pharmacological effects to BHGZD. Notably, both BHGZD and the two-BAC combination of MG and CA effectively alleviated the disease severity of the adjuvant-induced arthritis-modified rat model, including elevating pain thresholds, relieving joint inflammation and bone erosion via inhibiting NF-κB via TLR4/PI3K/AKT signaling to suppress the activation of the NLRP3 inflammasome, leading to the downregulation of downstream caspase-1, the reduced release of IL-1β and IL-18, and the modulation of GSDMD-mediated pyroptosis. Consistent data were obtained based on the in vitro pyroptosis cellular models of RAW264.7 and MH7A cells induced by LPS/ATP. In conclusion, these findings offer an evidence that the MG and CA combination identified from BHGZD may interact with TLR4/PI3K/AKT/NFκB signaling to inhibit NLRP3 inflammasome activation and modulate pyroptosis, which provides the novel representative BACs and pharmacological mechanisms of BHGZD against active RA. Our data may shed new light on the mechanisms of the TCM formulas and promote the modernization development of TCM and drug discovery.

Targeting pyroptosis as a preventive and therapeutic approach for stroke

“… Regulation of M1/M2 microglia, regulation of autophagy, and anti-apoptosis PGC-1α; Sirt1; JAK2/STAT3/PI3K/AKT/mTOR MCRO/R; OGD [ 93 , 113 , 114 ] Mangiferin Anemarrhena asphodeloides Bge. Anti-inflammatory, antioxidant, and anti-apoptotic TLR4/NLRP3; SIRT1/PGC-1α MCAO/R; OGD/R [ 115 , 116 ] Salidroside Rhodiola rosea L. Anti-inflammatory, antioxidant, and anti-apoptotic TLR4/NLRP3; Bax/Bcl-2; Nrf2/Trx1 MCAO/R; OGD/R [ 117 , 118 ] Schisandrin B Schisandra chinensis (Turcz.) Baill.…”

Anti-Inflammation and Anti-Pyroptosis Activities of Mangiferin via Suppressing NF-κB/NLRP3/GSDMD Signaling Cascades

“…For example, mangiferin significantly reduced NO, IL-1β, IL-6, and TNF-α by inhibiting activation of NF-κB in LPS-induced mouse microglia cells [ 18 ], and decreased IL-1β, IL-6, and TNF-α by inhibiting the nuclear translocation of the NF-κB p65 subunit in mononuclear macrophages, and protected mouse lung injury against sepsis [ 19 ]. Meanwhile, mangiferin may interact with TLR4/PI3K/AKT/NFκB signaling to inhibit NLRP3 inflammasome activation and modulate GSDMD-mediated pyroptosis in rats [ 20 ], and it alleviated the histopathology of inflammation and the levels of pro-inflammatory cytokines (TNF-α, IL-1β, and IL-6) via inhibiting NF-κB and NLRP3 inflammasome activation in lactating female mice [ 21 ]. Therefore, mangiferin has significant pharmacological function in inhibiting inflammation and inhibiting pyroptosis in both cell models and mammalian models.…”

Pyroptosis by NLRP3/caspase‐1/gasdermin‐D pathway in synovial tissues of rheumatoid arthritis patients

“…Our experiments support the involvement of pyroptosis mediated by the NLRP3 inflammasome, caspase-1 and GSDMD 29 in RA, providing clinical support for similar results obtained with rat models of RA. 28,30 Our results may help explain the therapeutic effects of tofacitinib, a clinically licensed drug that works against the disease by inhibiting activation of the NLRP3 inflammasome. 31 Other pyroptosis pathways may also contribute to RA.…”