Activation of adenosine A3 receptor attenuates progression of osteoarthritis through inhibiting the NLRP3/caspase‐1/GSDMD induced signalling

Abstract

The specific adenosine A3 receptor (A3AR) agonist (CF101) has potential for inflammation and pain in various disease, such as arthritis, cancer and neuropathic pain, while the role of A3AR in post‐traumatic OA and the underlying mechanism is largely unknown. CF101 was orally administrated in OA rats induced by anterior cruciate ligament transection (ACLT) surgery, and the rat primary chondrocytes were stimulated by hydrogen peroxide (H2O2, 300 μM). Histologic grading system was performed for detecting cartilage degeneration and immunohistochemistry for determining pyroptosis. The moleculars associated with cartilage homeostasis and inflammatory cytokines were analysed; moreover, the activation of NLRP3 inflammasome was determined. CF101 treatment significantly attenuated OA cartilage damage, OA‐related pain and cartilage pyroptosis. Chondrocytes stimulated by H2O2 evoked ROS release, thereby promoting the activation of NLRP3 inflammasome and facilitating the cleavage of GSDMD, which ultimately resulted in the mass release of pro‐inflammatory cytokines including IL‐1β and IL‐18, and production of matrix hydrolase. The pre‐treatment with CF101 powerfully inhibited the above process both in vivo and in vitro. Our findings demonstrated that activation of A3AR attenuates OA progression and relieves pain perception through suppression of cartilage degradation and inhibition of ROS/NLRP3/GSDMD signalling, indicating pyroptosis is a potential candidate for OA treatment.

Recent Therapeutic Strategies for Excessive Chondrocyte Death in Osteoarthritis: A Review

“…Mesenchymal stem cell‐derived exosomes, microRNAs, phytochemical, growth factors, biological enzymes, and exercise all exhibit good potential to inhibit chondrocyte pyroptosis. In addition, inhibitors of pyroptosis‐related pathways have also been shown to alleviate OA by reducing pyroptosis, as seen in Table 4 173–186 …”

Pyroptosis in inflammatory bone diseases: Molecular insights and targeting strategies

“…Nrf2/HO-1 axis inhibits NF-κB pathway to attenuate pyroptosis 88 SDF-1 activates AMPK signal to suppress NLRP3 inflammasome and subsequent pyroptosis 89 A3AR inhibits the ROS/NLPR3/GSDMD axis to relieve pyroptosis-mediated symptoms in OA 90 Osteomyelitis NLRP3 inflammasome is activated to cause bone loss 93 Periodontitis Hypoxia with P. gingivalis-LPS synergistically activate NLRP3 to induce pyroptosis 101 MARK4 induces NLRP3-mediated pyroptosis 102 The Nrf2/HO-1 axis inhibits LPS-induced ROS to prevent NLRP3-mediated pyroptosis 98 NLRP6 inflammasome activates caspase-1 to induce pyroptosis of HGFs 103 Caspase-4/GSDMD pathway mediates noncanonical pyroptosis 104,105 Butyrate activates caspase-3/GSDME axis to induce pyroptosis 106 LPS-induced Dec1 upregulates NF-κB via the PI3K/AKT pathway to promote pyroptosis, while LPS-induced Dec2 attenuates the phosphorylation of NF-κB to inhibit pyroptosis [108][109][110][111] RA TNF-α plus CRT induces NLRP3 inflammasome-mediated pyroptosis 116 ACPAs induces NLRP3 inflammasome-mediated pyroptosis via the Akt/NF-κB axis 117 S100A9 may participate in the IL-6-induced pyroptosis via activating the CTSB/ATP pathway 118 Hypoxia induces NLRP3 inflammasome-mediated pyroptosis via the ROS/GRK2/HIF-1α axis 119 TNF-α plus hypoxia induce caspase-3/GSDME-mediated pyroptosis 120 ASIC1a promotes the influx of Ca 2+ , activating the calpain/calcineurin axis to induce NLRP3 inflammasome-mediated pyroptosis 122,123 CaSR promotes the influx of Ca 2 to induce NLRP3 inflammasome-mediated pyroptosis 124 Abbreviations: ACPAs, anti-citrullinated protein antibodies; AP, apical periodontitis; ASIC1a, acid-sensitive ion channel 1a; ATP, adenosine triphosphate; A3AR, adenosine A3 receptor; CaSR, calcium-sensing receptor; CRT, calreticulin; CTSB, cathepsin B; Dec1, embryo-chondrocyte expressed genes 1; FLS, fibroblast-like synoviocyte; Gout, gouty arthritis; GRK2, G protein-coupled receptor kinase 2; GSDMD, gasdermin D; HGFs, human gingival fibroblasts; HIF-1α, hypoxia-inducible factor-1α; HO-1, heme oxygenase-1; IL, interleukin; LPS, lipopolysaccharide; MARK4, microtubule affinity regulating kinase 4; NF-κB, nuclear factor kappa-B; NLRP3, nucleotide-binding oligomerization domain receptor family PYD domain containing 3; NOX, NAD(P)H oxidases; Nrf2, nuclear factor erythroid-2-related factor 2; OA, osteoarthritis; P. gingivalis, Porphyromonas gingivalis; ROS, reactive oxygen species; SDF-1, stromal cell-derived factor-1; TNF-α, tumor necrosis factor-α; USP7, ubiquitin-specific protease 7.…”

“…Also, the specific adenosine A3 receptor (A3AR) also slows the course of OA and eases pain by inhibiting the axis of ROS/NLRP3/GSDMD. 90…”

“…Direct inhibitors of the NLRP3 inflammasome include CY-09, 128 glyburide, 93 icariin, 82 MCC950, 21,98,99 monomeric derivatives of paeoniflorin (MDP), 119 metformin, 129 cranberry proanthocyanidins (PACs), 130 and resveratrol. 75 Indirect NLRP3 inhibitors include A3AR activator (CF101), 90 ASIC-1a inhibitors (amiloride, psalmotoxin-1), 122 GRK2 phosphorylation inhibitor (MDP), 119 glycogen synthase kinase-3β (GSK-3β) inhibitor (GSKI), 131 NF-κB inhibitors (Baeckein E, loganin, morroniside, PDTC, punicalagin), 21,[132][133][134][135] Nrf2/HO-1 activators (eldecalcitol, licochalcone A), 88,98 and ROS inhibitors (enamel matrix derivative, NAC, quercetin). 98,99,136,137 5.2 | MicroRNAs (miRNAs) targeting strategies Epigenetic processes, including DNA methylation, histone modifications, and miRNAs, which causing the heritable silence of genes without alteration to their coding sequence.…”

“…In addition, Wang et al 89 demonstrated that stromal cell‐derived factor‐1 (SDF‐1), a homeostatic CXC chemokine, could activate the AMPK signaling pathway to suppress NLRP3 inflammasome and subsequent pyroptosis, alleviating the symptoms of OA. Also, the specific adenosine A3 receptor (A3AR) also slows the course of OA and eases pain by inhibiting the axis of ROS/NLRP3/GSDMD 90 …”