Fig 1: Further validation of the mechanistic pathway of Hyperibone J (HJ) action. (A) The knockdown rate of ADK mRNA level (n = 3). (B-H) Expression of IL-1β, IL-6, TNF-α, TLR4 in LPS-stimulated BV-2 cell with ADK knockdown (n = 3). (I) Effects of HJ on the expressions of NLRP3, P2X7R, p-P65 (p-Ser536), total-P65, pro-IL-1β, maturate-IL-1β, pro-Caspase-1 and cleaved-Caspase-1 protein in LPS-stimulated BV-2 cell (n = 3). (J-K) Effects of HJ on the production and release of ATP in LPS-stimulated BV-2 cell (n = 3). (L) The knockdown rate of ADK protein level (n = 3). (M) Protein expression of NLRP3 and mature-IL-1β in LPS-stimulated BV-2 cell with ADK knockdown (n = 3). (N) After the treatment with A438079, changes in effects of HJ on the production of cleaved-Caspase-1 and mature IL-1β in LPS-stimulated BV-2 cell (n = 3). *P < 0.05, **P < 0.01, ***P < 0.005, ****P < 0.001 vs. LPS.
Fig 2: Mechanisms of Hyperibone J exerts antidepressant effects. Hyperibone J could target ADK, reducing the upregulation of ADK expression induced by LPS, thereby decreasing ATP production from phosphorylated adenosine and inhibiting the ATP/P2X7R signaling pathway leading to Caspase-1/IL-1β cascade cleavage. Meanwhile, ADK could regulate the transcription of Tlr4, thereby affecting the TLR4/NF-κB pathway and reducing the transcription of Il-1b, Tnf, and Il-6, thus exerting antidepressant activity by inhibiting neuroinflammation.
Supplier Page from Abcam for Recombinant human ADK protein (Active) (His tag N-Terminus)