COX-2 x HSP90

Microglia function as a vital constituent in the maintenance of brain homeostasis. Aberrant microglial activation, however, may contribute to neurodegenerative diseases. Lithospermic acid (LA) is a plant-derived polycyclic phenolic carboxylic acid isolated from Salvia miltiorrhiza. The present study investigated the potential effects of lithospermic acid on LPS-induced neuroinflammation in BV2 microglial cells and determined the mechanism of action of this compound. Cells were pre-treated with lithospermic acid for 1 h and incubated with LPS for 24 h. qPCR, immunofluorescence, and immunoblot assays were used to determine the expression of iNOS, COX2, NF-κB p65, and HSP90 expression. ELISA was employed to measure the production of pro-inflammatory cytokines. Lithospermic acid dramatically reduced LPS-stimulated cell migration and decreased NF-κB p65 nuclear translocation. Furthermore, lithospermic acid also markedly decreased the production of pro-inflammatory cytokines, including IL-6, IL-1β, and TNF-α in a dose-dependent manner. Additionally, lithospermic acid inhibited NO and PGE2 production in response to LPS, and it also inhibited the expression of iNOS and COX2 in a dose-dependent manner. Molecular docking and experimental verification have demonstrated that lithospermic acid inhibits the activity and expression of HSP90. Small interfering RNA knockdown of HSP90 expression, which abrogated LPS-induced inflammation. These findings suggest that the lithospermic acid targeting HSP90 attenuates LPS-induced inflammatory response via the NF-κB signalling pathway in BV2 microglial cells. Collectively, lithospermic acid may offer therapeutic benefits for neurodegenerative disorders associated with microglial activation and could serve as a potential inhibitor/agent for the treatment of neuroinflammation.

Diclofenac, a widely used nonsteroidal anti-inflammatory drug (NSAID), is effective in treating pain and inflammation and has also shown potential as an anticancer agent, primarily through cyclooxygenase-2 (COX-2) inhibition. However, its therapeutic application is often limited by adverse effects, necessitating the development of new derivatives with improved pharmacological profiles. In this study, two novel diclofenac hydrazide-imine derivatives, DDCH (cyclohexanone-based) and DDAC (acetylacetone-based), were synthesized, structurally characterized, and evaluated through combined computational and experimental approaches. Density functional theory (DFT) calculations provided transition-state and energy profile analyses, while molecular docking and molecular dynamics (MD) simulations established stable interactions of both derivatives with COX-2 and heat shock protein 90 (HSP90), a key oncogenic chaperone. DDAC exhibited particularly strong binding to HSP90, suggesting enhanced anticancer potential compared with diclofenac. In silico drug-likeness and ADME assessments, including Lipinski's Rule of Five, predicted favorable pharmacokinetic properties. Experimental evaluation confirmed anti-inflammatory efficacy: in vitro protein denaturation assays showed that DDAC display inhibition comparable to diclofenac, while in vivo carrageenan-induced paw edema studies demonstrated significant activity for both derivatives. Collectively, these findings confirm that DDCH and DDAC retain anti-inflammatory properties while offering predicted anticancer potential, with DDAC emerging as the more promising dual-action candidate. This work establishes a rational framework for the further optimization and pharmacological development of diclofenac-based analogues targeting both inflammation and cancer.

Diabetic nephropathy (DN) is the leading cause of end-stage renal disease (ESRD) worldwide, underscoring the urgent need to explore effective strategies for its prevention and control. Accumulating evidence indicates that inhibition of abnormal mesangial cells (MCs) proliferation, particularly during the early stages of DN, represents a critical therapeutic strategy for attenuating the progression of DN. Complanatoside A (CA), a major flavonoid derived from Semen Astragali Complanati, has demonstrated significant reno-protective properties. In this study, diabetic mouse models and high-glucose (HG)-induced mouse mesangial cell models were established to investigate the reno-protective mechanisms of CA. Furthermore, network pharmacology was employed to predict molecular targets and signaling pathways. Histopathological analysis revealed that CA alleviated key renal pathological changes, including glomerular interstitial fibrosis, thickening of the glomerular basement membrane, mesangial matrix expansion, glomerulosclerosis, and fibrillar collagen deposition. Furthermore, CA inhibited HG-induced mesangial extracellular matrix (ECM) accumulation, inflammatory responses, and cellular proliferation. Through network pharmacology, eight core genes (TNF-α, AKT1, HSP90AA1, MMP9, PPARG, SRC, PTGS2, and MMP2) were identified as critical targets of CA in DN and were primarily associated with inflammatory responses and ECM deposition. Molecular docking analysis confirmed that CA exhibited high binding affinity for these inflammation- and ECM-related genes. This study demonstrated that CA effectively mitigated renal injury in DN by suppressing inflammatory pathways and ECM deposition, thus providing novel insights into its therapeutic potential. These findings offer new perspectives for the development of traditional Chinese medicine-based interventions for the treatment of DN.