Astilbin inhibits pancreatic stellate cell activation via the NLRP3 inflammasome
CUI Li-hua,LI Di-hua,ZHANG Lan-qiu
Abstract:
Objective To investigate the mechanism by which astilbin (AST) regulates the activation of pancreatic stellate cells (PSCs) via the NOD-like receptor family, pyrin domain containing 3 (NLRP3) inflammasome. Methods PSCs were isolated from normal pancreatic tissue and cultured in vitro. After treatment with different concentrations of AST for 24 h and 48 h, the effect of AST on PSC viability was detected using the CCK-8 assay to determine the appropriate concentration for in vitro experiments. PSCs were treated with the selected concentration of AST for 24 h. The expression of PSC activation markers:α-smooth muscle actin (α-SMA), collagen type I (Collagen-I), and fibronectin (FN)，as well as NLRP3 inflammasome related markers (NLRP3, Caspase-1, and IL-18) was examined by real-time quantitative PCR (RT-qPCR), Western blot, and immunofluorescence staining. To further confirm the key role of NLRP3 in AST mediated regulation of PSC activation, PSCs were pre-treated with the NLRP3 promoter LPS or the NLRP3 inhibitor MCC950 for 1 h, followed by AST co-incubation for 24 h. Results PSC viability was significantly affected with increasing AST concentrations. Concentrations of 100, 200, and 500 μg/mL AST (24 h treatment) were selected for subsequent experiments. Compared with the control group (0 μg/mL), different concentrations of AST significantly reduced the mRNA and protein expression of FN, Collagen-I, α-SMA, NLRP3, Caspase-1, and IL-18 (P <0.05). At 500 μg/mL, AST inhibited the LPS-induced increase in NLRP3 and Collagen-I expression (P <0.05). Moreover, the regulatory effects of AST on NLRP3 and Collagen-I were suppressed by MCC950. Conclusion AST inhibits PSC activation and reduces extracellular matrix deposition by targeting and suppressing NLRP3 activation, thereby providing a new theoretical basis for the treatment of pancreatic fibrosis.
