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Background: Pyroptosis is involved in the occurrence of acute pancreatitis, but its role in remote organ injury remains unclear. Aims: To investigate the role and mechanism of NLRP3 inflammasome-dependent pyroptosis in acute pancreatitis- related lung injury. Methods: Thirty-two male Sprague-Dawley rats were randomly divided into four groups: control group, severe acute pancreatitis (SAP) group, Z-WEHD-FMK (caspase-1 inhibitor) group and disulfiram (GSDMD inhibitor) group. Experimental SAP was constructed by using 5% sodium taurocholate in the latter 3 groups. Serum levels of amylase, lipase, procalcitonin, and the myeloperoxidase (MPO) activity were determined; the severity of pancreatic and lung injuries was assessed by histopathology and lung wet/dry weight ratio; serum levels of pyroptosis-related inflammatory cytokines and the expressions of proteins involved in pyroptosis pathway in lung tissue were measured by ELISA method and immunohisto- chemistry and Western blotting, respectively. Results: Compared with the control group, the serum biochemical indices, MPO activity, and interleukin (IL)-1β, IL-18 levels in SAP group were significantly increased with aggravated pancreatic and lung tissue injuries; meanwhile, the expressions of NLRP3, caspase-1 and GSDMD in lung tissue were significantly up- regulated (all P<0.05). Pretreatment with caspase-1 or GSDMD inhibitors reduced the severity of pancreatic and lung tissue injuries, improved the serum biochemical indices and MPO activity, and ameliorated the increased pyroptosis - related inflammatory cytokines and pyroptosis pathway - related proteins (all P<0.05). Conclusions: NLRP3/caspase - 1/GSDMD pathway mediated pyroptosis plays an important role in acute pancreatitis-related lung injury, and inhibition of pyroptosis pathways might be a new direction for its treatment.
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Background:TLR4/NF-κBp65 signaling pathway plays an important role in triggering inflammatory response,and regulates releasing of cytokines in acute pancreatitis. However,the role of this pathway in inflammation in severe acute pancreatitis(SAP)associated with acute kidney injury(AKI)is not clear. Aims:To investigate the effect of TLR4/NF-κBp65 signaling pathway on AKI in experimental SAP. Methods:Thirty-two male Sprague-Dawley rats were randomly assigned into 4 groups(8 each):normal control group,SAP 6 h,12 h,and 18 h groups. SAP was induced by retrograde injection of 4% sodium taurocholate into biliopancreatic duct. Serum levels of creatinine(Cr)and blood urea nitrogen (BUN)were measured dynamically. Pathological changes of kidney were observed macro- and microscopically. Serum levels of tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)were determined by ELISA,and the localization and expressions of TLR4 and NF-κBp65 in kidney were determined by immunohistochemical staining and Western blotting. Results:Compared with normal control group,the kidney injuries in SAP groups were gradually aggravated with disease progression;meanwhile,serum levels of Cr,BUN,TNF-α and IL-6 increased significantly,and the expressions of TLR4 and NF-κBp65 in kidney became more intensive(P all <0.05). Expressions of TLR4 and NF-κBp65 in kidney were positively correlated with the serum levels of Cr,BUN,TNF-α and IL-6. Conclusions:In experimental SAP,the changes of TLR4 and NF-κBp65 expressions in kidney are coincidence with the severity of kidney injury and the serum levels of proinflammatory cytokines,which indicates that TLR4/NF-κBp65 signaling pathway plays an important proinflammatory effect in disease progression of SAP associated with AKI.
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Objective The mechanism of celecoxib in influencing the migration of vascular endothelial cells is still not clear.The aim of this study was to investigate the effect of celecoxib against migration of vascular endothelial cells and its mechanism.Methods Human umbilical vein endothelial cells (HUVECs) were obtained from the cell bank of Central Laboratory of Nanjing General Hospital of Nanjing Military Command.Transwell was used to measure the migration rate of HUVECs after the administration of 0μmol/L, 10μmol/L and 20μmol/L celecoxib.Immunofluorescence, western blot, and qRT-PCR were used to detect protein and mRNA expression of COX-2 and PTPRJ in HUVECs.Results The expression of COX-2, PTPRJ in HUVECs were detected by immunofluorescence.After the administration of 20μmol/L, 10μmol/L and 0μmol/L celecoxib, HUVEC migration rate was (12.35±3.61), (32.80±5.92) and (63.15±5.83) respectively, representing significant difference (P<0.01).COX-2 protein expression in 20μmol/L group (0.16±0.03) and 10μmol/L group (0.36±0.05) decreased significantly compared with that of 0μmol/L group (0.77±0.07) (P<0.01).Moreover, COX-2 protein expression in 20μmol/L group significantly decreased compared with that of 10μmol/L group(P<0.05).PTPRJ protein expression in 20μmol/L group (0.82±0.05) and 10μmol/L group (0.51±0.02) was respectively higher than that of 0μmol/L group (0.27±0.04) (P<0.01).Moreover, PTPRJ protein expression in 20μmol/L group significantly increased compared with that of 10μmol/L group (P<0.05).COX-2 mRNA expression in 20μmol/L group (0.06±0.02) and 10μmol/L group (0.22±0.05) decreased significantly compared with that of 0μmol/L group (1.05±0.13) (P<0.01).PTPRJ mRNA expression in 20μmol/L group (60.27±11.31) and 10μmol/L group (16.50±3.18) increased significantly compared with that of 0μmol/L group (0.99±0.25) (P<0.01).Conclusion Celecoxib inhibits the migration of vascular endothelial cells, which may be related to the inhibition of COX-2 expression and the up-regulation of PTPRJ expression in vascular endothelial cells.