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Aim:To investigate the effect of cholecystokinin octapeptide(CCK-8)on thediacylglycerol-protein kinase C(DAG-PKC)signaling pathway in rat pulmonaryinterstitial macrophages(PIM)stimulated by lipopolysaccaride(LPS).Methods:The PIM from rat lung tissues were isolated using the collagenase digestionmethod combined with alveolar lavage and pulmonary vessel perfusion.DAGcontent and PKC activity were measured by radioenzymatic assay.The transloca-tion of PKCζ was determined by semi-quantitative immunoblot analysis.Results:CCK-8,at high concentrations(1×10~(-6)-1×10~(-5)mol/L),decreased DAG content andinhibited PKC activity and PKCζ translocation compared with that in rat restingPIM of a control group(P<0.01).LPS increased DAG content,and promoted PKCactivity and PKC~ translocation(P<0.01).CCK-8 decreased LPS-induced DAGcontent and inhibited LPS-induced PKC activity and PKCζ translocation signifi-cantly at 1×10~(-8)-1×10~(-5)mol/L(P<0.01).This inhibitory effect of CCK-8 could beabrogated partly by proglumide(non-selective CCK receptor antagonist),CR-1409(selective CCK-A receptor antagonist),and CR-2945(selective CCK-Breceptor antagonist)in a concentration-dependent manner(P<0.01).Conclusion:CCK-8 was a negative modulator of the DAG-PKC signaling pathway in rat restingPIM,which is very important for maintaining body homeostasis.It significantlyinhibited LPS-induced DAG content,PKC activity and PKCζ translocation in aconcentration-dependent manner.The CCK receptor,especially the CCK-Areceptor,might play a major role in this process.
Aim: To investigate the effect of cholecystokinin octapeptide (CCK-8) on the diacylglycerol-protein kinase C (DAG-PKC) signaling pathway in rat pulmonary intiti macrophages (PIM) stimulated by lipopolysaccaride (LPS). Methods: The PIM from rat lung isolated using the collagenase digestionmethod combined with alveolar lavage and pulmonary vessel perfusion. DAGcontent and PKC activity were measured by radioenzymatic assay. transloca tion of PKCζ was determined by semi-quantitative immunoblot analysis. Results: CCK-8, at high concentrations (1 × 10-6 (-6) -1 × 10-5 mol / L), decreased DAG content and inhibitor PKC activity and PKCζ translocation compared with that in rat resting PIM control group (P <0.01) .LPS increased DAG content , and promoted PKC activity and PKC ~ translocation (P <0.01) .CCK-8 decreased LPS-induced DAGcontent and inhibited LPS-induced PKC activity and PKCζ translocation signifi-cantly at 1 × 10 -8 -1 × 10 ~ -5) mol / L (P <0.01). This inhibitory effect of CCK-8 could be beabrogate CR-1409 (selective CCK-A receptor antagonist), and CR-2945 (selective CCK-Breceptor antagonist) in a concentration- dependent manner (P <0.01) .Conclusion: CCK-8 was a negative modulator of the DAG-PKC signaling pathway in rat resting PIM, which is very important for maintaining body homeostasis. Significantly inhibited LPS-induced DAG content, PKC activity and PKCζ translocation in aconcentration-dependent manner. CCK receptor, especially the CCK-Areceptor, might play a major role in this process.