Expression Of Protein Kinase C And Cytokine In Airway Epithelial Cell Interfered By Lipopolysaccharide

OBJECTIVES:Lipopolysaccharide stimulated the airway epithelial cells secreting a variety ofcytokines. in the present study，related signaling pathways mainly included p38mitogen-activated protein kinase、c-Jun N-terminal protein kinase and extracellularsignal-regulated kinase1/2. Protein kinase C was also a common signaling pathways,studies had shown that PKC was not only involved in the process of type II alveolarepithelial cells secreting cytokines, but also affected the blood-brain barrier andintestinal epithelial barrier function. There was little study about the role of proteinkinase C in the airway epithelial cells. This experiment is mainly to detectconcentration of different cytokines (IL-1、IL-6、IL-8、IL-11) and expression of PKCafter LPS stimulation airway epithelial cells at the same time. To explore the role ofPKC in the airway epithelial cells secreting a variety of cytokines.METHODS:A549cell strain was cultivated and interfered by different concentrations LPS，divided into control group (0μg/ml) and experimental groups(0.1μg/ml、1μg/ml、10μg/ml、20μg/ml、30μg/ml、50μg/ml、100μg/ml)，after2h、4h、8h、24h、30hand48h，We collected culture medium and total protein in the cell，detected volumeof cytokines IL-1、IL-6、IL-8and IL-11secreted by A549cell using enzyme-linkedimmunosorbent assays (ELISA).Meanwhile, we measured expression of PKC bywestern blotting.RESULTS:1.There were significantly differences in volume of cytokines IL-1、IL-6、IL-8、IL-11and expression of PKC interfered by various concentration of LPS in sameperiod（all P values＜0.001）；Compared with the control group, the maximumsecretion of IL-1and the maximum expression of PKC were increased at all the time，the difference was statistically significant (P value<0.001)，the maximum secretionof IL-6(2h、4h、24h、48h)、IL-8（2h、4h、8h、24h、30h）and IL-11（2h、8h、 24h、30h、48h）were increased at some time，the difference was statistically significant(P value<0.001), whereas the secretion of IL-6was decreased at8h and30h.2.Expression of PKC was correlated with volume of cytokine IL-11(R=0.127),the difference was statistically significant (P=0.049).CONCLUSIONS:1. Secretion of cytokine IL-1、IL-6、IL-8and IL-11and expression of PKC areinterfered by LPS；LPS increased the secretion of IL-1（2h、4h、8h、24h、30h、48h）、IL-8（2h、4h、8h、24h、30h）、IL-11（2h、8h、24h、30h、48h）and theexpression of PKC in airway epithelial cells at certain times; whereas the secretion IL-6was not correlated with the concentration of LPS.2. Expression of PKC was positive correlated with volume of cytokine IL-11sμggested that secretion of IL-11interfered by LPS may be correlation with PKCsignal pathway.