摘要
AIM:To observe synthesis of CD14 protein and expressionof CD14 mRNA in hepatic tissue and hepatocytes of ratsduring endotoxemia.METHODS:The endotoxemia model of Wistar rat wasestablished by injection of a dose of lipopolysaccharide(LPS)(5mg·kg^(-1),Escherichia coil O111:B4)via the tailvein,then the rats were sacrificed after 3,6,12 and 24 h inbtaches.Hepatocytes were isolated from normal and LPS-injected rats by in situ collagenase perfusion technique andwere collected to measure the expression of CD14 mRNAand synthesis of CD14 protein by reverse transcription-polymerase chain reaction(RT-PCR)or Western blotanalysis.The binding of fluorescein isothiecyanate(FITC)-CD14 polyclonal antibody to isolated hepatocytes was alsoassessed by flow cytometric analysis(FCM).RESULTS:In the rats with endotoxemia,the expressions ofCD14 mRNA in hepatic tissue and isolated hepatocytes werestronger at 3,6,and 12 h than that in control rats(3.48±0.15,5.89±0.62,4.33±0.18,vs 1.35±0.14 in hepatictissue,P<0.01;4.12±0.17,6.24±0.64,4.35±0.18,vs1.87±0.15 in hepatecytss,P<0.01).The synthesis of CD14protein in hepatic tissue and isolated hepatoeytes increasesalso obviously in 6 and 12 h when compared to that incontrol rats(13.27±1.27,17.32±1.35,11.42±1.20,vs 7.34±0.72 in hepatic tissue,P<0.01;14.68±_+1.30,17.95±1.34,11.65±1.19,vs 7.91±0.70 in hepatocytoes,P<0.01).FCM showed that mean fluorescence intensity(MFI)andnumbers of FITC-CD14 positive cells in the rats withendotoxemia increased obviously at 3,6,12 and 24h whencompared with normal control group(43.4%,70.2%,91.4%,32.6% vs4.5%,P<0.01).CONCLUSION:LPS can markedly promote the synthesis ofCD]4 protein and up-regulate the expression of CD14 mRNAin isolated hepatocytes and hepatic tissue.Liver might be amain source for soluble CD14 production duringendotoxemia.
AIM:To observe synthesis of CD14 protein and expression of CD14 mRNA in hepatic tissue and hepatocytes of rats during endotoxemia. METHODS:The endotoxemia model of Wistar rat was established by injection of a dose of lipopolysaccharide (LPS)(5mg·kg^(-1),Escherichia coil O111:B4)via the tail vein,then the rats were sacrificed after 3,6,12 and 24 h in btaches.Hepatocytes were isolated from normal and LPS- injected rats by in situ collagenase perfusion technique and were collected to measure the expression of CD14 mRNA and synthesis of CD14 protein by reverse transcription- polymerase chain reaction(RT-PCR)or Western blot analysis.The binding of fluorescein isothiecyanate(FITC)- CD14 polyclonal antibody to isolated hepatocytes was also assessed by flow cytometric analysis(FCM). RESULTS:In the rats with endotoxemia,the expressions of CD14 mRNA in hepatic tissue and isolated hepatocytes were stronger at 3,6,and 12 h than that in control rats(3.48± 0.15,5.89±0.62,4.33±0.18,vs 1.35±0.14 in hepatic tissue,P<0.01;4.12±0.17,6.24±0.64,4.35±0.18,vs 1.87±0.15 in hepatecytss,P<0.01).The synthesis of CD14 protein in hepatic tissue and isolated hepatoeytes increases also obviously in 6 and 12 h when compared to that in control rats(13.27±1.27,17.32±1.35,11.42±1.20,vs 7.34 ±0.72 in hepatic tissue,P<0.01;14.68±_+1.30,17.95±1.34, 11.65±1.19,vs 7.91±0.70 in hepatocytoes,P<0.01). FCM showed that mean fluorescence intensity(MFI)and numbers of FITC-CD14 positive cells in the rats with endotoxemia increased obviously at 3,6,12 and 24h when compared with normal control group(43.4%,70.2%, 91.4%,32.6% vs4.5%,P<0.01). CONCLUSION:LPS can markedly promote the synthesis of CD]4 protein and up-regulate the expression of CD14 mRNA in isolated hepatocytes and hepatic tissue.Liver might be a main source for soluble CD14 production during endotoxemia.
基金
the National Natural Science Foundation of China(No.39970719)
