Objective: To observe the expression of CD14 mRNA and its protein in rat hepatocytes during endotoxemia.
Methods: The acute endotoxemia model of wistar rats was established by injection of a dose of LPS (5mg/kg, Escherichia coli O111:B4) via the tail vein, then the rats were sacrificed at 3, 6, 12, and 24 hour, respectively. Hepatocytes were isolated from normal and LPS-injected rats by in situ collagenase perfusion technique. Hepatocytes and hepatic tissue were collected to measure the expression of CD14 mRNA and CD14 protein by reverse transcriptase-polymerase chain reaction (RT-PCR) or Western blot analysis. The binding of fluorescein isothiocyanate (FITC)-CD14 polyclone antibody to isolated hepatocytes was also assessed by flow cytometric analysis (FCM).
Results: In the rats with endotoxemia, the expression of CD14 mRNA and CD14 protein in hepatic tissue and isolated hepatocytes were stronger at 6, 12 and 24 hour than that in control rats (P<0.01). FCM showed that positive cell numbers of FITC-CD14 in the rats with endotoxemia increased when compared with normal control group (P<0.01).
Conclusions: LPS can markedly upregulate the expression of CD14 mRNA and CD14 proteins in hepatic tissue and isolated hepatocytes.