In vitro expression of CD14 protein and its gene in Kupffer cells induced by lipopolysaccharide

摘要

OBJECTIVES: To observe expression of CD14 protein and its gene in Kupffer cells induced bylipopolysaccharide (LPS) and explore the role of CD14 in LPS-induced Kupffer cell activation.METHODS: Kupffer cells were isolated from Wistar rat livers by in situ collagenase digestion, followedby culture and incubation with 100μg/ml LPS for 0, 30, 60 and 120 min, respectively. CD14 proteinexpressed on the membrane of Kupffer cells was examined using confocal microscopy and Western blottinganalysis. Expression of CD14 mRNA in Kupffer cells was determined by reverse transcription polymerasechain reaction (RT-PCR). Tumor necrosis factor (TNF)-α and interleukin (IL)-6 in the supernatantwere measured using enzyme-linked immunosorbent assay (ELISA) kit.RESULTS: Expression of CD14 mRNA and CD14 protein in isolated Kupffer cells was upregullatedquickly after LPS stimulation and increased with time. Likewise, there was a time-dependent increase ofTNF-α and IL-6 in the supernatant with upregulation of CD14 expression. There were significantdifferences between normal and LPS-stimulated Kupffer cells (P0. 05).CONCLUSIONS: LPS can upregulate expression of CD14 protein and its gene in isolated Kupffer cells.CD14 may play an important role in the activation of LPS-induced Kupffer cells.