Association of CD14 and TLR4 with LPS-stimulated human normal skin fibroblasts in immunophenotype changes and secretion of TGF-beta 1 and IFN-gamma

作者：Yang, Hongming^*; Li, Juncong; Wang, Yihe; Hu, Quan

来源：International Journal of Clinical and Experimental Pathology, 2015, 8(2): 1991-1995.

• Lipopolysaccharide
• fibroblast
• collagen
• toll like receptor 4
• CD14

Objectives: We attempted to explore the association of CD14 and TLR4 with LPS-stimulated human normal skin fibroblasts in immunophenotype changes and secretion of TGF-beta 1 and IFN-gamma, and to expand the current knowledge of the mechanisms that underlie LPS-induced scar formation. Methods: We randomized the human normal skin fibroblasts cultured in vitro into four groups. The expression profile of immune phenotypes was determined by immunohistochemical staining. Ultrastructure of cells was observed by use of transmission electron microscopy. Secretion status of TGF-beta 1 and IFN-gamma was inspected using ELISA assay. Results: Compared with group A, the expressions of alpha-SMA and alpha 1 (I) procollagen in groups B, C, D were lower, and it in group D were the lowest in all groups. The cells in group A were diversification under the electron microscope, and the ratio of the nuclear to plasma of the fibroblasts was large, with unregular nuclear membrane, more Golgi apparatus, rough endoplasmic reticulum, and microfilament and canaliculus appeared. The ultrastructure of the fibroblasts in group B, C, D was spindle and the nuclear was large, with regular nuclear membrane, more Golgi apparatus, rough endoplasmic reticulum. ELISA assay indicated that the secretion of TGF-beta 1 markedly lowered in groups B, C, D in comparison to group A, with the most marked decline observed in group D. Interestingly, we found significantly increased IFN-gamma secretion in groups B, C, D (P < 0.05), with the latter group showing the most notable increase (P < 0.01). Conclusion: These data suggest that both combined and isolated use of CD14 and TLR4 significantly reduce alpha-SMA expression levels, the number of alpha 1 (I) pro-collagen positive cells, and TGF-beta secretion, while substantially increased IFN-gamma secretion. The reduction and increase are especially notable when pretreating with CD14 and TLR4 combined. Here we thus draw a conclusion that both CD14 and TLR4 are associated with the immunophenotype changes and secretion of TGF-beta 1 and IFN-gamma in LPS-stimulated human normal skin fibroblasts.

• 出版日期2015
• 单位中国人民解放军总医院