金黄色葡萄球菌对人角质形成细胞Toll样受体2及β-防御素-3影响研究

Human keratinocytes infected with staphylococcus aureus,the changes of toll-Iike receptor 2 and beta-defensin-3 in cells

目的观察金黄色葡萄球菌侵染人角质形成细胞(HaCaT细胞)对胞内Toll样受体2(TLR2)和β-防御素-3(hBD-3)表达的影响。方法设空白对照组和MOI(感染复数=细菌量∶细胞量)分别为1∶20,1∶10,1∶1,3∶1,10∶1,20∶1,50∶1,100∶1和200∶1的金葡菌处理组,用CCK-8检测细胞活力,选择细胞死亡过半组的MOI=20∶1作为最适菌种浓度。复设实验对照组和MOI=20∶1染菌实验组,RT-PCR检测胞内TLR2和hBD-3 mRNA的表达水平。结果与空白对照组比较,MOI为20∶1菌种浓度处理HaCaT细胞8h细胞死亡过半,随着菌种浓度的增大,细胞活力逐渐减弱。染菌HaCaT细胞中TLR2 mRNA、hBD-3 mRNA的表达均增高(P<0.05)。结论金葡菌感染可造成人角质细胞活力减弱,胞内TLR2及hBD-3的表达增高。

Objective To investigate the effects of staphylococcus aureus（ s. aureus） on the expression of intracellular toll-like receptor 2 and beta-defensin-3 in human keratinocytes. Methods The blank control group and s.aureus treatment group were set up. The MOI（ multiplicity of infection = bacterial count∶ cell volume） of s. aureus treatment group was divided into 1∶ 20,1∶ 10,1∶ 1,3∶ 1,10∶ 1,20 ∶ 1,50 ∶ 1,100 ∶ 1,and 200 ∶ 1. CCK8 was used to detect the cell vitality. The optimal concentration of MOI = 20∶ 1 in the cell death group was chosen as the optimal concentration. The blank control group and MOI = 20 ∶ 1 group were reconstituted. The expression levels of intracellular TLR2 and hBD-3 mRNA were detected by RT-PCR. Results Compared with the blank control group,the concentration of HaCaT cells in the MOI = 20∶ 1 group was more than half of the death. As the concentration of bacteria increased,the cell vitality gradually decreased. The expression of TLR2 mRNA and hBD-3 mRNA in HaCaT cells was both increased（ P〈0. 05）. Conclusion The infection of s. aureus can cause a decrease in the vitality of keratinocytes,and the expression of intracellular TLR2 and hBD-3 is increased.