金黄色葡萄球菌肽聚糖对HaCaT细胞Toll样受体2和4的影响

目的观察金黄色葡萄球菌胞壁成分肽聚糖对HaCaT细胞Toll样受体(TLR)2和TLR4表达的影响。方法用不同浓度的肽聚糖(10μg/mL、30μg/mL、100μg/mL)处理培养的HaCaT细胞,用RT-PCR检测TLR2 mRNA和TLR4 mRNA含量变化,用流式细胞术检测HaCaT细胞表面TLR2和TLR4蛋白的表达。结果30μg/mL肽聚糖处理组HaCaT细胞的TLR2 mRNA和TLR4 mRNA相对表达水平分别为0.826±0.06和0.801±0.04,明显高于未处理的对照组的TLR2 mRNA(0.433±0.04,P<0.05)和TLR4 mRNA(0.351±0.05,P<0.01)。各浓度的肽聚糖处理HaCaT细胞24h,及100μg/mL浓度的肽聚糖处理HaCaT细胞12h、24h、36h和48h时,及100μg/mL肽聚糖处理24h后,HaCaT细胞表面TLR2和TLR4蛋白表达量均最高。结论肽聚糖可以上调HaCaT细胞TLR2和TLR4的mRNA和蛋白的表达。

The Effect of Peptidoglycan from Staphylococcus Aureus on TLR2 and TLR4 in HaCaT Cells

Objective To study toll-like receptors(TLRs) modulation in HaCaT cells by peptidoglycan from Staphylococcus aureus. Methods HaCaT cells were treated with different concentrations of peptidoglycan(10μg/mL,30μg/mL,100μg/mL),and RT-PCR was used to detect TLR2 mRNA and TLR4 mRNA expression.Flow cytometry was used to assess TLR2 and TLR4 protein expression on surface of HaCaT cells.Results The relative expressions of TLR2 mRNA and TLR4 mRNA of HaCaT cells stimulated with 30μg/mL peptidoglycan in 6 hours were 0.826±0.06 and 0.801±0.04 respectly,which were obviously higher than TLR2 mRNA expression(0.433±0.04,P<0.05 ) and TLR4 mRNA expression(0.351±0.05,P<0.01) in control group.Among cultured with 10μg/mL,30μg/mL,100μg/mL peptidoglycan in 24 hours respectly,and 100μg/mL peptidoglycan in 12, 24,36,48 hours,HaCaT cells stimulated with 100μg/mL peptidoglycan stimulation in 24 hours showed the highest expressions of TLR2 and TLR 4 protein.Conclusion Peptidoglycan can up-regulate both mRNA and protein expressions of TLR2 and TLR 4 in HaCaT cells.