不同浓度肽聚糖对角膜上皮细胞Toll样受体(TLR)2和TLR4表达的影响

目的 研究不同浓度肽聚糖对角膜上皮细胞Toll样受体(Toll-like receptor,TLR)2和TLR4表达的影响.方法 原代培养C57小鼠角膜上皮细胞,随机分为对照组(无肽聚糖刺激)及按照肽聚糖刺激浓度分为10 mg·L-1组、30 mg·L-1组、80 mg·L-1组(作用时间均为12 h);按照肽聚糖刺激时间分为12 h组、24 h组、36 h组(作用浓度均为30mg·L-1),应用实时荧光定量PCR与流式细胞术检测各组TLR2、TLR4 mRNA和蛋白的表达量.结果 与对照组(1.00±0.14、1.00±0..01)相比,10 mg·L-1组(4.35±0.46、3.53±0.50)、30 mg· L-1组(8.06±0.72、5.31±0.34)、80 mg· L-1组(2.93±0.46、2.23±0.04)的TLR2、TLR4 mRNA表达增加,差异均有统计学意义(均为P＜0.05);与对照组(1.00±0.14、1.00±0.01)相比,12h组(2.94±0.46、2.23±0.50) TLR2、TLR4 mRNA表达增加,差异均有统计学意义(均为P＜0.05).与对照组相比,各浓度组和12 h组角膜上皮细胞TLR2、TLR4蛋白表达均增加,差异均有统计学意义(均为P＜0.05).结论 肽聚糖可以刺激小鼠角膜上皮细胞TLR2、TLR4 mRNA和蛋白表达增加,提示感染性角膜炎的发生、发展可能与角膜上皮细胞TLR2、TLR4识别病原体并参与炎症反应有关.

Effects of peptidoglycan with different concentrations on TLR2 and TLR4 in corneal epithelial cells of mice

Objective To investigate the effects of peptidoglycan (PGN) with different concentrations on Toll-like receptor 2 (TLR2),Toll-like receptor 4 (TLR4) expression in corneal epithelial cells of mice.Methods Corneal epithelial cells of c57 mice were cultured in vitro.Cells were divided into blank control group and 10 mg · L-1 group,30 mg · L-1 gruop and 80 mg · L-1 group (treated by different concentration of PGN for 12 hours).In the meantime,the cells in 30 mg · L-1 group were cultured for different times(named 12 hours group,24 hours group,36 hours group).Expressions of TLR2 and TLR4 mRNA and protein in different group were measured by RT-PCR and flow cytometry.Results Compared with control group (1.00 ± 0.14,1.00 ± 0.01),the expression of TLR2,LR4 mRNA in 10 mg · L-1 group (4.35 ± 0.46,3.53 ± 0.50),30 mg · L-1 group (8.06 ±0.72,5.31 ±0.34),80 mg · L-1 group (2.93 ±0.46,2.23 ±0.04) were increased,the differences were statistically significant (all P ＜ 0.05).Compared with control group,the expression of TLR2,TLR4 protein in different concentration group and 12 hour group were increased,the differences were statistically significant (all P ＜ 0.05).Conclusion PGN can up-regulate both mRNA and protein expression of TLR2 and TLR4 in corneal epithelial cells of mice,suggest that TLR2 and TLR4 in the corneal epithelial cell can recognize some exogenous pathogen and regulate the inflammatory reaction,which are closely related to the occurrence and development of infectious keratitis.