[1]刘晶,白浪,苏亚茹,等.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[J].眼科新进展,2017,37(8):705-708.[doi:10.13389/j.cnki.rao.2017.0179]
　LIU Jing,BAI Lang,SU Ya-Ru,et al.Effects of peptidoglycan with different concentrations on TLR2 and TLR4 in corneal epithelial cells of mice[J].Recent Advances in Ophthalmology,2017,37(8):705-708.[doi:10.13389/j.cnki.rao.2017.0179]
点击复制

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/HTML
分享到：

《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
37卷
期数:
2017年8期
页码:
705-708
栏目:
实验研究
出版日期:
2017-08-05

文章信息/Info

Title:
Effects of peptidoglycan with different concentrations on TLR2 and TLR4 in corneal epithelial cells of mice
作者:
刘晶; 白浪; 苏亚茹; 于健; 孟婷; 陈敏婷
510000　广东省广州市，南方医科大学南方医院眼科
Author(s):
LIU Jing; BAI Lang; SU Ya-Ru; YU Jian; MENG Ting; CHEN Min-Ting
Department of Ophthalmology,Nanfang Hospital of Southern Medical University,Guangzhou 510000,Guangdong Province,China
关键词:
肽聚糖; Toll样受体家族; 角膜上皮细胞; 感染性角膜炎
Keywords:
peptidoglycan; Toll like receptor 2; Toll like receptor 4; infectious keratitis
分类号:
R772.2
DOI:
10.13389/j.cnki.rao.2017.0179
文献标志码:
A
摘要:
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30 mg·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，12 h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。
Abstract:
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,TLR4 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.

参考文献/References:

［1］　杨杰,汪怿,温积权,季娴,刘健,江媛.感染性角膜炎患者的病原菌分布与耐药性分析［J］.中华医院感染杂志,2016,26(19):4497-4499.
YANG J,WANG Y,WEN JQ,JI X,LIU J,JIANG Y.Pathogen distribution and drug residtance of infectious keratitis［J］.Chin J Nosocomiol,2016,26(19):4497-4499.
［2］　EVANS DJ,MCNAMARA NA,FLEISZIG SM.Life at the front:dissecting bacterial-host interactions at the ocular surface［J］.Ocul Surf,2007,5(3):213-227.
［3］　HUANG LC,JEAN D,PROSKE RJ,REINS RY,MCDERMOTT AM.Ocular surface expression and in vitro activity of antimicrobial peptides［J］.Curr Eye Res,2007,32(7-8):595-609.
［4］　LAMBIASE A,MICERA A,SACCHETTI M,MANTELLI F,BONINI S.Toll-like receptors in ocular surface diseases:overview and new findings［J］.Clin Sci,2011,120(10):441-450.
［5］　WONG Y,SEHU C,LOUAFI F,HOSSAIN P.Lipopolysaccharide regulation of toll-like receptor-4 and matrix metalloprotease-9 in human primarycorneal fibroblasts［J］.Invest Ophthalmol Vis Sci,2011,52(5):2796-803.
［6］　SVEDOVA J,TSURATANI N,LIU W,KHANNA KM,VELLA AT.TNF and CD28 signaling play unique but complementary roles in the systemic recruitment of innate immune cells after staphylococcus aureus enterotoxin a inhalation［J］.J Immunol,2016,196(11):4510-4521.
［7］　袁钊辉,龙崇德,林晓峰,张铁英,娄秉盛,刘奕志.金黄色葡萄球菌胞壁成分在细菌性眼内炎中的致病作用［J］.中国病理生理杂志,2010,26(10):1901-1907.
YUAN ZH,LONG CD,LIN XF,ZHANG TY,LOU BS,LIU YZ.Bacterial cell wall components of staphylococcus aureus induce thalmitis［J］.Chin J Pathophys,2010,26(10):1901-1907.
［8］　苏亚茹,白浪,刘晶,梁伟怡,于健,孟婷.TLR2调节Th细胞极化对小鼠角膜移植排斥反应发生的影响［J］.眼科新进展,2016,36(7):601-604.
SU YR,BAI L,LIU J,LIANG WY,YU J,MENG T.Effects of Th cell polarization regulated by TLR2 on immune rejection after corneal transplantation inmouse［J］.Rec Adv Ophthalmol,2016,36(7):601-604.
［9］　HAMERMAN JA,POTTLE J,NI M,HE Y,ZHANG ZY,BUCKNER JH.Negative regulation of TLR signaling in myeloid cells-Implications for autoimmune diseases［J］.Immunol Rev,2016,269(1):212-227.
［10］　RREDFERN RL,PATEL N,HANLON S,FARLEY W,GONDO M,PFLUGFELDER SC,et al.Toll-like receptor expression and activation in mice with experimental dry eyeTLRs and AMPs in EDE［J］.Invest Ophthalmol Vis Sci,2013,54(2):1554-1563.
［11］　LANG LL,WANG L,LIU L.Exogenous MD-2 confers lipopolysaccharide responsiveness to human corneal epithelial cells with intracellular expression of TLR4 and CD14［J］.Inflammation,2011,34(5):371-378.
［12］　刘苏俊,林麟,周武庆,张彩萍,冯雨苗,马一平.金黄色葡萄球菌肽聚糖对HaCaT细胞TLR2和4的影响［J］.中国皮肤性病学杂志,2009,23(2):84-86.
LIU SJ,LIN L,ZHOU WQ,ZHANG CP,FENG YM,MA YP.The effect of peptidoglycan from staphylococcus aureus on TLR2 and TLR4 in hacat cells［J］.Chin J Derm Venereol,2009,23(2):84-86.
［13］　CAO L,YANG XJ,LI ZJ,SUN FF,WU XH,YAO JH.Reduced lesions in chickens with Clostridium perfringens-induced necrotic enteritis by Lactobacillus fermentum 1.2029［J］.Poult Sci,2012,91(12):3065-3071.
［14］　BOTOS I,SEGAL DM,DAVIES DR.The structural biology of Toll-like receptors［J］.Structure,2011,19(4):447-459.
［15］　REDGERN RL,MCDERMOTT AM.Toll-like receptors in ocular surface disease［J］.Exp Eye Res,2010,90(6):679-687.
［16］　LI Y,YANG H,WU X.Pretreatment with TLR2 and TLR4 ligand modulates innate immunity in corneal fibroblasts challenged with Aspergillus fumigatus［J］.Invest Ophthalmol Vis Sci,2013,54(6):4261-4270.

备注/Memo

备注/Memo:
国家自然科学基金资助（编号:81170887）；南方医科大学南方医院横向课题基金资助（编号：G201202）
更新日期/Last Update: 2017-08-14