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Co-regulation of Dectin-1 and TLR2 in inflammatory response of human corneal epithelial cells induced by Aspergillus fumigates
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AIM: To investigate the co-regulation of dendritic cell-associated C-type lectin-1 (Dectin-1), Toll-like receptor 2 (TLR2), and relative chemotactic factors in the Telomease-immortalized human corneal epithelial (THCE) cells after exposure to Aspergillus fumigatus (Af) hyphae.
METHODS: The normal THCE cells were investigated as control. After cultured in vitro with Af hyphae, with or without laminarin and anti-TLR2 antibody for 4, 8, 16 and 24h, THCE cells were harvested. The expression of Dectin-1, TLR2, CXCL1 and CXCL8 mRNA were measured by real-time quantitative polymerase chain reaction at the stimulation of 4, 8 and 16h separately. The protein expression of Dectin-1 and TLR2 were analyzed at 8, 16, and 24h by Western blot.
RESULTS: The mRNA expression of CXCL1 and CXCL8 increased in THCE cells after stimulated by Af hyphae. The stimulatory effects on these inflammatory chemokines were shown in a dose-dependent manner and reached the peak at 8h. Af hyphae significantly stimulated the production of Dectin-1 and TLR2 in THCE cells at both mRNA and protein levels. The protein of Dectin-1 and TLR2 gradually increased till 16h. While pretreated with laminarin (a Dectin-1 inhibitor), the expression of TLR2, CXCL1 and CXCL8 all decreased dramatically at the peak point. Interestingly, when pretreated with TLR2 neutralizing antibody, the expression of Dectin-1, CXCL1 and CXCL8 also decreased dramatically at the peak point.
CONCLUSION: These findings suggest that Dectin-1 and TLR2 co-regulated with each other after treated with inactive Af hyphae in the THCE cells, and they contribute together to the inflammatory responses by induction of chemokines CXCL1 and CXCL8. 相似文献
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目的:探讨基质金属蛋白酶-2(matrix metalloproteinase-2,MMP-2)及其抑制剂(tissue inhibitor of metalloproteinase-2,TIMP-2)在大鼠外伤性白内障形成过程中所发挥的作用。方法:正常成年SD大鼠20只,体质量250~300g,建立单眼晶状体穿通伤后外伤性白内障模型,并随机分为伤后12,24,72,168h4个时相组。用免疫组织化学染色法检测MMP-2和TIMP-2在各组晶状体上皮细胞(lens epithelial cell,LEC)中的表达情况。结果:外伤后12h晶状体开始出现混浊,并随时间延长而加重。MMP-2在各外伤组LEC中均有较强的阳性表达,而在正常对照眼中无表达。TIMP-2的表达在各组LEC中差异均无显著意义(P>0.05)。MMP-2与TIMP-2阳性表达率间无相关性(P>0.05)。结论:MMP-2参与了外伤性白内障形成的病理环节。MMP-2和TIMP-2表达失衡在外伤性白内障的发生和发展过程中发挥重要作用。 相似文献
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枸杞多糖对氧化损伤大鼠晶状体上皮细胞凋亡相关基因Bcl-2和Bax表达的调控 总被引:1,自引:0,他引:1
目的:研究枸杞多糖(lycium barbarum polysaccha-rides,LBP)对实验性氧化损伤大鼠晶状体上皮细胞(lens ep- ithelial cell,LEC)凋亡相关基因Bcl-2和Bax表达的调控。方法:复制大鼠晶状体氧化损伤模型,同时加入终浓度为1g/L的枸杞多糖共同孵育24h后,取晶状体前囊膜采用免疫组化法检测凋亡相关基因Bcl-2和Bax的蛋白表达。结果:Bcl-2和Bax在正常SD大鼠LEC中均有表达,Bcl-2的表达远较Bax表达强。H_2O_2组Bcl-2表达显著下调,Bax表达显著上调(Ridit检验,R_1值分别为0.97和0,P<0.01,差异均有非常显著性),Bcl-2/Bax比率下降。枸杞多糖可明显上调Bcl-2表达,下调Bax的表达(与H_2O_2组比较,P值均<0.01,差异有非常显著性),Bcl-2/Bax比率上升;其改变幅度与白内停组相比差异均有显著性(P值均<0.05)。结论:氧化损伤诱导LEC凋亡和枸杞多糖抑制LEC凋亡的分子机制可能与调控凋亡相关基因Bcl-2和Bax的表达有关。 相似文献