摘要
目的探讨局部应用CpG ODN对小鼠实验性变态反应性结膜炎的治疗作用和相关机制,以期为变态反应性结膜炎提供新的治疗策略。方法随机将24只6~8周雌性Balb/c小鼠分为4组：CpG ODN组、GpC ODN组、PBS组和正常对照组,每组各6只,鸡卵蛋白（OVA）免疫Balb/c小鼠建立变态反应性结膜炎动物模型,首次激发诱导变态反应性结膜炎成功后第4天双眼连续3d分别滴入CpG ODN（5g·L-1）、GpC ODN（5g·L-1）和PBS各10μL,治疗后的第4天观察眼部临床症状后处死动物,检测结膜组织切片中嗜酸性粒细胞和中性粒细胞的变化、血清中IgG1、IgG2a含量和细胞分泌IL-4的水平,流式细胞仪对培养细胞的分子表达和含量进行分析。结果与GpC ODN对照组和PBS对照组比较,CpG ODN治疗组的主要指标变化为：（1）结膜组织切片中嗜酸性粒细胞和中性粒细胞的数量明显减少,嗜酸性粒细胞每张切片上的细胞数分别为：CpG ODN组（10.00±0·92）个,GpC ODN组（17.00±1.06）个,PBS组（20.00±1·26）个。中性粒细胞每张切片上的细胞数分别为：CpG ODN组（15.00±1.36）个,GpC ODN组（32.00±2.83）个,PBS组（29.00±1.58）个;（2）血清中IgG2a的含量增加,IgG1/IgG2a比例下降（P〈0·01）;（3）细胞培养上清中的IL-4分泌下调,CpG ODN组（20.702±1.688）ng·L-1,GpC ODN组（27.201±1·274）ng·L-1,PBS组（30.534±2.576）ng·L-1;（4）CD4＋CD25＋和CD11c＋CD80＋MHCⅡ＋的细胞比例有所增加,其中CD4＋CD25＋细胞所占比例在CpG ODN组为9·55%,GpC ODN组为5.00%,PBS组为5.92%.CD80和MHCⅡ＋共表达的细胞占CD11c＋细胞比例在CpG ODN组为5.71%,GpCODN组为3.51%,PBS组为2.18%.但CpGODN治疗组对变态反应性结膜炎早期临床症状的改善没有明显作用（P〉0·05）。结论眼部应用CpG ODN能够逆转实验性变态反应性结膜炎小鼠激发阶段体内的Th2型应答,改变血清中抗体亚型,减轻结膜组织局部的晚期病理反应,对实
Objective To investigate the therapeutic effect of topical CpG ODN treatment on experimental allergic conjunctivitis and provide new treatment policies.Methods Twenty-four female Balb/c mice at the age of six to eight weeks were divided into four groups randomly：CpG ODN group,GpC ODN group,PBS group and normal control group,six in each group.Allergic conjunctivitis models were set up with OVA-induced Balb/c mice.CpG ODN （5 g·L-1,10 μL） were applied topically for three consecutive days at the fourth day after successfully induced AC.GpC ODN and PBS were 10 μL,respectively.Four days after the last treatment,the clinical symptoms were examined before executing the mice.Changes of eosinophil and neutrophil in conjunctival tissue slice,content of IgG2a and IgG1 in serum and the level of IL-4 were measured.Molecular expression and content of cultured cells were analyzed.Results Compared with GpC ODN and PBS groups,CpG ODN group manifested lower inflammation cell counts,and average eosinophil number in each section were （10.00±0.92）,（17.00±1.06） and （20.00±1.26） respectively;Average neutrophil count in each section were （15.00±1.36）,（32.00±2.83） and （29.00±158） respectively.There were higher IgG2a production in serum（P〈0.01）,and lower IL-4 level：（20.702±1.688）ng·L-1 in CpG ODN group,（27.201±1.274）ng·L-1 in GpC ODN group and （30.534±2.576）ng·L-1 in PBS group.In cultured splenocytes supernatant,there were increased cell percentage of CD4＋CD25＋（9.55% in CpG ODN,5.00% in GpC ODN and 5.92% in PBS） and CD11c＋CD80＋MHCⅡ＋ （5.71% in CpG ODN,3.51% in GpC ODN and 2.18% in PBS） in cultured splenic cells.However,the differences of clinical scores among each group were not statistically significant（P〉0.05）.Conclusion Topical CpG ODN treatment might inhibit late-phase reaction of allergic conjunctivitis,possibly by induction of IgG subclass switching and modulation of Treg and dendritic cells response in treated allergic conjunctivitis mice.It
出处
《眼科新进展》
CAS
2008年第3期172-176,共5页
Recent Advances in Ophthalmology
基金
国家自然科学基金资助(编号:30772375)~~
