α晶体蛋白对活化视网膜小胶质细胞iNOS生物活性的影响  

Effects of α-crystailin on bioactivity of iNOS in the activated retinal microglial cells

作　　者：吴楠[1] 徐江宁[1] 李依孺[1] 叶茂[1] 于嘉[1] 王一[1] 

机构地区：[1]第三军医大学附属西南医院眼科,重庆400038

出　　处：《中华创伤杂志》2012年第12期1130-1134,共5页Chinese Journal of Trauma

基　　金：国家自然科学青年基金资助项目（30901645）;第三军医大学青年创新人才基金资助项目（2009XQN29）

摘　　要：目的观察仪晶体蛋白对脂多糖（LPS）诱导活化的视网膜小胶质细胞增生及iNOS生物学活性的影响。方法以离体培养的小胶质细胞为对象，经细胞免疫荧光及流式细胞仪鉴定纯度，用不同浓度LPS和α晶体蛋白干预，MTT法检测α晶体蛋白对视网膜活化小胶质细胞活力的影响；RT—PCR法测定NO浓度，观察小胶质细胞分泌iNOS的表达变化。结果原代培养的小胶质细胞经GSA—IB4免疫组化鉴定及CD11b流式细胞仪鉴定纯度分别达到94．15％和93．34％，10^-4g／Ld晶体蛋白可以抑制10^-6g／LLPS对视网膜小胶质细胞的活力（P〈0．01）；联合用药组iNOS蛋白质量浓度及mRNA表达量明显降低（P〈0．05）。结论在病理情况下，仪晶体蛋白可以通过抑制小胶质细胞产生NO、iNOS，减轻其对视神经损伤后视网膜神经节细胞（retinal ganglial cells，RGCs）的损害。Objective To investigate effects of α-crystallin on proliferation of lipopolysaceharide （LPS）-activated retinal microglia and bioactivity of iNOS. Methods The retinal microglial cells cultured in vitro were analyzed and their purity was identified by cell immunofluorescence and flow cytometry. After mieroglia cells being intervened using LPS and α-crystallin at various concentrations, influence of α-crystallin on activity of LPS-activated retinal microgiia was detected by MTF method and level of NO was measured by RT-PCR to observe changes of iNOS expression in microglia. Results Purity of primary cultured microglial cells was 94.15% by GSA-IB4 immunohistochemical identification and 93.34% by CD11 b flow eytometry. α-crystallin of 10^-4 g/L awakened activity of microglia induced by 10^-6g/L LPS （P 〈0.01 ）. Expressions of iNOS protein and mRNA showed significant decrease in combined treatment group （ P 〈 0.05 ）. Conclusion In clinical condition, α-crystallin decreases the harm of microglial cells on retinal ganglial cells （RGCs） after optical nerve injury by inhibiting the microglia ceils to produce NO and iNOS.

关 键 词：α晶体蛋白质类 小神经胶质细胞 视网膜 细胞培养技术 

分 类 号：R774.6[医药卫生—眼科]