摘要
目的探讨Nogo66蛋白复合物对慢性高眼压大鼠视网膜微环境的影响。方法用532-二极管激光制备慢性高眼压大鼠模型,每只实验大鼠右眼为模型眼,左眼为对照眼。将造模成功的18只大鼠分为两组,分别接种Nogo66蛋白复合物和卵清蛋白复合物(n=9),荧光金逆行标记视网膜神经节细胞(retinal ganglion cells, RGCs)计数,免疫荧光法及免疫组化法观察视网膜GAP43、CD3、脑源性神经营养因子(brain-derived neurotrophic factor, BDNF)、胶质细胞源性神经营养因子(glial cell line-derived neurotrophic factor, GDNF)表达,Western blot检测视网膜BDNF、GDNF蛋白表达。结果慢性高眼压大鼠RGCs计数显著低于对照组大鼠(P<0.01),接种Nogo66-FA复合物后,RGCs数量有所增加,显著高于对照组(P<0.05);且视网膜节细胞层和外丛状层有大量GAP43和CD3表达,视网膜节细胞层、内丛状层和内核层大量表达BDNF和GNDF,视网膜BNDF、GDNF蛋白表达量显著高于对照组(P<0.05)。结论接种Nogo66蛋白复合物后,慢性高眼压模型大鼠视网膜微环境改善,T细胞及多种神经营养因子表达上调,RGCs数量增加。
Objective To investigate the effect of protein complex, Nogo66 and Freund’s adjuvant(FA), on retinal microenvironment in rats with chronic ocular hypertension. Methods After the rat model of chronic high intraocular pressure was established on the right eye(the left eye served as control) with the aid of 532-diode laser, the 18 model rats were equally divided into Nogo66 and ovalbumin groups, receiving vaccination with corresponding agent of 100 μg(in 300 μL FA) on the 1 st, 7 th and 14 th days. The retinal ganglion cells(RGCs) were counted after retrograde labelling with fluorescent gold. The retinal expression of CD3, growth associated protein 43(GAP43), brain-derived neurotrophic factor(BDNF) and glial cell line-derived neurotrophic factor(GDNF) were observed by immunofluorescence assay and immunohistochemical staining, and the protein levels of retinal BDNF and GDNF were detected by Western blotting. Results The count of RGCs was significantly lower in the right eyes with chronic ocular hypertension than the left eyes(P<0.01). Inoculation of Nogo66-FA complex resulted in significantly larger number of RGCs when compared with the rats from the ovalbumin group(P<0.05). In addition, GAP43 and CD3 were highly expressed in the ganglion cell layer and the outer plexus layer, and BDNF and GNDF in the ganglion cell layer, the inner plexus layer and the inner core layer. The protein levels of BNDF and GDNF in the retina were significantly higher in the Nogo66-FA group than the ovalbumin group(P<0.05). Conclusion Inoculation of protein complex Nogo66-FA can improve the retinal microenvironment and the activation of T cells, up-regulate the expression of various neurotrophic factors, and increase the count of RGCs in the rat model of chronic ocular hypertension.
作者
程茗
贺翔鸽
CHENG Ming;HE Xiangge(Department of Ophthalmology,Daping Hospital,Army Medical University(Third Military Medical University),Chongqing,400042,China)
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2021年第7期614-621,共8页
Journal of Third Military Medical University
