血小板源性生长因子-C蛋白在视网膜光感受器细胞损伤模型中的神经保护作用  

Neuroprotective Effect of PDGF-C Protein in Retina Photoreceptor Damage Models

作　　者：张汝婷 陈伟 胡文婕 唐仲书[1] ZHANG Ru-ting;CHEN Wei;HU Wen-jie;TANG Zhong-shu(Zhongshan Ophthalmic Center,Sun Yat-sen University,Guangzhou 510000,China)

机构地区：[1]中山大学中山眼科中心,广东广州510000

出　　处：《中山大学学报（医学版）》2018年第5期648-653,共6页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：国家自然科学基金(81371021)

摘　　要：【目的】探讨血小板源性生长因子-C(PDGF-C)蛋白治疗对视网膜光感受器细胞损伤模型中的神经保护作用。【方法】(1)检测体外条件下PDGF-C的作用,使用光感受器细胞系661W细胞及原代培养的Pde6brd1视网膜细胞。在体外培养的661W细胞培养液中加入PDGF-C蛋白或者牛血清蛋白(BSA),分别予以同等强度相同时间长短的紫外线照射,然后做MTT检测其细胞凋亡状况,检测PDGF-C蛋白对661W细胞的保护作用。取Pde6brd1小鼠视网膜做原代细胞培养后,在培养液中分别加入PDGF-C蛋白或BSA,相同条件培养后通过免疫荧光染色检测光感受器细胞的生存情况,以检测PDGF-C蛋白对该原代细胞的保护作用。(2)检测在体条件下PDGF-C的作用,在Pde6brd1小鼠的视网膜下分别注射PDGF-C蛋白及BSA,然后通过组织切片后苏木精-伊红(HE)染色观察视网膜光感受器细胞数量的变化来检测PDGF-C的神经保护作用。【结果】在紫外线诱导的视网膜光感受器细胞661W损伤模型中,培养液中加入PDGF-C蛋白的实验组中,MTT实验显示490 nm吸光度(0.2857±0.054)较对照组(0.1726±0.005)增高,P <0.05。在Pde6brd1原代培养的视网膜细胞中,PDGF-C蛋白组中光感受器细胞的存活率明显高于对照组,P <0.05。在Pde6brd1小鼠的视网膜下注射PDGF-C蛋白,实验组外核层中的每100μm细胞计数(38.4769±6.1)较对照组(27.2435±5.3)增加,P <0.05。【结论】PDGF-C蛋白在光感受器细胞损伤模型中具有较强的神经保护作用。【Objective】We investigated the neuroprotective effect of Platelet-derived growth factor C(PDGF-C)pro-tein in retina photoreceptor cell damage models.【Methods】Both in vivo and in vitro models have been tested.One is UVlesioned photoreceptor cell line 661W.Neuroprotective effect of PDGF-C was assessed by MTT test.Another model is a typical retinitis pigmentosa model called Pde6brd1.Neuroprotective effect of PDGF-C protein was assessed by immunos-taining through retina cell primary culture and HE staining through subretinal injection.【Result】Neuroprotective effect is detected in UV-lesioned 661W cell model.The results of MTT test showed that the absorbance values in PDGF-C groups increased obviously compared with control.Furthermore,immunostaining results showed that survival of photoreceptor cells increased in PDGF-C treated group compared with control.Moreover,subretinal injection of PDGF-C increased the cell number in outer nuclear layer.【Conclusion】Thus,the cell and animal models valicated the neuroprotective effect of PDGF-C in photoreceptor lesions.

关 键 词：血小板源性生长因子-C rd1 光感受器细胞 661W细胞 

分 类 号：R453.9[医药卫生—治疗学]