与胶原纤维的作用是Mller细胞黏附于玻璃体的主要机制

Interaction with collagen fiber is the main mechanism of Mller cell adhesion to vitreous gel

目的:观察Mller细胞在玻璃体凝胶上的黏附并分析其机制。方法:猪视网膜Mller细胞提取培养并以酸性纤维蛋白、波形蛋白和谷氨酰胺合成酶免疫荧光染色确认,分别与抗细胞表面胶原受体整联蛋白α2β1抗体、抗透明质酸受体CD44抗体培养后,播于新鲜玻璃体上培养2 h,以PBS冲洗3次后计数剩余细胞数目,并与未用抗体培养的对照组相比较。结果:所提取细胞97.7%胶质纤维酸性蛋白染色阳性,94.6%波形蛋白染色阳性,86.0%谷氨酰胺合成酶染色阳性。细胞在玻璃体上培养2 h后开始附着于玻璃体,培养72 h以后细胞增殖形成网状结构并进入玻璃体凝胶中;抗整联蛋白α2β1抗体可使剩余细胞较对照组显著减少,抗CD44抗体不产生明显差异。结论:Mller细胞可黏附于玻璃体上,增殖并进入玻璃体凝胶,其表面胶原受体与玻璃体Ⅱ型胶原纤维的作用是其附着于玻璃体的主要机制。

Objective：To investigate the mechanism of Mller cell adhesion to vitreous gel.Methods：Porcine Mller cells were extracted and cultured,identified with immuno-recognition with glial fibrillary acidic protein（GFAP）,vimentin and glutamine syn-thetase（GS） antibody.Cells were collected and incubated with / without integrinα2β1 and CD44 antibody before application to fresh porcine vitreous for 2 hours,and then washed with PBS for 3 times.Remaining cells on the vitreous were counted.Results： ① Cells collected were 97.7% positive for GFAP,94.6% positive for vimentin,86.0% positive for GS,respectively.Cells adhered to vitreous after 2 hours of culture,formed net-like structure and enter vitreous gel after 72 hours;② Pre-incubation with integrinα2β1 antibody significantly decreased the remaining cell,while no such significance was observed in the cells incubated with CD44 antibody.Con-clusion：Mller cell can adhere to vitreous and proliferate,in which the interaction with type Ⅱ collagen is an important mechanism.