α-crystallin与多种细胞作用的免疫荧光观察

Immunofluorescence study of interaction of α crystallin and many cells

目的在α-crystallin(α晶体蛋白)对RGCs保护性作用研究并进行定位研究的基础上,对α-crystallin与其他细胞之间的作用相比较,以期揭示α-crystallin与RGCs作用的本质。方法原代培养大鼠视网膜神经节细胞(RGCs)、大鼠巨噬细胞、大鼠心肌细胞以及传代培养的ECV细胞株、293细胞细胞株、猪视网膜色素上皮细胞(RPE)。应用免疫荧光结合激光共聚焦技术观察α-crystallin与多种细胞的作用。结果免疫荧光结合激光共聚焦发现α晶体蛋白在3种细胞中有阳性免疫沉积物出现在细胞膜上,其对照组阴性;3种细胞见实验组和对照组均有阳性免疫沉积物出现在细胞浆、细胞核中。结论α晶体蛋白不是特异性与RGCs发生作用,它是广泛存在的生物学效应物质。α晶体蛋白具有s HSP的抗应激损伤特性,对于不表达内源性α晶体蛋白的同时又是抗损伤能力较弱的RGCs,可通过增加外源性α晶体蛋白达到有效保护RGCs的目的。

Objective To study the interaction phenomen of α-crystallin and retinal ganglion cells（RGCs） ,other cells such as 293 cells,ECV cells, retinal pigment epithilial cells （RPE）, myocardial cell and macrophage were observed too. Methods RGCs, 293 cells,ECV cells, retinal pigment epithilial cells, myocardial cell and macrophage were cultured respectively. The interaction phenomen of α-crystallin and other cells was detected by immunofluorescence with confocal laser scanning microscope. Results The interaction between a crystallin and RGCs was detected positive by immunofluorescence with confocal laser scanning microscope. The results about interaction phenomen of α-crystallin and 293 cells, ECV cells were as same as that and RGCs, while interaction of α- crystallin and retinal pigment epithilial cell,myocardial cell and macrophage were different from that of α-crystallin and RGCs,They were detected positive with immunofluorescence staining with α-crystallin antibody even not interacting with α-crystallin. Conclusion a crystallin can interact with RGCs,the act point is on the membrane of RGCs. A-crystallin has not only action on RGCs but also on other cells. We can add ectogenesis α-crystallin to protect RGCs when they were injuried.