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目的研究羊膜对兔视网膜Mller细胞表皮生长因子受体(EGFR)表达的影响及其机制。方法取出生15d新西兰白兔视网膜Mller细胞进行原代培养及传代,采用神经胶质纤维酸性蛋白(GFAP)和S-100进行细胞鉴定。0.25%胰蛋白酶、0.05%EDTA酶消化取第3代Mller细胞,实验组加0.5mL羊膜匀浆上清液继续培养12h,采用免疫组织化学法检测羊膜诱导和非诱导条件下兔视网膜Mller细胞EGFR表达的变化并进行分析。结果培养的兔视网膜Mller细胞GFAP和S-100表达阳性,透射电镜检查细胞质内可见8～10nm的中间丝。未经羊膜诱导的视网膜Mller细胞EGFR有少量表达,经羊膜诱导12h后,EGFR表达明显增多,2组灰度值分别为571588.80±67862.68和1000352.00±98386.22,差异有统计学意义(t=4.035,P<0.01)。结论经羊膜诱导后可促进培养的兔视网膜Mller细胞EGFR的表达。
Objective To study the effect of amniotic membrane on the expression of epidermal growth factor receptor (EGFR) in retinal Mäller cells in rabbits and its mechanism. Methods Rabbit retina Mller cells from New Zealand white rabbits were primarily cultured and passaged for 15 days. GFAP and S-100 were used for cell identification. 0.25% trypsin, 0.05% EDTA digestion took the third generation Mller cells, the experimental group plus 0.5mL amniotic homogenate supernatant was cultured for 12h, Immunohistochemistry was used to detect amniotic membrane in rabbit retinal M The changes of EGFR expression in ller cells were analyzed and analyzed. Results The cultured rabbit retinal Mller cells showed positive GFAP and S-100 expression, and the intermediate filaments with 8 ~ 10 nm were observed in the cytoplasm by transmission electron microscopy. There was a small amount of EGFR expression in retinal Mller cells without amnion, and the expression of EGFR increased significantly after 12h of amnion induction. The gray values of two groups were 571588.80 ± 67862.68 and 1000352.00 ± 98386.22 respectively, with significant difference (t = 4.035 , P <0.01). Conclusion Amniotic membrane can promote EGFR expression in cultured rabbit retinal Mller cells.