羊膜对体外培养的兔视网膜Müller细胞表皮生长因子受体表达的影响

目的研究羊膜对兔视网膜Müller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Müller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Müller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Müller细胞EGFR表达的变化并进行分析。结果培养的兔视网膜Müller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Müller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Müller细胞EGFR的表达。

The effects of amnion on expression of EGFR in rabbit M(u)ller cells

Objective hole.Research demonstrated that amnion promotes the proliferation of and its mechanism.Methodswhite rabbit.The cells were primarily cultured and passaged in DM/F12 with 10% fetal bovine serum and identified with glial fibrillary acidic using 0.25% trypsin and 0.05%EDTA and suspended in 10% fetal bovine serum+DMEM/F12.0.5 mL of the amnion supernatant was added to the medium for 12 hours.The expression of EGFR in the cells was detected by immunohistochemistry.The difference of EGFR expression with or without the amnion was evaluated.ResultsCultured cells showed a positive response for GFAP and S-100.The intermediate filaments with a length of 8-10 nm were exhibited under the transmission electron microscope.EGFR was fainly expressed in cultured hours,the expression of EGFR was obviously increased with a significant difference in the gray scale between the two groups(571 588.80±67 862.68 vs.1 000 352.00±98 386.22)(t=4.035,P＜0.01).Conclusion