压力对大鼠纯化视网膜神经节细胞中神经微丝的影响

Effects of Pressure on the NF-H of Rat Retinal Ganglion Cells

目的研究压力对体外培养的视网膜神经节细胞(retinal ganglion cells,RGCs)中神经微丝的影响,探讨高眼压性青光眼中细胞骨架蛋白的表达变化。方法建立体外加压培养纯化的Sprague-Dawley(SD)乳鼠视网膜神经节细胞模型,采用抗大鼠Thy1.1单克隆抗体免疫荧光细胞化学染色法鉴定RGCs的纯度。将纯化培养的SD乳鼠RGCs分别在0、20、40、60及80 mmHg的压力下培养,免疫组织化学方法检测神经微丝蛋白-H(NF-H)的表达,并用图像分析系统对各组NF-H表达进行分析测定。结果①纯化后的视网膜神经节细胞纯度为96%。②压力为20 mmHg时,实验组RGCs中NF-H表达与对照组相比,差异无统计学意义(P>0.05),且NF-H在视网膜神经节细胞中的分布相对均一;压力为40、60及80 mmHg时,实验组RGCs中NF-H表达与对照组相比差异均有统计学意义(均P<0.01),且NF-H在视网膜神经节细胞中分布紊乱。结论纯化的RGCs可以在体外存活并生长,一定的压力(≥40 mmHg)可影响RGCs中细胞骨架蛋白的表达。

Objective To study neurofilament protein expression in rat purified retinal ganglion cells（RGCs） that cultured under different pressures. Methods The hyperbaric model of cultured and purified SD suckling rats＇ RGCs in vitro was established. The purity of RGCs was identified by immunofluorescent cytochemistry with anti-rat Thy1. 1 monoclonal antibody. The RGCs were cultured under pressures of 0,20,40,60 and 80 mmHg,respectively. The expression of NF-H protein in RGCs under different pressures was detected quantitatively by immunohistochemistry. Imaging analysis system was used to analyze the expression of NF-H. Results① After cultured for 12 h in vitro,the purity of RGCs in the experiment was 96 %. ② If the pressure was equal to or higher than 40 mmHg,the expression of NF-H in RGCs became weaker and weaker as the pressure was increased,and was very significantly different from that in control group（P〈0.01）. If the pressure was lower than 20 mmHg, the expression of NF-H in RGCs was similar to that of the control group（P〉0.05）. Conclusion Purified RGCs can survive and grow in vitro. Certain pressures（≥40 mmHg） have strong effects on the cytoskeletal protein of RGCs cultured in vitro.