压力对视网膜神经节细胞存活及突起生长影响的研究

Effect of pressure on the survival and process growing of retinal ganglion cells

目的 建立体外加压培养SD乳鼠视网膜神经节细胞 (RGCs)模型 ,并观察压力对RGCs存活及突起生长状况的影响。方法 取生后 0～ 3天SD乳鼠的视网膜组织 ,用酶化学方法制备成细胞悬液并接种于培养瓶中。用自行设计的体外加压模型 ,使瓶内压力分别达到 2 0mmHg、4 0mmHg、6 0mmHg和 80mmHg,同时设对照组 (压力为 0 )。分别于培养后 2 4h、4 8h和 72h在倒置显微镜下观察RGCs的存活及生长状况 ,并应用Thy 1 .1单克隆抗体对RGCs进行免疫细胞化学鉴定。结果 压力为 2 0mmHg及 4 0mmHg实验组中RGCs生长状况与对照组相似 ,RGCs存活数量及突起生长率无显著性差异 (P >0 .0 5 ) ;压力为 6 0mmHg及 80mmHg实验组中RGCs存活数量及突起生长率与对照组存在显著性差异 (P <0 .0 5 )。结论 RGCs可以在体外存活并生长 ,一定的压力 (≥ 6 0mmHg)可以影响RGCs的存活数量及突起生长率。

Objective To establish the model of cultivation of SD suckling rats' retinal ganglion cells(RGCs) in vitro underpressure and observe the effect of pressure on them.Methods Zero to three day postnatal SD suckling rats' retinal tissue was dissected and digested with trypsine to make cell suspension.Then the cell suspension was planted into culture flask and injected air into it.The pressure remained at 20 mmHg, 40 mmHg ,60 mmHg,80 mmHg in the flask and 0 in control group.The cells' surviving and living conditions were observed 24,48,72 hours later and RGCs were identified with Thy 1.1 monoclonal antibody.Results The surviving and living conditions of RGCs in 20 mmHg and 40 mmHg experiment groups were similar with that of the control group ( P >0.05),but differences between 60 mmHg,80 mmHg experiment groups and control group were significant statistically ( P <0.05).Conclusion Certain pressure (≥60mmHg) have strong effect on the surviving and living conditions of RGCs cultivated in vitro .