【摘要】 目的 观察正常SD大鼠暗适应视网膜电图b波光强—振幅曲线的特点。方法 取完全暗适应状态SD大鼠12只。利用中性滤光片降低全视野刺激器固有光强度,按0.5个对数单位的间隔衰减得到7档(分别为-5.0、-4.5、-4.0、-3.5、-3.0、-2.5和-2.0 log cd·s·m-2),采用RETI-scan系统,自制的银环角膜电极和不锈钢针状电极,记录暗适应系列光视网膜电图。采用Naka-Rushton方法分析b波的强度—反应函数。结果 b波阈强度为-4.5 log cd·s·m-2,饱和强度为-2.5 log cd·s·m-2。当刺激强度在-5.0~-2.0 log cd·s·m-2范围时,b波光强—振幅曲线呈“S”型,符合Naka-Rushton等式;其中有两个重要参数(Rmax,I50)分别代表b波饱和幅值和产生1/2该幅值的刺激强度,本实验中Rmax为(721.2±89.5)μV,I50为(-3.30±0.36)log cd·s·m-2。上述参数与人类正常值相比差异明显。结论 大鼠视网膜反应特性与人类存在较大差异,应用视网膜电图b波光强—反应函数来评价其杆体系统功能时选择合适的刺激参数非常重要。
【关键词】 视网膜电图;SD大鼠;光强—反应函数
The luminance response function of the electroretinogram of the dark-adapted SD rat
DAI Xufeng*, LIU Xiaoling*, ZHAO Huiling.
School of Ophthalmology and Optometry, Wenzhou Medical College, Wenzhou China, 325027
[Abstract] Objective To investigate the b-wave luminance response function of the scotopic electroretinogram in the normal SD rat. Methods Twelve dark-adapted SD rat eyes were included in the experiment. An original Ganzfeld light was spectrally filtered with a 500-nm interference filter to produce flashes that varied in intensity from -5.0~-2.0 log cd·s·m-2. ERGs to a series of flashes ranging in the intensities mentioned above were recorded with a RETI port system, silver loop electrode and stainless steel needle electrode. The Naka-Rushton equation was used to describe the b-wave luminance response function. Results The intensity that provided the b-wave threshold response and saturation measured -4.5 and -2.5 log cd·s·m-2, respectively. The b-wave amplitudes were plotted as an S-shaped curve with an increase in intensity from -5.0 to -2.0 log cd·s·m-2. The relationship between them could be plotted using a hyperbolic saturation function (Naka-Rushton function). The model yielded 2 often-used parameters, Rmax and scotopic I50, representing the asymptotic b wave amplitude and the intensity that provided half saturation. In this experiment, Rmax measured (721.2±89.5)?滋V and I50 measured (-3.30±0.36) log cd·s·m-2. Conclusion The electrophysiological characteristics of the rat are different from humans. Thus proper stimulation intensities must be adopted when using b-wave luminance response functions to evaluate the retina of the rat.
[Key words] eletroretinogram; SD rat; intensity response function
视网膜电图(electroretinogram,ERG)是视网膜电活动的总体反映,其中b波振幅最有代表性。由于b波的衡量标准属于一维测量,其正常范围与异常范围重叠程度较大,使其在临床和科研上的应用受到了很大限制。近年来电生理工作者对b波的光强—反应曲线及方程产生了浓厚兴趣,研究发现该方程中的相关参数可提供有关视网膜功能的更丰富信息,为临床应用提供了新的诊断指标。本研究采用Naka-Rushton方法分析了正常SD大鼠暗适应ERG b波光强—反应曲线,为今后的实验研究提供参考。
1 材料与方法
1.1 动物 清洁级成年SD大鼠12只(温州医学院实验动物中心提供,雌雄不限),体重为250~300 g。外眼和检眼镜检查屈光间质清晰,眼底无异常。予以12 h明-暗交替光照,不限食水,在室温18~23℃条件下饲养。
1.2 ERG检查方法 实验前大鼠暗适应12 h;在暗红光条件下用0.5%复方托吡卡胺滴眼液散瞳,0.5%丁卡因角膜表面麻醉;用速眠新Ⅱ注射液(0.8 ml/kg)肌肉麻醉,全麻后使其俯卧于立式支架的平台上,用特制支架进行固定,棉签揩干结膜囊。自制电极,其中记录电极由细银丝制成,分环和柄两部分,环部与角膜充分接触;参考电极和接地电极用不锈钢针制成,参考电极置于同侧颊部皮下,接地电极位于尾部皮下。
1.3 刺激条件 使用Ganzfeld Q400刺激器(ROLAND,德国),闪光强度分为7档,分别为-5.0、-4.5、-4.0、-3.5、-3.0、-2.5和-2.0 log cd·s·m-2(0.00003,0.0001,0.0003,0.001,0.003,0.01和0.03 cd·s·m-2),前5档强度通过人工放置自制中性滤光片获得(滤光片粘贴于刺激光源前)。白色闪光的色温为7000 K,实验时予以不同光强的光刺激,刺激间隔为1 min,通频带设为1~100 Hz,分析时间250 ms。
1.4 统计学方法 b波幅值用x±s表示,采用Naka-Rushton方法分析SD大鼠暗适应ERG b波光强—振幅曲线,计算出Rmax(b波饱和幅值)和I50(产生1/2饱和幅值时对应的刺激强度)。
2 结果
本实验利用一组强度从弱到强逐渐递增的闪光刺激SD大鼠暗适应视网膜,记录到ERG为一单纯的b波,波形光滑,其阈强度为-4.5 log cd·s·m-2,随着光强的增大,b波反应幅值逐渐增大,当刺激强度为-2.5 log cd·s·m-2时幅值趋向饱和(见表1,图1)。b波的这种光强和振幅间的关系可用一个光强—振幅曲线来表示,该曲线呈“S”形,通过Naka-Rushton方法分析,可以得到两个重要参数Rmax和I50,本实验中Rmax为(721.2±89.5)μV,I50为(-3.30±0.36)log cd·s·m-2(见图2)。
[1] [2] 下一页 |