眼内灌注液对鼠视网膜神经上皮层细胞活力影响的体外实验研究  

In vitro experimental study of the effects of different introcular irrigating solutions on the viability of rat retinal neuron cells

作　　者：姚进[1] 黎晓新[1] 万有[2] 薛冰[2] 周慧芳[2] 

机构地区：[1]北京大学人民医院眼科中心,100044 [2]北京大学神经科学研究所

出　　处：《中华眼科杂志》2004年第7期453-458,共6页Chinese Journal of Ophthalmology

摘　　要：目的 比较不同眼内灌注液对视网膜神经上皮层细胞活力及抗氧化作用的影响,以寻找理想的眼内灌注液。方法原代培养新生大鼠视网膜神经上皮层细胞分为A、B、c、D、E组,其中A组为对照组,为含血清DMEM/F12培养液,其余4组分别用BSS plus、BSS plus+药物(妥布霉素16000 U/L、地塞米松16mg/L和盐酸肾上腺素1mg/L)、自制灌注液和乳酸林格液+药物4种眼内灌注液,分别孵育细胞4、12、24、48 h后,检测各组细胞线粒体琥珀酸脱氢酶活性、乳酸脱氢酶(LDH)释放率和细胞凋亡的阳性率;在不同灌注液中加入30μmol/LH2O2诱导细胞氧化损伤,4 h后检测各组细胞的脂质过氧化物分解产物丙二醛(MDA)的含量。结果各灌注液组细胞线粒体琥珀酸脱氢酶活性随时间延长呈下降趋势,LDH释放率和凋亡阳性率呈上升趋势(P<0.05),而同一时间点不同组对细胞活力影响由小到大为:B组、C组、D组和E组,其中C组和D组差异无显著意义(P>0.05)。在抗氧化作用方面,B组、C组和D组的细胞内MDA含量明显少于E组(P<0.01),且3组之间差异无显著意义(P>0.05)。结论 自制灌注液在乳酸林格液中添加了碳酸氢盐、葡萄糖、氧化型谷胱甘肽及药物妥布霉素、盐酸肾上腺素、地塞米松后,除具有与BSS plus液相似的抗氧化能力和缓冲能力外,还提供了更多的能量来源和抗感染、抗?Objective To observe the effects of different introcular irrigating solutions on the viability of retinal neuron cells , their antioxidant ability, and to find the ideal introcular irrigating solution. Methods Retinal neuron cells from Sprague Dawley rats ( postnatal 1-3 days) were cultured in the 4 different introcular irrigating solutions which were BSS plus, BSS plus + drugs(tobramycinl. 6 ten thousand unit /L、desamethasone 16 mg/L and adnephrin 1 mg/L) , new irrigating solution and lactated Ringer's solution + drugs. Cell viability was determinated by MTT [ 3 , (4, 5-Dimethylthiazol-2-yl) 2, 5-diphenyl-tetrazolium bromide ] assay, LDH release was tested by LDH assay and apoptotic cell death was evaluated by the TUNEL(TdT-mediated dUTP nick-end labeling) assay after retinal neuron cells were cultured in the different introcular irrigating solutions for 4,12,24,48 hours. MDA ( malondialdehyde) production was tested after the cells exposed to 30 μmol/L H2O2 for 4 hours. All data was analyzed in Dunnett's ANOVA way. Results The cell viability was decreased and the rate of LDH released and cell apoptosis was increased. These effects were shown to be time-dependent for the same solution. At the same time point, the effects on cellular survive was variable in deferent introcular irrigating solutions, BSS plus is the lowest, BSS plus + drugs and modified irrigating solution are midst, and lactated Ringer's solution + drugs is the highest. The result of MDA assay showed that both BSS plus and modified irrigating solution can inhibit the oxidative damage induced by H2O2. Conculsion Modified introcular irrigating solution may be an better introcular irrigating solution according to its little effect on cell viability , its antioxidant function and lower cost ( Chin J Ophthalmol, 2004, 40:453458

关 键 词：眼内灌注液 鼠 视网膜神经上皮层 细胞活力 实验 细胞凋亡 琥珀酸脱氢酶活性 

分 类 号：R774.1[医药卫生—眼科]