摘要
目的 探讨离体脊髓超薄切片的生物活性以及具有生物活性脊髓片的生存时间窗.方法 选取Wister大鼠（6～7周）24只,用微型切片机制备胸段脊髓横切片（厚400 μm）.脊髓片在自制双聚苯乙烯孵化箱内采用5％ 02/5％ C02持续灌注的克-林氏液（36℃）中进行孵化,采用正电子放射自显影法（dPAT）,测量脊髓组织区域葡萄糖代谢率（MRglc）变化为参考指标,考察离体脊髓片的生物活性及具有生物活性脊髓片的生存时间窗.结果 在离体后6h内脊髓组织氟代脱氧葡萄糖（[18 F]FDG）吸收曲线具有很好的线性拟合（r=0.99）,[18F] FDG的相对吸收值即MRglc保持恒定（K =7.49±1.27）,缺氧、河豚毒素、高K＋ （50 mmol/L）溶液等能明确诱发出与脊髓神经元的功能改变相关的MRglc变化,K值分别为：-0.09（45 min）、6.17、11.17,表明离体后6h内脊髓片具有的神经活动依赖性代谢.结论 脊髓片在离体后6h内仍保存了脊髓神经元的生理功能以及神经活动依赖性代谢,脊髓横切片（厚400 μm）是进行离体脊髓保护及电生理等实验可利用模型.
Objective We attempted to measure the bioactivity of sliced spinal cord in vitro and the survival time window of bioactive spinal cord slices,which laid the scientific foundation for using disembodied spinal cord slice for further study,like spinal cord protection and electrophysiological experiment and so on.Methods Thoracic spinal cord from 24 Wister rats （6-7 weeks old） were cut into 400 μm slices by microslicer.The spinal cord slices were transferred into double polystyrene chambers and incubated in Krebs-Ringer solution at 36 ℃,bubbled with 5% O2/5% CO2 gas.The changes of regional metabolic rate of glucose （MRglc） were measured by dynamic positron autoradiography technique （dPAT） as an index for bioactivity of spinal cord slices and the survival time window of bioactive spinal cord slices.Results The Uptake curves of [18F]-fluoro-deoxy-D-glucose （FDG） in the spinal cord slices were well fitted by straight line （r =0.99） within 6 hours after the slicing of the spinal cord.The relative uptake value of [18F] FDG,MRglc,remained constant （K =7.49 ± 1.27）.The MRglc associated with spinal neurons＆#39; function is affected by hypoxia,tetrodotoxin and high K （50 mmol/L） solution,with K value-0.09 （45 min）,6.17,11.17,respectively,indicating the spinal cord slices retained an activity-dependent metabolism within 6 hours in vitro.Conclusion Spinal cord slices retained spinal neurons physiology function and an activity-dependent metabolism within 6 hours in vitro.Spinal cord slice （400 μm） is a suitable model for protection and electrophysiological experiment of disembodied spinal cord.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2016年第5期1301-1304,共4页
Chinese Journal of Experimental Surgery
基金
广东省自然科学基金（S201310013837）
关键词
脊髓
氟代脱氧葡萄糖
大鼠
葡萄糖代谢
Spinal cord
[18F]-fluoro-deoxy-D-glucose
Rat
Glucose metabolism
