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目的检测人脑组织库中保存的脑组织蛋白质、DNA及RNA质量,分析影响其质量的主要因素。方法选取中国医学科学院北京协和医学院人脑组织库保存的100例全脑组织样本,提取蛋白质、DNA及RNA,测定其浓度及吸光度,电泳检测完整性,用Western blotting及Rael-time PCR分别检测。根据不同年龄、死亡原因、取材延迟时间以及样本保存时间进行分组,统计学分析影响脑组织质量的因素。结果人脑组织库中大部分脑组织的DNA、RNA和蛋白质质量保存完好,不同年龄、死亡原因对其质量无显著影响。取材延迟时间超过24h者,其蛋白质浓度及RNA浓度及质量有显著下降。样本保存时间对蛋白质及DNA质量无显著影响,但保存1年以上可能对RNA浓度有影响。结论进行RNA相关分析的人脑组织标本取材延迟时间最好控制在8 h之内,-80℃环境下保存1年之内为佳。而DNA和蛋白质相关分析的时间可以适当延长。
Objective To detect the protein, DNA and RNA of brain tissue preserved in human brain tissue bank, and to analyze the main factors affecting its quality. Methods 100 whole brain tissue samples from the human brain tissue bank of Chinese Academy of Medical Sciences and Peking Union Medical College were collected for protein, DNA and RNA extraction. The concentration and absorbance were determined. The integrity of the whole brain was detected by electrophoresis. The protein expression was determined by Western blotting and Rael-time PCR . According to different age, cause of death, drawing delay time and sample preservation time grouping, statistical analysis of factors affecting brain tissue quality. Results The DNA, RNA and protein quality of most brain tissues in human brain tissue were well preserved. There were no significant effects on the quality of DNA at different ages and deaths. When the material was taken for more than 24 hours, its protein concentration and RNA concentration and quality were significantly decreased. Sample storage time had no significant effect on protein and DNA quality, but preservation for more than 1 year may have an impact on RNA concentration. Conclusion The RNA-related analysis of human brain tissue samples taken within the best control of the delay within 8 h, -80 ℃ environment within 1 year of preservation is better. DNA and protein-related analysis of the time can be appropriately extended.