冷缺血时间与胆道闭锁肝组织RNA质量的相关性分析  

Correlation between cold ischemia time and RNA quality in hepatic tissue of biliary atresia

作　　者：马婷婷[1] 曹磊 王建喜 毕森盛 段可然 冯湛 毕博文 王政禄[1] 郑虹[1,2] MA Ting-ting;CAO Lei;WANG Jian-xi;BI Sen-sheng;DUAN Ke-ran;FENG Zhan;BI Bo-wen;WANG Zheng-lu;ZHENG Hong(Biobank,Tianjin First Central Hospital,Tianjin 300192,China;Oriental Organ Transplant Center/Key Laboratory of Transplantation,Chinese Academy of Medical Sciences,Tianjin First Central Hospital,Tianjin 300192,China)

机构地区：[1]天津市第一中心医院生物样本资源共享中心,300192 [2]天津市第一中心医院东方器官移植中心/中国医学科学院移植重点实验室,300192

出　　处：《中国医药生物技术》2022年第3期220-225,共6页Chinese Medicinal Biotechnology

基　　金：天津市第一中心医院春风项目(2019CF36);天津市卫生健康委员会科技项目(ZC20065)。

摘　　要：目的分析冷缺血时间与胆道闭锁肝组织样本中RNA浓度、纯度及完整性(RIN)的相关性,为获得高质量肝组织样本提供标准。方法选取天津市第一中心医院30例因胆道闭锁行肝移植术患者的肝组织,室温放置0、0.25、0.5、1、2、3、5和12 h后分别加入RNAlater,4℃过夜,提取总RNA,检测RNA浓度、纯度及RIN值,根据浓度计算出产量,运用单因素方差分析分析冷缺血时间与胆道闭锁肝组织样本中RNA产量、纯度及RIN值的相关性,典型相关分析分析不同冷缺血时间后组织样本RNA产量、纯度及RIN值之间相关性。结果组织样本冷缺血0、0.25、0.5、1、2、3、5和12 h后,每10mg组织样本的RNA产量分别是(9.123±0.530)、(9.220±0.606)、(9.000±0.862)、(9.067±0.498)、(9.027±0.523)、(9.143±0.627)、(9.113±0.446)和(9.193±0.492)μg(P=0.78)。RNA纯度的比值分别为1.914±0.065、1.896±0.060、1.904±0.053、1.907±0.057、1.896±0.057、1.888±0.050、1.908±0.060和1.899±0.059(P=0.74)。RIN值分别为9.077±0.104、8.777±0.138、8.650±0.187、8.197±0.188、7.963±0.227、7.597±0.301、4.433±0.272和3.747±0.236(P=0.000)。单因素方差分析显示冷缺血时间与RNA的产量和纯度均无相关性,与RIN值显著相关。典型相关分析显示不同冷缺血时间后组织样本RNA产量与纯度、产量与RIN值及纯度与RIN值之间均无相关性。结论冷缺血时间与胆道闭锁肝组织RNA质量具有显著相关性,缩短冷缺血时间可获得高质量RNA。Objective To analyze the correlation between cold ischemia time and RNA concentration, purity and RNA integrity number(RIN)in hepatic tissue samples of biliary atresia, so as to provide a standard for obtaining high-quality hepatic tissue samples.Methods Thirty patients with biliary atresia undergoing liver transplantation in the Tianjin First Central Hospital were selected.After removal of the hepatic tissue, RNAlater reagent was added at room temperature for 0, 0.25, 0.5, 1, 2, 3, 5 and 12 h. Total RNA was extracted, and the concentration, purity(A260/A280) and RIN were detected. The yield was calculated according to the concentration. One way ANOVA was used to analyze the correlation between cold ischemia time and RNA production, purity and RIN in hepatic tissue samples of biliary atresia. Canonical correlation was used to analyze the correlation between RNA production,purity and RIN in tissue samples after different cold ischemia time.Results The RNA yields(μg/10 mg) of the tissue samples after 0, 0.25, 0.5, 1, 2, 3, 5, and 12 h at room temperature were 9.123 ±0.530, 9.220 ± 0.606, 9.000 ± 0.862, 9.067 ± 0.498, 9.027 ± 0.523, 9.143 ± 0.627, 9.113 ± 0.446 and 9.193 ± 0.492, respectively(P = 0.78). The respective RNA purities(A260/A280) were 1.914 ± 0.065, 1.896 ± 0.060, 1.904 ± 0.053, 1.907 ± 0.057, 1.896 ± 0.057,1.888 ± 0.050, 1.908 ± 0.060 and 1.899 ± 0.059(P = 0.74). The RNA integrity number(RIN) were 9.077 ± 0.104, 8.777 ± 0.138,8.650 ± 0.187, 8.197 ± 0.188, 7.963 ± 0.227, 7.597 ± 0.301, 4.433 ± 0.272 and 3.747 ± 0.236(P = 0.000). The results from one way ANOVA showed that there existed no correlation of cold ischemia time with RNA yield and purity, but a significant correlation with RIN. The results from canonical correlation analysis showed that there was no correlation between RNA yield and purity, yield and RIN, purity and RIN after different cold ischemia time.Conclusion There is a significant correlation between cold ischemia time and RNA quality in liver tissue of biliary at

关 键 词：冷缺血时间 胆道闭锁 肝组织 RNA质量 RNA完整性 

分 类 号：R657.4[医药卫生—外科学]