不同温度对人体髂静脉保存效果的影响  

Effects of different temperatures on preservation of human iliac vein

作　　者：任章勇 吕少诚[1] 王芳菲 赵昕[1] 郎韧[1] 贺强[1] REN Zhangyong;LYU Shaocheng;WANG Fangfei;ZHAO Xin;LANG Ren;HE Qiang(Department of Hepatobiliary Surgery,Beijing Chaoyang Hospital,Capital Medical University,Beijing 100020,China)

机构地区：[1]首都医科大学附属北京朝阳医院肝胆外科,北京100020

出　　处：《解放军医学院学报》2022年第12期1259-1265,共7页Academic Journal of Chinese PLA Medical School

基　　金：首都医科大学附属北京朝阳医院临床研究孵化项目(CYFH202214)。

摘　　要：背景低温冷藏保存(cold storage,CS)是目前临床医疗实践中常用的离体血管组织短期保存方式,但其有效保存时间及其对离体血管结构及功能的影响尚缺乏系统性的研究。目的探索低温冷藏与深低温冷冻对人体髂静脉保存效果的影响。方法来自12例器官捐献者的24条髂静脉作为研究对象,将其修剪为长约1.5 cm的血管段并随机分为低温冷藏保存组和深低温冷冻保存(frozen storage,FS)组,每个组240个静脉段。分别于保存1 d、3 d、5 d、1周、2周及4周时取出血管,通过MTT比色法及TUNEL荧光染色评估血管细胞存活情况和凋亡情况,通过极限应力试验评估血管机械性能,通过HE染色、Masson染色和EVG染色评估血管内皮细胞、平滑肌细胞、胶原纤维及弹性纤维的变化。结果随着保存时间的延长,两组血管的凋亡细胞均逐渐增多,细胞活性逐渐下降,抗张力性能逐渐减弱;在保存5 d内冷藏组及冷冻组均未出现明显的凋亡细胞,但随着保存时间延长,两组凋亡细胞均逐渐增多,在1周(6.09%±0.90%vs 3.96%±0.69%,P=0.001)、2周(34.58%±3.40%vs 17.45%±1.74%,P<0.001)及4周(74.90%±5.68%vs 31.23%±1.52%,P<0.001)时冷藏组凋亡指数均高于冷冻组;在细胞活性方面,在保存1 d[(2.27±0.22)OD/m vs(1.64±0.30)OD/m,P=0.002]和3 d[(2.11±0.14)OD/m vs(1.59±0.31)OD/m,P=0.001]时冷藏组细胞活性高于冷冻组,3 d~2周时两组细胞活性相当,2周[(0.83±0.24)OD/m vs(1.29±0.15)OD/m,P=0.001]及4周[(0.48±0.27)OD/m vs(1.15±0.27)OD/m,P<0.001]后冷藏组细胞活性低于冷冻组;在抗张力方面,1 d[(11.98±0.55)N vs(10.96±0.56)N,P=0.004]、3 d[(11.88±0.40)N vs(10.89±0.60)N,P=0.002]和5 d[(11.72±0.71)N vs(10.97±0.55)N,P=0.004]时,冷藏组抗张力性能强于冷冻组;1~2周之间两组抗张力效果相当,4周后冷藏组的抗张力性能弱于冷冻组[(10.11±0.48)N vs(10.73±0.62)N,P=0.042]。在血管管壁结构方面,冷藏保存2周后血管内皮细胞可见破坏脱落,Background Cold storage scheme is a commonly used short-term preservation method of isolated vascular tissue in clinical practice at present,but its effective preservation time and its impact on the structure and function of isolated vascular tissue are still lack of systematic research.Objective To explore the advantages and disadvantages of cold storage and frozen storage for the preservation of human iliac vein.Methods Human iliac veins were randomly divided into Cold Storage(CS)group and Frozen Storage(FS)group,and they were stored for 1 d,3 d,5 d,1 week,2 weeks and 4 weeks.The mechanical properties of blood vessels were evaluated by ultimate stress test,and the survival and apoptosis of vascular cells were evaluated by MTT colorimetry and TUNEL fluorescence staining.The changes of vascular endothelial cells,smooth muscle cells,collagen fibers and elastic fibers were evaluated by HE staining,Masson staining and EVG staining.Results With the prolongation of storage time,the apoptotic cells of blood vessels in both groups gradually increased,the cell viability gradually decreased,and the tensile properties gradually weakened.No significant apoptotic cells appeared in the CS group and FS group within 5 days of storage,but with the prolongation of storage time,the apoptotic cells in both groups gradually increased,and the apoptotic indices in the CS group were higher than those in the FS group at 1 week(6.09%±0.90%vs 3.96%±0.69%,P=0.001),2 weeks(34.58%±3.40%vs 7.45%±1.74%,P<0.001),and 4 weeks(74.90%±5.68%vs 31.23%±1.52%,P<0.001).In terms of cell viability,the apoptotic indices in the CS group were higher than those in the FS group at 1 day([2.27±0.22]OD/m vs[1.64±0.30]OD/m,P=0.002)and 3 days([2.11±0.14]OD/m vs[1.59±0.31]OD/m,P=0.001).Cell viability was comparable between the two groups from 5 days to 2 weeks,and it was lower in the CS group than that of FS group after 2 weeks([0.83±0.24]OD/m vs[1.29±0.15]OD/m,P=0.001)and 4 weeks([0.48±0.27]OD/m vs[1.15±0.27]OD/m,P<0.001).At day 1([11.98±0.55]N vs[1

关 键 词：同种异体血管 血管移植物 血管保存 深低温冷冻 低温冷藏 

分 类 号：R446.11[医药卫生—诊断学]