肾脏脱细胞基质结构功能重建的初步研究  

Preliminary Study about Reconstruction of the Structure and Function of Decellular Matrix Kidney

作　　者：王磊[1] 许宏[2] 王珏[3] 雷永华[1] 付强[1] 邱建新[1] 保庭毅[1] 

机构地区：[1]第四军医大学唐都医院泌尿外科,陕西西安710038 [2]解放军161医院肾脏内科,湖北武汉430000 [3]第四军医大学唐都医院感染科,陕西西安710038

出　　处：《现代生物医学进展》2013年第6期1038-1044,共7页Progress in Modern Biomedicine

摘　　要：目的:本实验利用多种脱细胞剂通过半自动体外循环灌注洗脱肾脏细胞,摸索出最佳脱细胞剂种类及脱细胞方案。方法:以健康兔肾为研究对象,采用半自动体外循环灌注法,在10-15 ml/min流速及不同浓度、温度条件下经肾动脉灌注各种脱细胞剂,观察肾脏灌注过程中变化,灌注结束解剖肾脏,大体观察后行苏木精-伊红(HE)染色及组织形态学观察,重点观察肾小球、肾小管细胞结构的变化。结果:比较理想的肾脏脱细胞方案是1%Ⅳ型胶原酶-37℃,20 min,该酶能在相对较短的时间内洗脱部分肾实质细胞而对肾脏基质未造成明显损伤。该方案灌注时间短,能减少肾脏热缺血时间,有助于肾脏再生。其次是0.5%胰蛋白酶-25℃,2h,该方案能使肾小球细胞比较彻底的脱出,只留基质部分,对肾小管作用较小,但该方案用时较长,已达到肾脏体外耐受缺血的极限,理论上肾脏灌注后很可能会因缺血缺氧时间过长而坏死,故不推荐使用。洗脱效果最强的脱细胞剂是SDS,但其对肾脏实质结构造成不可恢复的破坏,尤其是引起基底膜断裂不利于肾实质细胞再生。其他脱细胞方案均不理想。结论:本研究证实通过选择适当的脱细胞剂,配合适合的温度、浓度、灌注压等条件,是可以实现肾脏部分脱细胞基质的,为下一步研究衰竭肾脏的部分脱细胞基质甚至是在体灌注乃至脱细胞后肾脏结构功能的重建提供了实验依据。Objective： This experiment is only a preliminary study to explore the optimal dose of decellular agents and perfusion programs by a semi-automatic renal perfusion to elute parts of cells and matrix of renal. Methods： Different decellular agents in 10-15 ml/min velocity and temperature conditions were perfused into healthy rabbit kidneys through renal artery by semi-automatic pump perfusion system. Changes in the course of renal perfusion should be observed. Kidneys were dissected and underwent hematoxylin-eosin stain to observe the general histological examination, focusing on glomerular and tubular cell structure changes. Results： The best decellular agent and perfusion scheme is 1% collagenase 1V at 37 ℃ for 20min, the enzyme can elute renal parenchyma cells within a relatively little time without side injury to renal matrix, and it can reduce the warm ischemia time of renal, which contributes to kidney regeneration. The second best decellular agent and perfusion scheme is 0.5 % trypsin at 25 ℃ for 2h, which can elute more glomerular cells, leave only the matrix part and have little effect on renal tubular, but this perfusion scheme with a longer time, has reached the resistance limit of kidney in vitro to ischemia. Thus, it is not recommended. The most powerful decellular agent is SDS, but the renal caused irreversible damage, especially the damage of basement membrane is not conducive to regeneration of renal. Other perfusion schemes are not ideal. Conclusion： By selecting the appropriate decellular agent, with a suitable temperature, concentration, perfusion pressure and other conditions, parts of renal cells and matrix can be eluted, which will provide an experimental basis for the reconstruction of the structure and function of nonfunctioning kidney.

关 键 词：脱细胞 组织工程 肾功能衰竭 兔 

分 类 号：Q95-3[生物学—动物学] R692.5[医药卫生—泌尿科学]