兔肾体外灌注细胞脱落规律及其形态学改变

Shedding Law of Cells in the Perfused Rabbit Kidney

目的观察室温下21～25℃DMEM、血浆和血液+DMEM有氧灌注对离体兔肾的形态学影响,探讨离体肾在体外灌注情况下的细胞脱落规律。方法50只大耳白兔随机分为正常对照(5只)、DMEM组(15只)、血浆组(15只)、血液+DMEM组(15只),实验组观察时相点为灌注后1、3、6h。各组离体肾分别用相应的灌注液进行有氧灌注,通过大体观察、含水率、病理切片、脱落细胞记数、原位标记法(TerminaldeoxynucleotidyltransferaseTdTmediatedbiotindUTPnickendlabeling,TUNEL)凋亡染色进行灌注肾脏的细胞脱落规律分析。结果各组灌注后肾脏外观明显水肿,正常肾脏含水率为786%,灌注后肾脏含水率显著升高,DMEM组肾脏灌注6h后含水率为848%。光镜下见肾脏间质水肿、肾小球肾小管上皮细胞脱落、细胞核崩解、细胞密度减小。TUNEL染色正常对照凋亡细胞率为23%,DMEM组灌注1h细胞凋亡率即达159%。灌注液中脱落细胞数随灌注时间进行性增加。结论室温下离体肾脏有氧灌注后有较严重的肾脏上皮细胞脱落现象,凋亡和坏死均参与了其发生机制。

Objective To observe the morphologic changes of perfused kidney of rabbit and explore the law of shedding cells after kidney was perfused in room temperature(21-25)℃. Methods Fifty rabbits were divided into normal group, DMEM group, plasm group and blood+DMEM group. The kidney was perfused with different perfusion solutions respectively and observed at the designed time points of 1,3 and 6 hours post perfusion. The law of cell shedding was tested with water bearing rate, shedding cell counting, pathological observation, TUNEL (Terminal deoxynucleotidyl transferase TdT mediated biotin dUTP nick end labeling) staining. Results\ In all experimental groups, interstitial edema and renal tubular epithelial cell shedding were found in kidney cortex, especially in DMEM group. The water bearing rate of normal group was 78 6%,and it increased in experimental groups significantly. When perfused for 6 hours, the water bearing rate of the kidney in DMEM group was 84 8%. The ratio of apoptotic cells in normal group was 2 3%, and this ratio increased in all experimental groups. In DMEM group, the cell apoptosis ratio was 15 9% when kidney was perfused for only 1 hour. The number of the shedding cells increased with long perfusion. Conclusion\ Tubular epithelial cells shedding occurred after kidney was perfused. Apoptosis and necrosis were both involved in this injury.