中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (5): 813-817.doi: 10.3969/j.issn.1673-8225.2011.05.013

• 肾移植 kidney transplantation • 上一篇    下一篇

迟发缺血预处理低温保存肾移植供体中血红素加氧酶1的表达

李  飞1，2，周忠兴2，陆曙炎2，朱瑞霞1，3，马玲娣1，蔡雷铭4   

  1. 南京医科大学附属常州第二人民医院，1中心实验室，2泌尿外科，3骨科，4病理科， 江苏省常州市，213000
  • 收稿日期:2010-08-09 修回日期:2010-11-10 出版日期:2011-01-29 发布日期:2011-01-29
  • 通讯作者: 陆曙炎，主任医师，教授，硕士生导师，南京医科大学附属常州第二人民医院泌尿外科，江苏省常州市 213000 lf19841107@foxmail.com
  • 作者简介:李飞★，男，1984年生，山西省长治市人，汉族，南京医科大学在读硕士，主要从事肾脏移植方面的研究。 281411053@qq.com

Heme oxygenase-1 expression in cryopreserved kidney grafts during delayed ischemia preconditioning

Li Fei1, 2, Zhou Zhong-xing2, Lu Shu-yan2, Zhu Rui-xia1, 3, Ma Ling-di1, Cai Lei-ming4   

  1. 1Central Laboratory, 2Department Urology, 3Department of Orthopaedics, 4Department of Pathology, Changzhou Second People’s Hospital Affiliated to Nanjing Medical University, Changzhou  213000, Jiangsu Province, China
  • Received:2010-08-09 Revised:2010-11-10 Online:2011-01-29 Published:2011-01-29
  • Contact: Lu Shu-yan, Chief physician, Professor, Master’s supervisor, Department Urology, Changzhou Second People’s Hospital Affiliated to Nanjing Medical University, Changzhou 213000, Jiangsu Province, China lf19841107@foxmail.com
  • About author:Li Fei★, Studying for master’s degree, 1Central Laboratory, 2Department Urology, Changzhou Second People’s Hospital Affiliated to Nanjing Medical University, Changzhou 213000, Jiangsu Province, China 281411053@qq.com

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摘要：

背景：缺血预适应延迟反应通过诱导保护性蛋白增强组织对缺血再灌注损伤的耐受能力；血红素加氧酶1参与缺血预适应延迟保护作用。迟发缺血预处理对低温保存肾脏的作用及血红素加氧酶1是否参与其中尚不清楚。
目的：观察缺血预处理诱导血红素加氧酶1的迟发缺血预处理反应对低温保存肾脏移植供体的作用。
方法：雄性SD大鼠随机分入5组：空白对照组、低温保存组、缺血预处理组、缺血+低温组(n=12)；缺血+给药+低温组。各组大鼠均行右肾切除，预处理或假手术操作处理后24 h采用大鼠肾脏非循环离体灌注模型获取肾脏，分别于保存24，48，72 h取样。缺血+给药+低温组除上述处理外，还于原位低温灌注术前1 h接受1次血红素加氧化酶1抑制剂锡原卟啉腹腔注射。低温保存肾脏于各保存终点留取保存液，测定pH值和乳酸脱氢酶含量；切取1/2肾脏按照光镜要求制备标本送检；剩余1/2肾脏用于免疫印迹法测定血红素加氧酶1表达，比色法测定皮质Na-K-ATP酶活性、丙二醛和还原型谷胱甘肽含量；未保存肾脏仅通过免疫印迹法测定血红素加氧酶1的基础表达情况。
结果与结论：迟发缺血预处理诱导了肾组织血红素加氧酶1的表达，与单纯低温保存组相比保存24，48 h后，缺血+低温组保存液pH值、乳酸脱氢酶活性降低；肾脏组织Na-K-ATP酶活性、谷胱甘肽含量增加，丙二醛含量降低；同时点预处理组肾组织光镜形态学改变稍好于单纯低温保存组。给予血红素加氧酶1抑制剂后，这种保护作用消失。提示，迟发缺血预处理延长了肾脏低温保存时限，这可能与诱导血红素加氧酶1，增加组织抗氧化能力，减轻低温保存氧应激有关。

关键词: 血红素加氧酶1, 肾脏移植, 缺血预适应, 低温保存, 应激

Abstract:

BACKGROUND: Delayed response of ischemia preconditioning alleviates organs ischemia reperfusion injury through induction of protective proteins. Heme oxygenase-1 (HO-1) is involved in delayed protection of ischemic preconditioning. However, the effect of delayed ischemia preconditioning on cryopreserved kidney grafts and whether HO-1 is involved in this process remains unknown.
OBJECTIVE: To explore the effect of delayed ischemia preconditioning response of HO-1 induced by ischemia preconditioning on cryopreserved kidney grafts.
METHODS: Male Sprague-Dawley rats were randomly divided into 5 groups: control-sham group, cryopreservation group, ischemia preconditioning group, ischemia + cryopreservation group (n=12), ischemia + administration + cryopreservation group. Rats in all groups underwent resection of right kidney. At 24 hours after pretreatment or sham operation, kidneys were obtained from live rats via the method of renal non-circulation isolated and reperfused rat model, then the samples was collected after 24 hours, 48 hours, 72 hours hypothermal preservation. Ischemia + administration + cryopreservation group got additional procedure that was a single injection of HO-1 inhibitor Sn-protoporphyrin intraperitoneal injection at one hour before situ hypothermal perfusion surgery. At each preservation end point, the preservation solution was collected for detection of pH values and lactate dehydrogenase (LDH) content. Half of the preserved kidney was obtained and prepared for detection according to light microscope requirements, while the remaining kidneys were used to detect HO-1 expression by immunoblotting; cortical Na-K-ATP activity, and contents of malonaldehyde (MDA) and reduced glutathione hormone (GSH) were detected by colorimetric method. Kidneys without cryopreservation were used to detect basal expression of HO-1 by western blotting.
RESULTS and CONCLUSION: Delayed ischemia preconditioning induced HO-1 expression in renal tissue. Compared with cryopreservation group, preservation solution pH values and LDH activity in ischemia + cryopreservation group were reduced after 24 hours, 28 hours. The contents of Na-K-ATP activity and GSH were increased and the content of MDA was decreased in renal tissue. Meanwhile, the renal morphological changes of light microscopy in ischemia preconditioning group slightly better than that of cryopreservation group at the same time point. However, this protective effect disappeared after the injection of HO-1 activity inhibitor. It is indicated that the elongation of renal hypothermal preservation duration may be associated to the induction of HO-1, the increase of tissue antioxidant ability and the reduction of hypothermia caused oxidative stress.

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