Chinese Journal of Tissue Engineering Research ›› 2011, Vol. 15 ›› Issue (5): 813-817.doi: 10.3969/j.issn.1673-8225.2011.05.013

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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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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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