The Autophagy Level Changes Of Rat Lung Transplantation For Pulmonary Ischemia Reperfusion

Objective:Experiment through the establishment of orthotopic lung transplantation model in rats, using the experimental zoology and molecular biology technology, expounds the autophagy level changes of rat lung transplantation for pulmonary perfusion,cold ischemia and reperfusion, and regulates the level of autophagy to reduce ischemia-reperfusion injury to provide theoretical basis for lung transplantation.Methods:1. Set up the group and establish the model: SPF male SD rats, a total of 60, the weight is between 300 ~ 350 g. Randomly selected 12 as control group, only pure perfusion cold ischemia processing, another 48 were divided into supply and receptors, receptor slightly larger than the donor, for difference of weight between, receptor does not exceed 20 g, on the left lung transplant. This experiment was divided into five groups:(1) CON group, cold ischemia preservation 0 h after perfusion group;(2) ISC6 group, cold ischemia preservation 6 h after perfusion group;(3) REP2 group, cold ischemia preservation 6 h after perfusion, reperfusion 2 h after transplantation group;(4) REP6 group, cold ischemia preservation 6 h after perfusion, reperfusion 6 h after transplantation group;(5) REP12 group, cold ischemia preservation 6 h after perfusion, reperfusion 12 h after transplantation group. There are 6 lung specimen in each group.2. Molecular Biological Detection: Immunofluorescence double dye LC3, LAMP- 2, Autophagic flux detection, Western blot detection LC3- II/LC3-1.Results:1. Through improved lung transplantation technique a kind of rat orthotopic left lung transplantation model was established successfully.2. By immunofluorescence double dye LC3, LAMP-2, and found that: Autophagic flux of group CON was the lowest, autophagic flux of group ISC6 increased slightly, autophagic flux of group REP2, REP6 increased significantly and the highest was group REP6. But autophagic flux of group REP12 drceased.3.By Western blot detection LC3 protein expression found: CON group of LC3 protein expression was lowest,ISC6 group of LC3 protein expression was a bit higher, REP2, REP6 group of LC3 protein expression rise significantly, the highest LC3 protein expression was group REP6, REP12 group of LC3 protein expression decreased.Conclusion:Rat lung transplantation for pulmonary perfusion, cold ischemia autophagy level began to increase after 6h and increased obviously after reperfusion 2h. Reperfusion autophagy level reaches its peak after 6 hours, as the reperfusion time extend the autophagy level will weaken,the autophagy level of 12 h was less than the autophagy level of 6 h.