| Objective Research the Ex Vivo technology of oxygenated blood at room temperature continuous perfusion through the pulmonary artery to prevent ischemic-reperfusion injury of donor lung without using ventilator,and to observe the relation of alveolar apoptosis with ischemic-reperfusion injury of donor lung.Provide experimental basis for long time preservation of lung and long distance transportation.Methods Adoption 24 healthy purebred piglets from Bama 24, were randomly divided into two groups,namely:the experimental group (oxygenated blood at room temperature continuous perfusion through the pulmonary artery 8h to protect donor lung without using ventilator,n=12)and control group (Celsior solution by pulmonary artery perfusion at 4℃ cryopreservation 8h for lung protection, n=12).Then the left lung was transplanted and with mechanical ventilation for 24h.Pulmonary function indexes,including lung ventilation resistance(LR),blood gas index(the left inferior pulmonary vein blood PO2(LIPV-PO2)and lung water content, were evaluated.To classify pulmonary edema grading and measure the death rate of lung cell by HE staining.Through the TUNEL method combined with transmission electron microscope to observe the level of alveolar cell apoptosis, apoptosis of alveolar cell types and the trend of apoptosis.Result 1. Pulmonary function test:Comparison at the same time after surgery, the lung water content, LR of experimental group was significantly lower than that of the control group, LIPV-PCO2 was significantly higher than that of control group (P< 0.05); after operation 24h in the experimental group lung water content, LR was significantly less than 12 hours after surgery, LIPV-PCO2 was significantly higher than that of postoperative 12 h (P< 0.05). 2.HE staining was observed in experimental group at each time point, lung tissue injury was less than the control group; control group, the lung tissue injury after operation 24h is most serious,70%-80% of alveolar appeared different degree edema or hemorrhage, even large areas of hemorrhage and necrosis. In the experimental group, after surgery 12h was the most serious, 20-30% alveolar cavity appeared edema fluid, local alveolar cavity and pulmonary interstitial hemorrhage, and after 12h, there was a trend of improvement. Alveolar cell death rate:Comparison at the same time after surgery, the experimental group was significantly lower than that of the control group (P< 0.05); After surgery 24h,the alveolar cell death rate of control group increased gradually,24h after operation to reach 32.7±4.6%; after surgery in the experimental group alveolar cell death in postoperative 12h reached the highest 15.8±2.1%.3.Detection of Tunel:Comparison at the same time after surgery,the alveolar cells apoptosis rate of experimental group was significantly lower than that of the control group (P< 0.05). After operation in the experimental group at 6h highest 8.2±1.5%, postoperative 24h alveolar cell apoptosis rate was significantly lower than that of postoperative 6h (P<0.05); postoperative 24h alveolar apoptosis rate and lung water content, LR was positively significantly correlated (r=0.890,0.918, P< 0.05), and LIPV-PCO2 was significantly negatively related (r=-0.784, P< 0.05); 4. Transmission electron microscope:Apoptosis of alveolar cells was mainly AEC II, about accounted for 80%. In the experimental group, the level of apoptosis of alveolar cells was lower than that of control group, and the rising trend of apoptosis in the experimental roup was earlier than that of the control group.Conclusion 1.Oxygenated blood at room temperature continuous pulmonary through artery perfusion with donor lung preservation can effectively reduce the LIRI, make improvements in lung function after lung transplantation. The donor lung preservation effect is better than the way of "static cryopreservation"; 2.1n the early stage of lung transplantation (less than 24h), there was a significant correlation between the apoptosis rate and LIRI, the higher the level of apoptosis, the more serious the degree of LIRI damage. And the main types of alveolar cell apoptosis were AECIL; 3.The method of perfusion and preservation of oxygenated blood at room temperature may reduce the level of alveolar cell apoptosis,then reduce the LIRI, and play a protective role for donor lung. |
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