The Protection Of UTI And GSH For Lungs In The Pulmonary Ischemical Reperfusion Under The Models Of Lung Transplantation

Objective:To investigate the protection of UTI and GHS for the lungs in the pulmonary ischemical reperfusion under the models of lung transplantation.Method:1. The animal: Selection 32 New Zealand adult white rabbits, each Weight 2.5-3.0Kg. (Provied by the animal experiments center of the Faculty of Medicine of Jiangsu University).2. Animal groups: The rabbits were devided into A,B,C&D, four groups randomly, 8 in each group. A group was the control group, B group was Ulinastatin group, C group was Glutathione group, while D group was Ulinastatin & Glutathione group.3. To fix the rabits to supine position after the intravenous anesthesia by 20% urethane (5ml/Kg) from the marginal ear vein, then to stablish intravenous access, maintain intravenous infusion by speed 10ml/(Kg. h). To heparinize (2mg/kg. To intubate by tracheotomy, control the breath by breathing machine: inhale oxygen concentration 21%, tidal volume 15ml/Kg, breathing frequency 40times/min. the time of inhale to exhale is 1:2,the positive end-expiratory pressure was 3cmH20. To monitor the pressure of system he right common carotid artery. To cut 3, 4, 5 cartilage since the left parasternal, to cut the whole floor to the chest wall to the axillary line along No. 2,5 left intercostal, to open chest, break the No. 3, 4, 5 costal bone at the axillary line, to expose the organs in the left chest, hemostasis carefully. To push and separate the pulmonary hilar organizations with Cotton Carriers, to isolate the left uper pulmonary artery, across a No. 10 line. To puncture the left pulmonary artery and intubate, pass the end to the left pulmonary artery crossover, ligate, the other end of the tube is fixed at the chest wall to prevent slippage. To ligate the left upper lobe organizations, cut out the left upper lobe of lung, to expose left lower lobe, to shear the left broad ligament of lung, to free organizations of the left hilum, to open the pericardium at the left side to expose connective part of the artery of the left lower lobe and the atrium pulmonale. To seam a pocket with sliding Line 5-0 in the left atrial wall, to puncture atrial wall, to intubate to the vena pulmonalis inferior sinistra, use the line to tie a knot, and fixthe tube, and fix the other end to the chest wall to prevent slippage. The pulmonary artery is connected with a sensor to the electrocardiogram monitor to monitor the pressure of the pulmonary artery. To block the lower left pulmonary artery, the lower left pulmonary vein, the lower left lung bronchus one by one. Cut down tidal volume to 10ml/Kg at the same time. After aperiod of time, to irrigate the 0℃lung protection solution from the tube which was intubated in the left uper pulmonary artery, and drainage the lung preservation solution from the tube which was intubated in the lower left pulmonary vein. Drive the pulmonary perfusion with the gravity. The vertical height is 60cm. The total perfusion liquid is 60 ml/kg. The perfusion liquid in A,B,C&D four groups are LPD liqiud, LPD liqiud containing UTI 20,000 U/Kg, LPD liqiud containing GSH 5mmol /L, and LPD liqiud containing UTI 20,000 U/Kg & GSH 5mmol /L. After the perfusion, to trap the left lung with double-thin plastic bag rapidly, to anti-pack it at the hilar department, to use a lateral wall clamp to block the plastic bag together with left pulmonery root. the Inner plastic bag is filled with preservation fluid which is the same as the protection solution. The outer plastic bag is filled with mixture of ice and water to maintain the temperature to 0℃, to keep the lower left lung fully immersed in the 0℃preservation solution, and time to 3 hours. To take away the preservation liquid and the plastic bag, open the lower left pulmonary vein, the lower left lung bronchus, the lower left pulmonary artery one by one, to recuperate the infuse of the left lung. To adjust the breathing machine to make the tidal volume 15ml/Kg, keep it breathing for 2 hours, obtain the sample at each time point. Kill the animals after the operations.Results:1. The comparison of the lung venous blood gas analysis and the pulmonary arterial pressure after the reinfusion: 5 minutes after reperfusion lung venous blood gas analysis shows: PO₂ in the UTI&GSH group and the control group was statistically significant(P<0.05). The others and the pulmonary artery pressure show no obvious differences, every group is statistically meaningless. 20 minutes after reperfusion, the comparison of PO₂ from lung venous blood gas analysis in control group UTI group, GSH group and UTI &GSH group shows P<0. 05, the pulmonary arterial pressure shows P<0.001, both have statistical significance. And the comparison between UTI&GSH group and the UTI group, GSH group also has statistical significance(P<0.05). 1 hour after the reinfusion the comparison of pH in control group UTI group, GSH group and UTI &GSH group shows P<0.001, PCO₂ from lung venous blood gas analysis shows P<0.005, PO₂ shows P<0. 005, PAP shows P<0.001, which all have statistical significance. 2.MDA measurement, MPO content: the comparison of MDA measurement, MPO content in control group UTI group, GSH group and UTI &GSH group has statistical significance (P<0.001) .3. W/D of lung tissue: the comparison of the lung tissue W/D in control group, UTI group, GSH group and UTI&GSH group shows statistical significance (P<0.001) .4. The pathological changes in lung tissue through optical microscope: Lung tissue cell wall is slightly widened, the vascular endothelial is integrity, there are blood-cells in the vessel. But in the control group, pulmonary alveolar wall is widened, accompanied by scattered in the fibrin deposition, vascular endothelial is also integrity, a large number of neutrophils infiltrated, it can be seen a number of alveolar phagocytic cells, accompanied by the neutrophil aggregation.5. The pathological changes in lung tissue through electron microscope: in UTI group, GSH group and UTI&GSH group, most of the alveolar organizational structure is normal, type I and type II alveolar epithelial cells are generally normal, part of alveolar epithelial cells edemaed and fell off. The surface villi is still visible, lamellar body is particularly prevalent, alveolar lumen is basically patent. However, RBC can be seen in a small number of alveolar cavities. In some regional, capillary is filled with erythrocyte, a few inflammatory cells infiltrated can be seen;but in the control group, the capillary in the alveolar septa is hyperaemic obviously, the blood vessel is dilatated, the RBC leakage in alveolar cavity is obvious, deposited in piles, accompanied by the exudation of cellulose, infiltration of neutrophils can be seen, accompanied by a few of monocaryotic cells. Some of the alveolar epithelium cells edemaed and fell off to result alveolar wall became thinner or broken, some of alveolar epithelial cells aqtocytolysis. The lamellar bodyin type I and type II alveolar epithelial cell cytoplasm was emptied.6. The comparison of apoptotic index: compared with UTI group, GSH group and UTI&GSH group, in the control group there are more cells whose nucleus contains a purple blue grain which is a apoptosis cell than those are in the UTI group, GSH group and UTI&GSH group, this shows statistical significance (P< 0.001) .Conclusion:In the models of rabbit lung transplantation, UTI and GHS can effectively reduce the ischemical reperfusion injury of pulmonary grafts, and improve the pulmonary function after the transplantation.