| With the developing of the medical technology,organ transplantation is the most effective therapeutic treatment for chronic end-stage diseases.The number of lung transplantation patients is growing every year,but compared with the transplantion of other organs,long-term survival rate is still not ideal.Statistics show that 5-year survival rate is only 45% of lung transplantation.70% of patients progress to occlusive bronchiolitis after 5 years with lung transplantion.Clinical and experimental studies have shown that occlusive bronchiolitis which is also called lung transplantation chronic reaction,which restricts the long-term survival quality of lung transplantation patients.Therefore,how to reduce and treat the lung transplantation related occlusive bronchiolitis has become an important subject of present research.HO-1 is one kind of protective genes in the organ transplantation,we use the cell protective activity of HO-1 to significantly improve cell damage which is caused by schemia reperfusion,acute rejection of lung transplantation,and improve the recovery of function after reperfusion organ transplantation.This subject is divided into the following two parts:Part one: Establish an effective lung transplantion related occlusive bronchiolitis model.Objective: Study and establish a more stable and effective rat lung transplant related occlusive bronchiolitis model.Methods: 30 rats were randomly divided into 2 groups,one was homologous transplantation,the other was allogeneic transplantation.Preliminary analysis was done for the situation of immune rejection,such as the damage of epithelial,tracheal glands,cartilage ring and interstitial tissue.Results: The group of allogeneic transplantation appeared edema of trachea epithelium,gland,cartilage ring at the beginning of experiment.And then interstitial tissue,hematoma and infiltration of lymphocytes,monocytes.necrosis of granulation tissue reaction is found after 60 days.The tracheal structure of homologous transplantation group was relatively intact,smooth,and there was no percolate in the tracheal cavity and stroma.Conclusion: Allogeneic transplantation group is more suitable for making the model of the rat lung transplantation realted occlusive bronchiolitis.Part two: The protection of HO-1 in rat lung transplantation related occlusive bronchiolitisObjective: Establish a model of rat lung transplantation related occlusive bronchiolitis,research the protection of HO-1 in rat lung transplantation related occlusive bronchiolitis.Methods: The experimental animals were divided into two group,one was control group which was called homologous transplantation group(SD to SD),the other was experimental group(Wistar to SD)which was stroke-physiological saline solution,Cyclosporin A and adeno-associated virus cantain hemeoxygen-1 gene recombinant of rat.After day 7,30,and 60,the pathological changes,the quantity of CD4~+,CD8~+T around peripheral blood and the apptosis indicators were detected.Results: 1.After day 7 of transplantation,there was no significantly statistical differences of CD4~+T lymphocytes in the following groups,namely homologous transplantation group,saline group,Cs A group and AAV-r HO-1 group(F = 0.799,P =0.512);after day 30 of transplantation there was significantly statistical differences of CD4~+T lymphocytes in the following groups,namely homologous transplantation group,saline group,Cs A group and AAV-r HO-1 group(F=16.630,P=0.000).There was no significant statistical difference during Cs A group and AAV-r HO-1 group,as well as saline group and homologous transplantation group(10.24±1.36 vs.8.76±0.70,P=0.253;16.06±1.36 vs.15.22±3.39,P=0.512).There was significantly statistical differences of CD4~+T lymphocytes in AAV-r HO-1 group compare with homologous transplantation group and normal saline group(16.06±1.36 vs.8.76±0.70,P=0.000;15.22±3.39 vs.8.76±0.70,P=0.000).After day 60 of transplantation,there was significantly statistical differences of CD4~+ T lymphocytes in the following groups,namely homologous transplantation group,saline group,Cs A group and AAV-r HO-1group(F=4.818,P=0.014).There was no significantly statistical differences between homologous transplantation group and normal saline group(P = 0.354),but AAV-r HO-1 group was significant decline,compared with homologous transplantation group,saline group and the Cs A group.The difference was statistically significant(8.09±1.70 vs.12.54±4.13,P=0.015;8.09±1.70 vs.10.98±1.52,P=0.026;8.09±1.70 vs.14.02±2.1;P=0.002)。2.After day 7 of transplantation,there was significantly statistical differences of CD8~+T lymphocytes in the following groups,namely homologous transplantation group,saline group,Cs A group and AAV-r HO-1 group(F=3.804,P=0.031).There was no significant statistical difference between Cs A group and AAV-r HO-1 group,the same with normal saline group and homologous transplantation group(13.02±1.44 vs.13.82±2.90,P=0.557;11.30±0.72 vs.9.69±2.61,P=0.216),but there was significantly statistical differences of CD8~+ T lymphocytes between AAV-r HO-1group and homologous transplantation group,the same with normal saline group(13.02±1.44 vs.9.69±2.61,P=0.024;13.82±2.90 vs.9.69±2.61,P=0.007).After day30 of transplantation,there was significantly statistical differences of CD8~+T lymphocytes in the following groups,namely homologous transplantation group,saline group,Cs A group and AAV-r HO-1 group(F=2.005,P=0.154).There was no significant difference during AAV-r HO-1 group and saline group,the same with the Cs A group(F=0.680;P=0.088),but the AAV-r HO-1 group was significant rise when compare with homologous transplantation group(15.40±1.65 vs.13.12±1.59,P=0.037).After day 60 of transplantation,there was significantly statistical differences of CD8~+T lymphocytes in the following groups,namely homologous transplantation group,saline group,Cs A group and AAV-r HO-1 group(F=2.612,P=0.087).The AAV-r HO-1group had no significantly statistical differences of CD8~+ T lymphocytes when compare with homologous transplantation group and normal saline group(P=0.680;P=0.088),but CD8~+ T was obviously reduced compared to Cs A group(13.84±1.70 vs.17.76±2.95,P=0.028).3.After day 7 of transplantation,there was significantly statistical differences of apoptosis index in the following groups,namely homologous transplantation group,saline group,Cs A group and AAV-r HO-1 group(F=34.219,P=0.000).There was no significantly statistical differences between AAV-r HO-1 group and homologous transplantation group(0.110±0.027 vs.0.086±.0523,P=1.000),but the apoptosis index in AAV-r HO-1 group obviously declined compared to the saline group and Cs A group(0.300±0.027 vs.0.110±0.027,P=0.000;0.244±0.045 vs.0.110±0.027,P=0.000).After day 30 of transplantation,there was significantly statistical differences of apoptosis index in the following groups,namely homologous transplantation group,saline group,Cs A group and AAV-r HO-1 group(F=28.292,P=0.000).There was no significantly statistical differences between AAV-r HO-1 group and homologous transplantation group(0.162±0.041 vs.0.138±0.034,P=1.000),but the apoptosis index in AAV-r HO-1 group obviously declined compared to the saline group and Cs A group(0.424±0.069 vs.0.162±0.041,P=0.000;0.300±0.070 vs.0.162±0.041,P=0.008);After day 60 transplantation,there was significantly statistical differences of apoptosis index in the following groups,namely homologous transplantation group,saline group,Cs A group and AAV-r HO-1group(F=28.947,P=0.000).There was no significantly statistical differences between AAV-r HO-1 group and homologous transplantation group(0.214±0.011 vs.0.276±0.052,P=0.505),but the apoptosis index in AAV-r HO-1 group obviously declined compared to the saline group and Cs A group(0.510±0.059 vs.0.214±0.011,P=0.000;0.358±0.071 vs.0.214±0.011,P=0.003).Conclusion: 1.The inhibitory effect of HO-1 on CD4~+T lymphocyte in the acute phase of cellular immune rejection is not obvious,but the inhibitory effect is significant on CD4 + T lymphocyte in the chronic phase of cellular immune rejection,which is similar to Cs A used by clinic.2.The inhibitory effect of HO-1 on CD8~+T lymphocyte in both acute and chronic phase of cellular immune rejection are obvious,but the inhibitory activity is more stronger than Cs A used by clinic in chronic phase of cellular immune rejection.3.HO-1 can inhibit cells death,and reduce the death of epithelial cells of lung transplantion. |
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