[Isografts on subsequent ischemia-reperfusion injury: experiment with rats].
OBJECTIVE
To investigate the feasibility of ex vivo adenovirus-mediated gene transfer of human interleukin10 (hIL10) via the pulmonary vein into lung isografts, and to investigate the effect of hIL-10 gene transfer on subsequent ischemia-reperfusion injury (IRI).


METHODS
Fifty-six male SD rats were randomly divided into 4 equal groups: Group D, undergoing left lung isotransplantation with the improved cuff anastomosis technique (the Isografts were transvascularly transfected 5 ml of 5 x 10(9) plaque-forming units/ml adenovec-hIL-10 complex, Group C, with the Isografts transvascularly transfected with blank adenovirus vector Adenovec, Group B, with the Isografts transvascularly transfected with diluent , and Group A, undergoing sham operation. All allografts were preserved for 3 hours at 10 degrees C before transplantation. Four hours after reperfusion blood samples were collected from hr abdominal aorta to undergo blood air analysis. Lung function was evaluated by partial pressure of oxygen (PaO2). Then the rats were killed with their left lung taken out to undergo pathological examination. The graft lung wet-to-dry (W/D) weight ratio was measured. SABC immunohistochemistry was used to detect the expression of hIL-10 in the cytoplasm. ELISA was used to detect the expression of tumor necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma). The levels of malonyldialdehyde (MDA), superoxide dismutase (SOD), and myeloperoxidase (MPO) were measured by. Pathological morphologic change was also analyzed.


RESULTS
The PaO2 level of Group D was significantly higher than those of Groups B and C (both P < 0.01). The W/D ratio, and levels of MDA and MPO of Group D were significantly lower than those of Groups B and C (both P < 0.01), but the SOD level of Group D was significantly higher than those of Groups B and C (both P < 0.05). The TNF-alpha and IFN-gamma levels of Group D were significantly lower than those of Groups B and C (both P < 0.01). Fewer tissue edema and interstitial inflammation were found in lungs. Of Group D RT-PCR showed hIL-10 expression in the lungs of the rats of Group D, but not in other groups.


CONCLUSIONS
Ex vivo adenovirus-mediated gene transfer of hIL-10 via the pulmonary vein into the lung isografts is feasible and effective. hIL-10 gene transfer into lung isografts ameliorates subsequent IRI and improves early posttransplant graft function.
Kang
Ming-qiang
MQ
Department of Thoracic Surgery, Union Hospital Affiliated to Fujian Medical University, Fuzhou 350001, China.
Lin
Pei-qiu
PQ
Lin
Ruo-bai
RB
Huang
Xue-shan
XS
Lin
Xu
X
Zheng
Yu-hui
YH
Liao
Chong-xian
CX
Chen
Dao-zhong
DZ
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D004195:Disease Models, Animal; D004797:Enzyme-Linked Immunosorbent Assay; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D007371:Interferon-gamma / Q000378:metabolism; D016753:Interleukin-10 / Q000235:genetics*; D016040:Lung Transplantation / Q000379:methods*; D008297:Male; D008315:Malondialdehyde / Q000378:metabolism; D009195:Peroxidase / Q000378:metabolism; D011897:Random Allocation; D051381:Rats; D017207:Rats, Sprague-Dawley; D015427:Reperfusion Injury / Q000517:prevention & control*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013482:Superoxide Dismutase / Q000378:metabolism; D014162:Transfection / Q000379:methods; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
D014409:Tumor Necrosis Factor-alpha; D016753:Interleukin-10; D008315:Malondialdehyde; D007371:Interferon-gamma; D009195:Peroxidase; D013482:Superoxide Dismutase