Kang Ming-qiang, Lin Pei-qiu, Lin Ruo-bai, Huang Xue-shan, Lin Xu, Zheng Yu-hui, Liao Chong-xian, Chen Dao-zhong
Department of Thoracic Surgery, Union Hospital Affiliated to Fujian Medical University, Fuzhou 350001, China.
Zhonghua Yi Xue Za Zhi. 2007 Dec 25;87(48):3425-8.
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).
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.
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.
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.
探讨经肺静脉将腺病毒介导的人白细胞介素10（hIL-10）基因转导入肺同种异体移植供肺的可行性，以及hIL-10基因转导对后续缺血再灌注损伤（IRI）的影响。
56只雄性SD大鼠随机分为4组，每组14只：D组，采用改良套袖吻合技术行左肺同种异体移植（经血管转染5 ml含5×10⁹ 空斑形成单位/ml腺病毒载体-hIL-10复合物）；C组，经血管转染空白腺病毒载体Adenovec；B组，经血管转染稀释剂；A组，行假手术。所有供肺在移植前于10℃保存3小时。再灌注4小时后，从腹主动脉采集血样进行血气分析，以氧分压（PaO₂）评估肺功能。然后处死大鼠，取出左肺进行病理检查，测量移植肺的湿/干（W/D）重量比。采用SABC免疫组化法检测细胞质中hIL-10的表达，ELISA法检测肿瘤坏死因子-α（TNF-α）和干扰素-γ（IFN-γ）的表达，检测丙二醛（MDA）、超氧化物歧化酶（SOD）和髓过氧化物酶（MPO）的水平，并分析病理形态学变化。
D组的PaO₂水平显著高于B组和C组（均P < 0.01）。D组的W/D比值、MDA和MPO水平显著低于B组和C组（均P < 0.01），但D组的SOD水平显著高于B组和C组（均P < 0.05）。D组的TNF-α和IFN-γ水平显著低于B组和C组（均P < 0.01），肺组织水肿和间质炎症较少。D组大鼠肺组织的RT-PCR显示有hIL-10表达，其他组未检测到。
经肺静脉将腺病毒介导的hIL-10基因转导入肺同种异体移植供肺是可行且有效的。hIL-10基因转导至肺同种异体移植供肺可减轻后续IRI，改善移植后早期移植肺功能。
