【摘要】 目的:建立大鼠的原位异体左单肺移植模型。比较乌司他丁(ulinastatin,UTI)和非抗凝肝素氮脱硫酸肝素(non-anticoagulant N-desulfated heparin,NNH)在该模型中对移植肺缺血再灌注损伤的保护作用。方法:纯种雄性SD大鼠24只,随机分成分为三组, 再灌注时给予生理盐水对照组(n=8):1．2 ml／kg;UTI治疗组(n=8):10,000 U／kg;NNH治疗组(n=8):12 mg／kg,在显微技术和机械通气下完成左单肺移植。再灌注2 h后测量移植肺肺静脉血气,肺干湿重比W／D,髓过氧化物酶(MPO)浓度,TNF-α和IL-8浓度, 细胞间粘附分子-1(ICAM-1)的mRNA表达,肺组织病理切片。结果:24只移植鼠至实验结束存活21只,存活率为87．5％,套管成功率为100％。3只失败大鼠中2只为移植再灌注后分别于75与90 min左右死亡,1只大鼠仍存活,但开胸取肺时见肺动脉和静脉内血栓形成,移植肺无灌注。供肺肺静脉血气PaO2 UTI组(289．68±5．65 mmHg)及NNH组(315．26±14．91 mmHg)与对照组(236．80 ±13．90 mmHg)均有显著差异(P<0．0001),NNH组和UTI组PaO2也有显著差异(P<0．0001);UTI组干湿重比W／D(4．74± 0．28)、NNH组(4．87±0．34)和对照组(5．00±0．12)无显著差异(P=0．18);UTI组MPO活性(0．52±0．25)低于对照组(0．84±0．22,P <0．05),NNH组MPO活性(0．64±0．24)和对照组(P=0．12)、UTI组(P=0．29)均无显著差异;肺组织中TNF-α浓度UTI组(138．61 ±15．30)显著高于对照组(77．54±10．14,P<0．0001),NNH组(52．85±6．55)显著低于对照组(P<0．0001);肺组织IL-8浓度UTI组 (173．45±27．63)显著高于对照组(76．22±12．42,P<0．0001)和NNH组(61．34±9．86,P<0．0001),NNH组和对照组无显著差异(P =0．12)。ICAM-1 mRNA表达UTI组(0．37±0．14)、NNH组(0．38±0．11)显著低于对照组(1．74±0．39,P<0．0001),UTI组和NNH 组无显著差异(P=0．94);光镜下病理示UTI和NNH组肺组织损伤轻于对照组。结论:非内皮化袖套大鼠左单肺移植动物模型用于观察时间较短的再灌注损伤的研究是合乎要求的。UTI和NNH对大鼠肺移植后缺血再灌注损伤具有相似的保护作用,UTI具有较强的清除氧自由基作用,NNH减少了致炎细胞因子的表达,改善移植肺的功能,对减轻肺缺血再灌注损伤较UTI有更明显的保护作用。更多还原

【Abstract】 Objective: To build an orthotopic single left lung allograft animal model in rats and compare the effects of ulinastatin (UTI) and non-anticoagulant N-desulfated heparin (NNH) on lung ischemia-reperfusion injury after lung transplantation. Methods: 24 Male SD rats weighted 250-350g were randomly divided into 3 groups. At the time of reperfusion, the control group were given saline 1. 2ml/kg (n = 8); the UTI group were given UTI 10,000 U/kg (n=8); the NNH group were given NNH 12mg/kg , we completed 24 rat left lung transplantation assisted by mechanical ventilation and microscopy. 2 hours after reperfusion, the transplanted lung’s pulmonary vein blood gases, wet dry ratio(W/D) and myeloperoxidase(MPO) were measured. Tumor necrosis factor-α(TNF-α) and interleukin 8(IL-8) were measured by method of ELISA. Intercilluar adhesion molecule-1( IC AM-1) mRNA in lung tissue were measured by method of RT-PCR, pathology slice were done routinely. Results: Among 24 transplanted rats, 21 survived at the end point, the transplantation success rate was 87. 5% and the cuff procedure success rate was 100%. Among the 3 failure cases, 2 died in 75 minutes and 90 minutes after reperfusion respectively. Another one was still alive, however, thrombus was found in the pulmonary artery and vein at autopsy, thus the transplanted lung was not perfused. PaO2 in the UTI group(289. 68±5. 65mmHg) and the NNH group(315. 26 ± 14. 91mmHg) was significantly higher (P<0.0001) than the control group (236. 80 ±13. 90mmHg ), and the NNH group was singnificantly higher than the UTI group(P< 0.0001). W/D ratio has no difference(P=0.18) within the UTI group(4.74 ±0. 28) ,the NNH group (4. 87 ±0. 34) and the control group (5. 00 ± 0. 12), MPO activity in the UTI group(0. 52 ± 0. 25) was lower than the control group(0. 84±0. 22, P<0.05), and the difference within the NNH group(0. 64 ± 0. 24), the control group and the UTI group was not apparent(P=0. 12, P=0. 29). In the lung tissue, the concentrations of TNF-a in the UTI group (138. 61 ± 15. 30) were significantly higher than the control group(77. 54 ± 10.14, P<0. 0001), the NNH group(52. 85 ± 6. 55) was significantly lower than the control group(P<0.0001). In the lung tissue, the concentrations of IL-8 in UTI group(173. 45± 27. 63) were significantly higher than the control group(76. 22 ?12. 42, P<0.0001) and the NNH group(61. 34 ± 9. 86,P<0. 0001), and there was no difference between the control group and the NNH group(P=0. 12). ICAM-1 mRNA in the UTI group(0. 37 ± 0. 14) and the NNH group(0. 38 ± 0. 11 )5. 00 ± 0. 12, P<0. 01 and P<0. 0001) were significantly lower than the control group(1. 74 ±0. 39,P<0. 0001), and there was no difference between the UTI group and the NNH group(P=0. 94). Histological appearances showed less lung tissue damage in the UTI and NNH group. Conclusion: Rat left lung transplantation model with "non-vessel endo-thelium cuff" technique is easy to do and acceptable for short period research. UTI and NNH both prevented lung reperfusion injury after lung transplantation in rats. UTI can significantly reduce oxygen derived free radicals. NNH depressed the expression of proinflammatory fator and improved the function of transplanted lung, and have more significantly effect on preventing the lung reperfusion injury after lung transplantation in rats.更多还原