NF-κB抑制剂对大鼠移植肺缺血再灌注损伤的保护作用

Protection of nuclear factor kappaB inhibitor PDTC from ischemia reperfusion injury of rats transplanted lung

目的通过建立大鼠肺移植缺血再灌注模型,检测缺血再灌注损伤中肺组织中NF-κB、TNF-α及ICAM-1的表达,探讨核因子-κB抑制剂PDTC对大鼠肺移植缺血再灌注损伤的影响及其作用机制。方法实验分3组,即假手术对照组(A组)、空白对照组(B组)和PDTC组(C组),每组8例。空白对照组用4℃LPDG液灌洗并保存4 h,PDTC组用含PDTC(40 mmol/L)的4℃LPDG液灌洗并保存供肺4 h。移植肺再灌注1 h后结束实验。测定肺组织湿-干重比、MPO活性,观察移植肺组织病理改变,ELISA测定TNF-α表达水平,免疫组化染色检测ICAM-1及NF-κB表达,RT-PCR检测NF-κB mRNA的水平。结果与空白对照组比较,PDTC组肺组织损伤程度明显减轻,肺组织湿干重比、MPO活性均有下降(P<0.01)。NF-κB、ICAM-1及TNF-α的表达水平较空白对照组明显减少(P<0.01)。结论核因子-κB抑制剂PDTC能有效地减轻缺血再灌注损伤,明显改善移植肺功能,其保护作用可能与抑制TNF-α和ICAM-1的表达有关。

【Objective】To detect the expression of NF-κB, ICAM-1 and TNF-α in rats transplanted lung through rat orthotopic left pulmonary allograft transplantation models. To discuss the protection and mechanisms of nuclear factor kappaB inhibitor PDTC in transplanted rats lung from ischemia reperfusion injury.【Methods】This study was divided into three groups： sham operation control group（A group）, empty control group（B group） and PDTC group（C group）. Every group had 8 cases. The harvested lung blocks were flushed and stored in 4℃ low-potassium-dextran and glucose（LPDG） solution for 4 hours in B group. 4℃ LPDG solution which contains PDTC（40 mmol/L） was used in C group. All animals were killed after lungs were reperfused for 1 hour. The transplanted lung tissue wet-to-dry（W/D） ratio and activity of myeloperoxidase（MPO） and were measured. The pathological changes were observed by the H-E staining. TNF-α was detected by ELISA. The protein expression of NF-κB and ICAM-1 were detected by immunohistochemical staining. NF-κB mRNA was detected by RT-PCR.【Results】After reperfusion the W/D, MPO and the expression of NF-κB, ICAM-1 and TNF-α were decreased evidently in PDTC group than in empty control group. Compared with empty control group, ischemia-reperfusion lung injury was lessened obviously in PDTC group.【Conclusions】Nuclear factor kappaB inhibitor PDTC can effectively ameliorate IRI and further improve transplanted lung functions. The protection mechanism probably is associated with the inhibition of TNF-α and ICAM-l expression.