CAJ | 학술논문

目的探讨氧控制性再灌注抗犬体外循环肺缺血再灌注早期损伤过程中高迁移族蛋白1(HMGB1)的表达.方法健康犬14只按完全随机法分为对照组(n=7)和实验组(n=7),均进行体外循环肺缺血再灌注.对照组全程吸入氧浓度(FiO2)80%;实验组在主动脉开放即刻调整FiO2至40%,随后每5 min依次上调10%,最后达80%,其余时段均维持FiO2 80%.观察2组犬在麻醉后、主动脉开放前、主动脉开放后5、10、15、20、25 min以及停机前动脉血氧分压(PaO2)的变化.在开胸后即刻(T1)、主动脉开放25 min(T2)、主动脉开放90min(T3)分别留取血液标本及肺组织标本,检测肺组织HMGB1及NF-κB的表达,酶联免疫吸附法(ELISA)检测血清IL-6和TNF-α含量;检测肺组织湿干质量比及肺组织髓过氧化物酶(MPO)活性、丙二醛(MDA)含量;观察肺组织病理学变化.结果实验组PaO2在主动脉开放后5、10、15、20min均显著低于对照组(均P＜0.05).与对照组比较,在T2和T3时点实验组HMGB1 mRNA表达(T2:0.9260±0.013 9比1.0496±0.0306;T3:0.832 5±0.0154比0.9894±0.0144,均P＜0.05)和蛋白表达(T2:0.434 5±0.074 8比0.551 2±0.047 4;T3:0.449 0±0.054 1比0.545 5±0.040 6,均P＜0.05)均降低.实验组肺组织NF-κB蛋白表达在T2和T3时点低于对照组(均P＜0.05).实验组血清IL-6和TNF-α含量在T2和T3时点低于对照组(均P＜0.05).实验组肺组织湿干质量比、MPO活性、MDA含量在T2和T3时点均低于对照组(均P＜0.05).实验组肺损伤评分在T2和T3时点均低于对照组[T2:(2.0±0.7)分比(3.8±0.5)分;T3:(2.6±0.6)分比(4.2±0.8)分,均P＜0.05].结论氧控制性再灌注对体外循环肺缺血再灌注早期损伤具有保护作用,其机制可能与下调HMGB1表达有关.
목적 탐토양공제성재관주항견체외순배폐결혈재관주조기손상과정중고천이족단백1(HMGB1)적표체.방법 건강견14지안완전수궤법분위대조조(n=7)화실험조(n=7),균진행체외순배폐결혈재관주.대조조전정흡입양농도(FiO2)80%;실험조재주동맥개방즉각조정FiO2지40%,수후매5 min의차상조10%,최후체80%,기여시단균유지FiO2 80%.관찰2조견재마취후、주동맥개방전、주동맥개방후5、10、15、20、25 min이급정궤전동맥혈양분압(PaO2)적변화.재개흉후즉각(T1)、주동맥개방25 min(T2)、주동맥개방90min(T3)분별류취혈액표본급폐조직표본,검측폐조직HMGB1급NF-κB적표체,매련면역흡부법(ELISA)검측혈청IL-6화TNF-α함량;검측폐조직습간질량비급폐조직수과양화물매(MPO)활성、병이철(MDA)함량;관찰폐조직병이학변화.결과 실험조PaO2재주동맥개방후5、10、15、20min균현저저우대조조(균P＜0.05).여대조조비교,재T2화T3시점실험조HMGB1 mRNA표체(T2:0.9260±0.013 9비1.0496±0.0306;T3:0.832 5±0.0154비0.9894±0.0144,균P＜0.05)화단백표체(T2:0.434 5±0.074 8비0.551 2±0.047 4;T3:0.449 0±0.054 1비0.545 5±0.040 6,균P＜0.05)균강저.실험조폐조직NF-κB단백표체재T2화T3시점저우대조조(균P＜0.05).실험조혈청IL-6화TNF-α함량재T2화T3시점저우대조조(균P＜0.05).실험조폐조직습간질량비、MPO활성、MDA함량재T2화T3시점균저우대조조(균P＜0.05).실험조폐손상평분재T2화T3시점균저우대조조[T2:(2.0±0.7)분비(3.8±0.5)분;T3:(2.6±0.6)분비(4.2±0.8)분,균P＜0.05].결론 양공제성재관주대체외순배폐결혈재관주조기손상구유보호작용,기궤제가능여하조HMGB1표체유관.
Objective To observe the expression of high mobility group box 1 (HMGB1)in the lung protection of oxygen controlled reperfusion against ischemia-reperfusion (I/R) with cardiopulmonary bypass (CPB) in canine. Methods Fourteen healthy canines were randomly divided into two groups:control group (n=7) and test group (n=7), and received CPB. The animals in the control group received 80% FiO2 throughout the whole procedure, whereas those in the test group received 40% immediately at the declamping of aorta, and an additional 10% every 5 min later until 80% FiO2 was reached. Other procedures were the same with control group. Arterial oxygen partial pressure (PaO2) was recorded after anesthesia was completed, before and at 5, 10, 15, 20, 25 min after sternotomy, and before withdrawal of CPB. HMGB1 expression by RT-PCR and Western blot, NF-κB expression by Western blot, IL-6 and TNF-α by ELISA were analyzed just after sternotomy(T1 ), 25 min after declamping(T2) and 90 min after declamping(T3). At the same time points, malondialdehyde, myeloperoxidase activity, wet/dry mass (Mw/MD) gnd tissue pathology were analyzed. Results PaO2 was significantly lower in the test group than that in the control group at 5, 10, 15 and 20 min after aortic declamping (all P＜0.05). Compared with the control group at T2 and T3, the test group was found to have lower levels in HMGB1 mRNA and protein expressions (T2:0.926 0±0.013 9 vs 1.049 6±0.030 6;T3:0.832 5±0.015 4 vs 0.989 4±0.014 4, both P＜0.05; T2:0.434 5±0.074 8 vs 0.551 2±0.047 4;T3:0.449 0±0.054 1vs 0.545 5±0.040 6, both P＜0.05), NF-κB protein expression in the lung tissues, IL-6 and TNF-α in serum, Mw/MD ratio, MDA level and MPO activity (all P＜0.05).Pathological scores in test group at T2 and T3 were significantly lower than those in control group (T2:2.0±0.7vs 3.8 ±0.5; T3:2.6±0.6 vs 4.2 ±0.8, both P＜0.05). Conclusion Oxygen controlled reperfusion possesses lung protection in early stage of I/R injury with CPB, which may be related to its inhibition of HMGB1 expression.