中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2015年
8期
1798-1801
,共4页
王克胜%王晓雷%金延武%张贺%赵鑫
王剋勝%王曉雷%金延武%張賀%趙鑫
왕극성%왕효뢰%금연무%장하%조흠
肺缺血%再灌注损伤%缺氧诱导因子-1α%诱导型一氧化氮合酶
肺缺血%再灌註損傷%缺氧誘導因子-1α%誘導型一氧化氮閤酶
폐결혈%재관주손상%결양유도인자-1α%유도형일양화담합매
Lung ischemia%Reperfusion injury%Hypoxia inducible transcription factor-1α%Inducible nitric oxide synthase
目的 探讨缺氧诱导因子(HIF)-1α-诱导型一氧化氮合酶(iNOS)-一氧化氮(NO)信号通路在肺缺血再灌注损伤(LIRI)中的作用及其机制.方法 选择200 ～ 250 g的健康雄性SD大鼠40只,随机分为5组(n=8):假手术组(C组)、缺血再灌注组(I/R组)、缺血再灌注加二甲基乙二酰基甘氨酸(DMOG)处理组(DMOG组)、缺血再灌注加诱导型一氧化氮合酶(iNOS)抑制剂1 400W处理组(1 400W组)、缺血再灌注加DMOG和1400W处理组(DW组).C组:仅行左侧开胸,不行缺血再灌注处理;I/R组:建立大鼠LIRI模型,夹闭左侧肺门,阻断60 min后,再灌注120 min;DMOG组和DW组:在动物模型建立前24 h腹腔注射DMOG 40 mg/kg;1 400 W组和DW组在实验开始麻醉后即刻腹腔注射1 400 W 5 mg/kg.实验结束后,留取各组大鼠左肺组织标本.严格按照试剂盒操作步骤检测肺组织HIF-1 α蛋白表达,iNOS、eNOS和SOD活性,NO、丙二醛(MDA)和白细胞介素(IL)-8含量;计算肺组织湿/干重之比(W/D).结果 (1)与C组比较,I/R、DMOG、1 400W和DW组HIF-1α蛋白表达(154.61 ±6.56、138.23±10.81、151.68 ±9.65、136.35±5.78比178.72 ±8.34)增加,eNOS活性[(0.35±0.09)、(0.32 ±0.10)、(0.50 ±0.13)、(0.49±0.15)比(0.58 ±0.12) U/mg·蛋白]降低;I/R组和DMOG组iNOS活性[(0.91±0.27)、(1.15 ±0.32)比(0.41±0.13) U/mg·蛋白]和NO含量[(0.87 ±0.18)、(1.03 ±0.16)比(0.44 ±0.09)μmol/L]增加(P＜0.05).(2)与I/R组比较,DMOG组和DW组HIF-1α蛋白表达(138.23±10.81、136.35±5.78比154.61 ±6.56)增加;1400W组和DW组iNOS活性[(0.39 ±0.11)、(0.42 ±0.12)比(0.91±0.27) U/mg·蛋白]和NO含量[(0.41 ±0.11)、(0.40 ±0.08)比(0.87 ±0.18) μmol/L]降低,eNOS活性[(0.50±0.13)、(0.49 ±0.15)比(0.35 ±0.09) U/mg·蛋白]增加(P＜0.05).结论 HIF-1α-iNOS-NO信号通路参与了LIRI的过程,其可能机制是:缺血再灌注上调HIF-1α蛋白的表达,通过iNOS/NO通路引起肺组织损伤.
目的 探討缺氧誘導因子(HIF)-1α-誘導型一氧化氮閤酶(iNOS)-一氧化氮(NO)信號通路在肺缺血再灌註損傷(LIRI)中的作用及其機製.方法 選擇200 ～ 250 g的健康雄性SD大鼠40隻,隨機分為5組(n=8):假手術組(C組)、缺血再灌註組(I/R組)、缺血再灌註加二甲基乙二酰基甘氨痠(DMOG)處理組(DMOG組)、缺血再灌註加誘導型一氧化氮閤酶(iNOS)抑製劑1 400W處理組(1 400W組)、缺血再灌註加DMOG和1400W處理組(DW組).C組:僅行左側開胸,不行缺血再灌註處理;I/R組:建立大鼠LIRI模型,夾閉左側肺門,阻斷60 min後,再灌註120 min;DMOG組和DW組:在動物模型建立前24 h腹腔註射DMOG 40 mg/kg;1 400 W組和DW組在實驗開始痳醉後即刻腹腔註射1 400 W 5 mg/kg.實驗結束後,留取各組大鼠左肺組織標本.嚴格按照試劑盒操作步驟檢測肺組織HIF-1 α蛋白錶達,iNOS、eNOS和SOD活性,NO、丙二醛(MDA)和白細胞介素(IL)-8含量;計算肺組織濕/榦重之比(W/D).結果 (1)與C組比較,I/R、DMOG、1 400W和DW組HIF-1α蛋白錶達(154.61 ±6.56、138.23±10.81、151.68 ±9.65、136.35±5.78比178.72 ±8.34)增加,eNOS活性[(0.35±0.09)、(0.32 ±0.10)、(0.50 ±0.13)、(0.49±0.15)比(0.58 ±0.12) U/mg·蛋白]降低;I/R組和DMOG組iNOS活性[(0.91±0.27)、(1.15 ±0.32)比(0.41±0.13) U/mg·蛋白]和NO含量[(0.87 ±0.18)、(1.03 ±0.16)比(0.44 ±0.09)μmol/L]增加(P＜0.05).(2)與I/R組比較,DMOG組和DW組HIF-1α蛋白錶達(138.23±10.81、136.35±5.78比154.61 ±6.56)增加;1400W組和DW組iNOS活性[(0.39 ±0.11)、(0.42 ±0.12)比(0.91±0.27) U/mg·蛋白]和NO含量[(0.41 ±0.11)、(0.40 ±0.08)比(0.87 ±0.18) μmol/L]降低,eNOS活性[(0.50±0.13)、(0.49 ±0.15)比(0.35 ±0.09) U/mg·蛋白]增加(P＜0.05).結論 HIF-1α-iNOS-NO信號通路參與瞭LIRI的過程,其可能機製是:缺血再灌註上調HIF-1α蛋白的錶達,通過iNOS/NO通路引起肺組織損傷.
목적 탐토결양유도인자(HIF)-1α-유도형일양화담합매(iNOS)-일양화담(NO)신호통로재폐결혈재관주손상(LIRI)중적작용급기궤제.방법 선택200 ～ 250 g적건강웅성SD대서40지,수궤분위5조(n=8):가수술조(C조)、결혈재관주조(I/R조)、결혈재관주가이갑기을이선기감안산(DMOG)처리조(DMOG조)、결혈재관주가유도형일양화담합매(iNOS)억제제1 400W처리조(1 400W조)、결혈재관주가DMOG화1400W처리조(DW조).C조:부행좌측개흉,불행결혈재관주처리;I/R조:건립대서LIRI모형,협폐좌측폐문,조단60 min후,재관주120 min;DMOG조화DW조:재동물모형건립전24 h복강주사DMOG 40 mg/kg;1 400 W조화DW조재실험개시마취후즉각복강주사1 400 W 5 mg/kg.실험결속후,류취각조대서좌폐조직표본.엄격안조시제합조작보취검측폐조직HIF-1 α단백표체,iNOS、eNOS화SOD활성,NO、병이철(MDA)화백세포개소(IL)-8함량;계산폐조직습/간중지비(W/D).결과 (1)여C조비교,I/R、DMOG、1 400W화DW조HIF-1α단백표체(154.61 ±6.56、138.23±10.81、151.68 ±9.65、136.35±5.78비178.72 ±8.34)증가,eNOS활성[(0.35±0.09)、(0.32 ±0.10)、(0.50 ±0.13)、(0.49±0.15)비(0.58 ±0.12) U/mg·단백]강저;I/R조화DMOG조iNOS활성[(0.91±0.27)、(1.15 ±0.32)비(0.41±0.13) U/mg·단백]화NO함량[(0.87 ±0.18)、(1.03 ±0.16)비(0.44 ±0.09)μmol/L]증가(P＜0.05).(2)여I/R조비교,DMOG조화DW조HIF-1α단백표체(138.23±10.81、136.35±5.78비154.61 ±6.56)증가;1400W조화DW조iNOS활성[(0.39 ±0.11)、(0.42 ±0.12)비(0.91±0.27) U/mg·단백]화NO함량[(0.41 ±0.11)、(0.40 ±0.08)비(0.87 ±0.18) μmol/L]강저,eNOS활성[(0.50±0.13)、(0.49 ±0.15)비(0.35 ±0.09) U/mg·단백]증가(P＜0.05).결론 HIF-1α-iNOS-NO신호통로삼여료LIRI적과정,기가능궤제시:결혈재관주상조HIF-1α단백적표체,통과iNOS/NO통로인기폐조직손상.
Objective To investigate the role of hypoxia inducible factor (HIF)-1 α-inducible nitric oxide synthase (iNOS)-nitric oxide (NO) signal pathway in lung ischemia-reperfusion (I/R) injury and the underlying mechanisms.Methods A total of 40 male Sprague-Dawley rats (200-250 g) were randomly divided into control (C group),I/R group,I/R plus HIF-1α stablilizer DMOG (DMOG group),I/R plus iNOS inhibitor 1 400 W (1 400 W group) and I/R plus DMOG and 1 400 W group (DW group) (n =8 each).Animals in DMOG group and DW group were given an intraperitoneal injection of DMOG (40 mg/kg in 1 ml saline).Animals in 1 400 W group and DW group were given an intraperitoneal injection of 1 400 W(5 mg/kg in 1 ml saline) after anesthesia.At the end of the experiment,the left lung was harvested.The protein expression of HIF-1 α,the activity of iNOS,eNOS and superoxide dismutase (SOD),and the levels of NO,malondialdehyde (MDA) and interleukin (IL)-8 were detected.Results (1) As compared with C group,the protein expression levels of HIF-1 α in L/R group,DMOG group,1 400 W group,and DW group (154.61 ±6.56,138.23 ± 10.81,151.68 ±9.65,136.35 ±5.78 vs.178.72 ± 8.34) were increased,the activity of eNOS [(0.35 ± 0.09),(0.32 ± 0.10),(0.50 ± 0.13),(0.49 ±0.15) vs.(0.58 ±0.12) U/mg·prot] was decreased;the activity of iNOS in I/R group and DMOG group [(0.91 ±0.27),(1.15 ±0.32) vs.(0.41 ±0.13) U/mg·prot] and the level of NO [(0.87±0.18),(1.03±0.16) vs.(0.44 ±0.09) μmol/L] were increased (P＜0.05).(2) As Compared with I/R group,the protein expression levels of HIF-1α in DMOG group and DW group (138.23 ± 10.81,136.35 ± 5.78 vs.154.61 ± 6.56) were increased;the activity of iNOS in 1 400 W group and DW group [(0.39 ±0.11),(0.42 ±0.12) vs.(0.91 ±0.27) U/mg·prot] and the level of NO [(0.41 ±0.11),(0.40 ±0.08) vs.(0.87 ±0.18) μ mol/L] were decreased,and the activity of eNOS [(0.50 ±0.13),(0.49 ±0.15) vs.(0.35 ±0.09) U/mg·prot] was increased (P＜0.05).Conclusion HIF-1α-iNOS-NO signal pathway contributes to lung I/R injury,and the mechanisms might be that I/R up-regulates the protein expression of HIF-1α,and induces lung injury through iNOS/NO pathway.
