中国危重病急救医学
中國危重病急救醫學
중국위중병급구의학
CHINESE CRITICAL CARE MEDICINE
2012年
2期
111-115
,共5页
缺血/再灌注损伤,肺%N-乙酰半胱氨酸%天冬氨酸特异性半胱氨酸蛋白酶3%凋亡
缺血/再灌註損傷,肺%N-乙酰半胱氨痠%天鼕氨痠特異性半胱氨痠蛋白酶3%凋亡
결혈/재관주손상,폐%N-을선반광안산%천동안산특이성반광안산단백매3%조망
Lung ischemia/reperfusion injury%N-acetvlcvsteine%Caspase-3%Apoptosis
目的 研究N-乙酰半胱氨酸(NAC)对大鼠肺缺血/再灌注损伤(LIRI)中细胞凋亡及天冬氨酸特异性半胱氨酸蛋白酶3(caspase-3)表达的影响,并探讨其可能机制.方法 雄性SD大鼠42只,按随机数字表法均分为假手术组、LIRI组(阻断肺门45 min后再开放)、NAC组(于LIRI前腹腔注射NAC 150 mg/kg)3组,各组分别于3h、6h取肺组织,用膜联蛋白V-异硫氰酸荧光素(Annexin V-FITC)和碘化丙啶(PI)双染法通过流式细胞仪检测凋亡细胞并计算凋亡率,测定丙二醛(MDA)浓度(硫代巴比妥法)和超氧化物歧化酶(SOD)活性(黄嘌呤氧化酶法),用逆转录-聚合酶链反应(RT-PCR)技术检测caspase-3 mRNA表达,电镜下观察肺脏超微结构的改变.结果 与假手术组相比,LIRI组3h、6h肺组织细胞凋亡率明显增加[(25.60±3.22)％比(2.19±0.48)％,(26.01±4.50)％比(2.55±0.36)％],MDA浓度(nmol/mg)升高(3.26±0.32比0.73±0.23,3.53±0.46比1.08±0.42),SOD活性(U/mg)下降(32.80±3.82比60.51±6.81,33.44±3.24比64.19±6.60),肺组织caspase-3 mRNA表达显著增加(0.717±0,037比0.216±0.046,0.744±0.046比0.227±0.037),差异均有统计学意义(均P＜0.01);与LIRI组相比,NAC组3h、6h肺组织细胞凋亡率明显减少[(14.42±1.61)％比(25.60±3.22)％,( 10.02±1.64)％比(26.01±4.50)％],MDA浓度(nmol/mg)明显下降(1.75±0.33比3.26±0.32,2.15±0.25比3.53±0,46),SOD活性(U/mg)明显升高(42.76±2.06比32.80±3.82,44.94±3.11比33.44±3.24),肺组织caspase-3 mRNA表达显著减少(0.441±0.038比0.717±0.037,0.410±0.037比0.744±0.046),差异均有统计学意义(均P＜0.01).NAC组肺组织超微结构损害较LIRI组明显减轻.在LIRI 3 h、6h时,caspase-3与肺组织细胞凋亡率、MDA呈显著正相关,与SOD呈显著负相关(3h:r值分别为0.9036、0.9216、-0.9511,6h:r值分别为0.9655、0.9650、-0.9574,均P＜0.01).结论 在LIRI早期,NAC可下调caspase-3表达从而抑制细胞凋亡的发生,起到保护肺组织的作用.
目的 研究N-乙酰半胱氨痠(NAC)對大鼠肺缺血/再灌註損傷(LIRI)中細胞凋亡及天鼕氨痠特異性半胱氨痠蛋白酶3(caspase-3)錶達的影響,併探討其可能機製.方法 雄性SD大鼠42隻,按隨機數字錶法均分為假手術組、LIRI組(阻斷肺門45 min後再開放)、NAC組(于LIRI前腹腔註射NAC 150 mg/kg)3組,各組分彆于3h、6h取肺組織,用膜聯蛋白V-異硫氰痠熒光素(Annexin V-FITC)和碘化丙啶(PI)雙染法通過流式細胞儀檢測凋亡細胞併計算凋亡率,測定丙二醛(MDA)濃度(硫代巴比妥法)和超氧化物歧化酶(SOD)活性(黃嘌呤氧化酶法),用逆轉錄-聚閤酶鏈反應(RT-PCR)技術檢測caspase-3 mRNA錶達,電鏡下觀察肺髒超微結構的改變.結果 與假手術組相比,LIRI組3h、6h肺組織細胞凋亡率明顯增加[(25.60±3.22)％比(2.19±0.48)％,(26.01±4.50)％比(2.55±0.36)％],MDA濃度(nmol/mg)升高(3.26±0.32比0.73±0.23,3.53±0.46比1.08±0.42),SOD活性(U/mg)下降(32.80±3.82比60.51±6.81,33.44±3.24比64.19±6.60),肺組織caspase-3 mRNA錶達顯著增加(0.717±0,037比0.216±0.046,0.744±0.046比0.227±0.037),差異均有統計學意義(均P＜0.01);與LIRI組相比,NAC組3h、6h肺組織細胞凋亡率明顯減少[(14.42±1.61)％比(25.60±3.22)％,( 10.02±1.64)％比(26.01±4.50)％],MDA濃度(nmol/mg)明顯下降(1.75±0.33比3.26±0.32,2.15±0.25比3.53±0,46),SOD活性(U/mg)明顯升高(42.76±2.06比32.80±3.82,44.94±3.11比33.44±3.24),肺組織caspase-3 mRNA錶達顯著減少(0.441±0.038比0.717±0.037,0.410±0.037比0.744±0.046),差異均有統計學意義(均P＜0.01).NAC組肺組織超微結構損害較LIRI組明顯減輕.在LIRI 3 h、6h時,caspase-3與肺組織細胞凋亡率、MDA呈顯著正相關,與SOD呈顯著負相關(3h:r值分彆為0.9036、0.9216、-0.9511,6h:r值分彆為0.9655、0.9650、-0.9574,均P＜0.01).結論 在LIRI早期,NAC可下調caspase-3錶達從而抑製細胞凋亡的髮生,起到保護肺組織的作用.
목적 연구N-을선반광안산(NAC)대대서폐결혈/재관주손상(LIRI)중세포조망급천동안산특이성반광안산단백매3(caspase-3)표체적영향,병탐토기가능궤제.방법 웅성SD대서42지,안수궤수자표법균분위가수술조、LIRI조(조단폐문45 min후재개방)、NAC조(우LIRI전복강주사NAC 150 mg/kg)3조,각조분별우3h、6h취폐조직,용막련단백V-이류청산형광소(Annexin V-FITC)화전화병정(PI)쌍염법통과류식세포의검측조망세포병계산조망솔,측정병이철(MDA)농도(류대파비타법)화초양화물기화매(SOD)활성(황표령양화매법),용역전록-취합매련반응(RT-PCR)기술검측caspase-3 mRNA표체,전경하관찰폐장초미결구적개변.결과 여가수술조상비,LIRI조3h、6h폐조직세포조망솔명현증가[(25.60±3.22)％비(2.19±0.48)％,(26.01±4.50)％비(2.55±0.36)％],MDA농도(nmol/mg)승고(3.26±0.32비0.73±0.23,3.53±0.46비1.08±0.42),SOD활성(U/mg)하강(32.80±3.82비60.51±6.81,33.44±3.24비64.19±6.60),폐조직caspase-3 mRNA표체현저증가(0.717±0,037비0.216±0.046,0.744±0.046비0.227±0.037),차이균유통계학의의(균P＜0.01);여LIRI조상비,NAC조3h、6h폐조직세포조망솔명현감소[(14.42±1.61)％비(25.60±3.22)％,( 10.02±1.64)％비(26.01±4.50)％],MDA농도(nmol/mg)명현하강(1.75±0.33비3.26±0.32,2.15±0.25비3.53±0,46),SOD활성(U/mg)명현승고(42.76±2.06비32.80±3.82,44.94±3.11비33.44±3.24),폐조직caspase-3 mRNA표체현저감소(0.441±0.038비0.717±0.037,0.410±0.037비0.744±0.046),차이균유통계학의의(균P＜0.01).NAC조폐조직초미결구손해교LIRI조명현감경.재LIRI 3 h、6h시,caspase-3여폐조직세포조망솔、MDA정현저정상관,여SOD정현저부상관(3h:r치분별위0.9036、0.9216、-0.9511,6h:r치분별위0.9655、0.9650、-0.9574,균P＜0.01).결론 재LIRI조기,NAC가하조caspase-3표체종이억제세포조망적발생,기도보호폐조직적작용.
Objective To investigate the effect of N-acetvlevsteine (NAC) on apoptosis of pneumocytes and expression of caspase-3 during lung ischemia/reperfusion injury (LIRI) in rats,and to explore the possible role of NAC in pneumocyte apoptosis.Methods Forty-two male Sprague-Dawley rats were randomly divided into three groups:sham operation group,LIRI group (LIRI was produced by 45 minutes of clamping of the pulmonary hilum followed by 3 hours or 6 hours of reperfusion),and NAC group (NAC 150 mg/kg was injected intraperitoneally before LIRI).Lung specimens were harvested 3 hours or 6 hours after LIRI.Apoptosis rate in lung tissue was determined with flow cytometer after AnnexinV-fluorescein isothiocyanate (FITC) and propidium iodide (PI) staining.Malondialdehyde (MDA,thiobarbituric acid) and superoxide dismutase (SOD,xanthineoxidase) of lung tissue were measured.Expression of caspase-3 in lung was determined by reverse transcription-polymerase chain reaction (RT-PCR),and the changes in ultrastructure of lung tissue were observed by electron microscope.Results Compared with that of the sham operation group,apoptosis rate of pulmonary cells was significantly increased at 3 hours and 6 hours in LIRI group [ (25.60 ±3.22)％ vs.(2.19 ± 0.48)％,(26.01 ± 4.50)％ vs.(2.55 ± 0.36)％],the content of MDA (nmol/mg) was significantly increased (3.26 ± 0.32 vs.0.73 ± 0.23,3.53 ± 0.46 vs.1.08 ± 0.42),and the activity of SOD (U/mg) was significantly lowered (32.80 ± 3.82 vs.60.51 ± 6.81,33.44 ± 3.24 vs.64.19 ± 6.60),and the expression of caspase-3 mRNA in lung tissue was significantly up-regulated (0.717 ± 0.037 vs.0.216 ± 0.046,0.744 ± 0.046 vs.0.227 ± 0.037,all P＜0.01 ).Compared with that of the LIRI group,apoptosis rate of pulmonary cell was significantly decreased [ ( 14.42 ±1.61)％ vs. (25.60 ± 3.22)％, (10.02 ± 1.64)％ vs. (26.01 ± 4.50)％],content of MDA (nmol/mg) was lowered significantly ( 1.75 ± 0.33 vs.3.26 ± 0.32,2.15 ± 0.25 vs.3.53 ± 0.46),and activity of SOD (U/mg) was significantly elevated (42.76 ± 2.06 vs.32.80 ± 3.82,44.94 ± 3.11 vs.33.44 ± 3.24,all P＜0.01 ) in NAC group.The expression of caspase-3 in lung tissue was remarkably down-regulated compared with that of LIRI group (0.44 1 ± 0.038 vs.0.717 ±0.037,0.410 ± 0.037 vs.0.744 ± 0.046,both P＜0.01 ).The ultrastructure changes in lung tissue were milder in NAC group than in LIRI group.Positive correlation was found between the expression of caspase-3 and apoptosis rate and the content of MDA (3 hours:r=0.9036,0.9216; 6 hours:r=0.9655,0.9650,all P＜0.01),but negative correlation was found between apoptosis rate and activity of SOD (3 hours:r=-0.9511,6 hours:r=- 0.9574,both P＜0.01 )after LIRI 3 hours and 6 hours.Conclusion During early period of LIRI,caspase-3 was significantly deregulated by NAC,therefore the cellular apoptosis was inhibited,thus protecting lung tissue from LIRI.
