CAJ | 학술논문

目的通过建立大鼠机械通气及放射性肺损伤动物模型,研究机械通气对放射性肺损伤大鼠肺组织急性炎症、氧化应激及细胞凋亡的影响.方法 40只雄性Sprague-Dawley(SD)大鼠,采用随机数字表法分成4组,每组10只,即健康对照组、照射组、通气组和照射后通气组.观测各组肺组织病理改变;凝胶电泳迁移率实验(EMSA)检测肺组织细胞NF-κB的活性;Western blot检测肺细胞核内NF-κB亚基P65蛋白表达水平;采用DNA断端末端标记(TUNEL)法检测肺细胞的凋亡;同时检测肺湿干重比值(W/D)、髓过氧化物酶(MPO)、丙二醛(MDA)、超氧化物歧化酶(SOD);检测肺灌洗液中总蛋白、白细胞计数的水平.结果与健康对照组相比,其余3组急性肺损伤(ALI)评分、W/D比值、MPO活性、总蛋白、白细胞计数、凋亡指数(AI)、肺组织MDA值、NF-κB的活性及P65蛋白的表达均显著性增高,差异有统计学意义(q=0.000 32 ～0.004 81,P＜0.05);而SOD值显著下降(q =0.000 18 ～0.002 53,P ＜0.05).与照射组和通气组比较,照射后通气组上述指标显著增高(q=0.004 3～0.022 6,P＜0.05);SOD值显著下降(q =0.002 9～0.008 3,P＜0.05).结论放射性肺损伤后大潮气量机械通气比正常单纯大潮气量机械通气产生了更加明显的机械通气肺损伤,表现为渗透性肺水肿、急性炎症反应、氧化应激反应的激活以及广泛的肺细胞凋亡.
목적 통과건립대서궤계통기급방사성폐손상동물모형,연구궤계통기대방사성폐손상대서폐조직급성염증、양화응격급세포조망적영향.방법 40지웅성Sprague-Dawley(SD)대서,채용수궤수자표법분성4조,매조10지,즉건강대조조、조사조、통기조화조사후통기조.관측각조폐조직병리개변;응효전영천이솔실험(EMSA)검측폐조직세포NF-κB적활성;Western blot검측폐세포핵내NF-κB아기P65단백표체수평;채용DNA단단말단표기(TUNEL)법검측폐세포적조망;동시검측폐습간중비치(W/D)、수과양화물매(MPO)、병이철(MDA)、초양화물기화매(SOD);검측폐관세액중총단백、백세포계수적수평.결과 여건강대조조상비,기여3조급성폐손상(ALI)평분、W/D비치、MPO활성、총단백、백세포계수、조망지수(AI)、폐조직MDA치、NF-κB적활성급P65단백적표체균현저성증고,차이유통계학의의(q=0.000 32 ～0.004 81,P＜0.05);이SOD치현저하강(q =0.000 18 ～0.002 53,P ＜0.05).여조사조화통기조비교,조사후통기조상술지표현저증고(q=0.004 3～0.022 6,P＜0.05);SOD치현저하강(q =0.002 9～0.008 3,P＜0.05).결론 방사성폐손상후대조기량궤계통기비정상단순대조기량궤계통기산생료경가명현적궤계통기폐손상,표현위삼투성폐수종、급성염증반응、양화응격반응적격활이급엄범적폐세포조망.
Objective To evaluate the effects of mechanical ventilation on radiation induced lung injuries of apoptosis,acute inflammation,and oxidative stress by establishing a rat mechanical ventilation model and animal model.Methods Totally 40 male Sprague-Dawley(SD) rats were randomly divided into 4 groups with 10 rats in each group:control,radiation alone,high tidal volume ventilation,and high tidal volume ventilation following by radiation.After treatment,the pathological changes in lung tissue were observed,NF-κB activity was detected by electrophoretic mobility shift assay (EMSA),the expression of NF-κB subunit P65 protein level in lung cell nucleus was detected by Western blot,and the apoptosis of lung cells was detected by terminal dexynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) method.The wet to dry weight ratio (W/D) of lung,myeloperoxidase (MPO),malondialdehyde (MDA) and superoxide dismutase (SOD) were detected.In addition,the total protein and white blood cell number in lung lavage fluid were also measured.Results Compared to the control,the acute lung injury (ALI) score,W/D ratio,MPO activity,total protein level,white blood cell number,apoptosis index (AI),lung tissue MDA,NF-κB activity and P65 protein expression were increased significantly (q =0.000 32-0.004 81,P ＜0.05),while SOD values was decreased significantly (q =0.000 18-0.002 53,P ＜0.01),in other three groups.Compared with radiation and high tidal volume ventilation group,the above indexes were significantly higher (q =0.004 3-0.022 6,P ＜ 0.05) but the SOD value was significantly lower (q =0.002 9-0.008 3,P ＜ 0.05) than those in the high tidal volume ventilation plus radiation group.Conclusions High tidal volume ventilation delivered to the radiation group produced more transparent ventilator-induced lung injury (VILI) than the high tidal volume ventilation alone induced VILI including permeable pulmonary edema,acute inflammation,oxidative stress and apoptosis in lung.