CAJ | 학술논문

目的探讨大鼠全肝缺血/再灌注(ischemia/reperfusion,I/R)后肺组织细胞凋亡与Bcl-XL/Bax的表达变化关系及其在肺损伤中的意义.方法 48只SD大鼠随机分成6组(n=8):①缺血前组;②冉灌注0h组;③再灌注0.5h组;④再灌注1 h组;⑤再灌注3 h组;⑥再灌注6 h组.阻断肝门30 min后开放血流,建立大鼠全肝I/R模型,48只SD大鼠分别于缺血前、再灌注0、0.5、1、3、6 h处死取肺,原位末端转移酶(terminal deoxynucleotidyl transferase dUTP nick end labeling,TUNEL)法检测细胞凋亡、PCR法检测Bcl-XL与Bax mRNA表达,同时检测肺湿/干重(W/D)比值及肺组织病理.结果 I/R后各时点肺组织W/D比值(0～6 h依次为4.96±0.25,5.30±0.32,5.36±0.32,5.71±0.33,5.32±0.28)及细胞凋亡指数[O～6 h依次为(4.86±0.64)%,(7.64±0.61)%,(15.60±0.59)%,(19.74±0.71)%,(27.48±0.88)%],与缺血前相比(分别为4.59±0.24与4.00%±0.45%)差异有统计学意义(P<0.05),并分别于再灌注3 h与6 h达峰值;Bcl-XL/Bax比值于I/R后各时点(0～6 h依次为1.01±0.13.0.43±0.03,0.51±0.07,0.62±0.06,0.62±0.07)均较缺血前(1.48±0.11)差异有统计学意义(P<0.01);病理学方面,I/R后肺泡腔完整性破坏,间隔增厚,并可见中性粒细胞浸润,这些改变于再灌注1 h已较明显,再灌注3 h最重.双变量相关分析显示,肺组织细胞凋亡指数与W/D比值呈正相关,相关系数r=0.56(P<0.01),与Bcl-XL/Bax比值呈负相关,相关系数r=0.55(P<0.01).结论大鼠全肝I/R可引起肺组织细胞凋亡加剧,并由此促进肺损伤,此过程中Bel-XL/Bax比值降低可能参与介导了细胞凋亡.
목적 탐토대서전간결혈/재관주(ischemia/reperfusion,I/R)후폐조직세포조망여Bcl-XL/Bax적표체변화관계급기재폐손상중적의의.방법 48지SD대서수궤분성6조(n=8):①결혈전조;②염관주0h조;③재관주0.5h조;④재관주1 h조;⑤재관주3 h조;⑥재관주6 h조.조단간문30 min후개방혈류,건립대서전간I/R모형,48지SD대서분별우결혈전、재관주0、0.5、1、3、6 h처사취폐,원위말단전이매(terminal deoxynucleotidyl transferase dUTP nick end labeling,TUNEL)법검측세포조망、PCR법검측Bcl-XL여Bax mRNA표체,동시검측폐습/간중(W/D)비치급폐조직병리.결과 I/R후각시점폐조직W/D비치(0～6 h의차위4.96±0.25,5.30±0.32,5.36±0.32,5.71±0.33,5.32±0.28)급세포조망지수[O～6 h의차위(4.86±0.64)%,(7.64±0.61)%,(15.60±0.59)%,(19.74±0.71)%,(27.48±0.88)%],여결혈전상비(분별위4.59±0.24여4.00%±0.45%)차이유통계학의의(P<0.05),병분별우재관주3 h여6 h체봉치;Bcl-XL/Bax비치우I/R후각시점(0～6 h의차위1.01±0.13.0.43±0.03,0.51±0.07,0.62±0.06,0.62±0.07)균교결혈전(1.48±0.11)차이유통계학의의(P<0.01);병이학방면,I/R후폐포강완정성파배,간격증후,병가견중성립세포침윤,저사개변우재관주1 h이교명현,재관주3 h최중.쌍변량상관분석현시,폐조직세포조망지수여W/D비치정정상관,상관계수r=0.56(P<0.01),여Bcl-XL/Bax비치정부상관,상관계수r=0.55(P<0.01).결론 대서전간I/R가인기폐조직세포조망가극,병유차촉진폐손상,차과정중Bel-XL/Bax비치강저가능삼여개도료세포조망.
Objective To explore the relationship of cellular apoptosis and expression of Bcl-XL/Bax in lung during total hepatic ischemia/reperfusion (I/R) in rats , and its importance in lung injury. Methods 48 healthy male SD rats were randomly divided into 6 groups(n=8): ① pre-ischemia group, ② reperfusion 0 h group, ③ reperfusion 0.5 h group, ④ reperfusion 1h group, ⑤ reperfusion 3 h group, ⑥ reperfusion 6 h group.Total hepatic I/R was developed by occlusion of hepatic helium for 30 minutes, and the occlusion was then released for reperfusion. 48 healthy male Sprague Dawley rats weighing 250 g -300 g were killed respectively prior to ischemia and at 0, 0.5, 1, 3,6 h after reperfusion, and the lung tissue was taken for determination of apoptotic cells, Bel-XL mRNA, Bax mRNA expression, wet/dry(W/D) ratio and histological examination. Results After hepatic I/R the W / D ratio (4.96±0.25, 5.30±0.32, 5.36±0.32, 5.71±0.33, 5.32±0.28)and apoptotic index (4.86±0.64)%, (7.64±0.61)%, (15.60± 0.59)%, (19.74±0.71)%and (27.48±0.88)% were increased respectively (P<0.05), compared with preischemia (4.59±0.24)% and (4.00±0.45)%. Bcl-XL/Bax ratio after hepatic I/R (1.01±0.13, 0.43±0.03, 0.51±0.07, 0.62±0.06, 0.62±0.07) were decreased (P< 0.01), compared with preischemia (1.48±0.11). Histological examination revealed that the alveolar architecture was destroyed, with interstitial thickening and neutrophil infiltration after hepatic I/R. Correlation analysis indicated that the apoptotic index showed a positive correlation with the W/D ratio (r= 0.56, P<0.01 ) and a negative correlation with the Bcl-XL/Bax ratio (r=-0.55, P<0.01). Conclusion Total hepatic ischemia-reperfusion in rats could lead to aggravation of pneumocyte apoptosis, which promoted acute lung injury. Reduction of Bcl-XL/Bax ratio during this period might mediate the occurrence of apoptosis .