CAJ | 학술논문

目的观察支气管哮喘(简称哮喘)大鼠气道重塑中气道平滑肌细胞(airway smoothmuscle cells,ASMC)凋亡及地塞米松对ASMC凋亡的影响.方法将清洁级雄性SD大鼠按随机数字表法分为正常对照组、哮喘组和地塞米松干预组,每组12只.以卵蛋白致敏和激发的方法制备大鼠慢性哮喘模型.用脱氧核糖核苷酸末端转移酶(TdT酶)介导的dUTP切口末端标记法(TUNEL法)检测ASMC凋亡并计算凋亡指数.用免疫组织化学和原位杂交法分别检测气道平滑肌Bcl-2、Bax蛋白及其mRNA的表达情况.经SPSS 11.5软件进行统计学分析,实验数据用x±s表示.多组间比较采用方差分析,多组样本均数两两比较,两变量的相关程度采用直线相关分析.结果 (1)对照组、哮喘组和干预组大鼠ASMC的凋亡指数分别为:0.201±0.022、0.030±0.016和0.118±0.043;(2)对照组、哮喘组和干预组大鼠气道平滑肌层Bcl-2蛋白表达的吸光度值分别为0.060±0.012、0.112±0.028和0.080±0.010,Bcl-2 mRNA表达的吸光度值分别为0.065±0.019、0.157±0.019和0.099±0.029;(3)对照组、哮喘组和干预组大鼠气道平滑肌层Bax蛋白表达的吸光度值为0.120±0.020、0.062±0.012和0.093±0.010,Bax mRNA表达的吸光度值分别为0.155±0.025、0.074±0.019和0.118±0.031;(4)相关分析结果显示,ASMC的凋亡指数与气道平滑肌厚度以及Bcl-2蛋白的相对含量呈显著负相关(r值分别为-0.860、-0.783,P<0.01);ASMC的凋亡指数与气道平滑肌Bax蛋白相对含量呈正相关(r=0.837,P<0.01).结论 ASMC凋亡的减少可能参与了哮喘气道重塑过程;地塞米松可以增加促凋亡蛋白Box的表达,同时减少抑凋亡蛋白Bcl-2的表达,从而增加ASMC的凋亡.
목적 관찰지기관효천(간칭효천)대서기도중소중기도평활기세포(airway smoothmuscle cells,ASMC)조망급지새미송대ASMC조망적영향.방법 장청길급웅성SD대서안수궤수자표법분위정상대조조、효천조화지새미송간예조,매조12지.이란단백치민화격발적방법제비대서만성효천모형.용탈양핵당핵감산말단전이매(TdT매)개도적dUTP절구말단표기법(TUNEL법)검측ASMC조망병계산조망지수.용면역조직화학화원위잡교법분별검측기도평활기Bcl-2、Bax단백급기mRNA적표체정황.경SPSS 11.5연건진행통계학분석,실험수거용x±s표시.다조간비교채용방차분석,다조양본균수량량비교,량변량적상관정도채용직선상관분석.결과 (1)대조조、효천조화간예조대서ASMC적조망지수분별위:0.201±0.022、0.030±0.016화0.118±0.043;(2)대조조、효천조화간예조대서기도평활기층Bcl-2단백표체적흡광도치분별위0.060±0.012、0.112±0.028화0.080±0.010,Bcl-2 mRNA표체적흡광도치분별위0.065±0.019、0.157±0.019화0.099±0.029;(3)대조조、효천조화간예조대서기도평활기층Bax단백표체적흡광도치위0.120±0.020、0.062±0.012화0.093±0.010,Bax mRNA표체적흡광도치분별위0.155±0.025、0.074±0.019화0.118±0.031;(4)상관분석결과현시,ASMC적조망지수여기도평활기후도이급Bcl-2단백적상대함량정현저부상관(r치분별위-0.860、-0.783,P<0.01);ASMC적조망지수여기도평활기Bax단백상대함량정정상관(r=0.837,P<0.01).결론 ASMC조망적감소가능삼여료효천기도중소과정;지새미송가이증가촉조망단백Box적표체,동시감소억조망단백Bcl-2적표체,종이증가ASMC적조망.
Objective To observe the changes of airway smooth muscle cells (ASMC) apoptosis in the airway remodeling process of asthma, and to evaluate the effect of dexamethasone on ASMC apoptosis and the possible mechanisms. Methods Thirty six male Sprague-Dawley rats were randomly divided into 3 groups, including a control group, an asthma group and a dexamethasone treated group. The rats were sensitized with ovalbumin and Al(OH)3, and repeatedly exposed to aerosolized ovalbumin. ASMC apoptosis was measured by the technique of TdT-mediated dUTP-biotin nick end labeling (TUNEL). Bcl-2 protein and mRNA, and Bax protein and mRNA in airway smooth muscles were measured by immunohistochemistry and in situ hybridization respectively. SPSS version 11. 5 was used for statistical analysis. Data were presented as (x±s), and means were compared with analysis of variance. The correlation of two variables was analysed by linear correlation analysis. Results The apoptosis index (AI) of ASMC was separately 0. 201±0. 022, 0. 030±0. 016, 0. 118±0. 043 in the control, the asthma, and the dexamethesone asthma,treated group. Immunohistochemistry showed that the expression of Bcl-2 protein (A) in airway smooth muscles in above groups was 0. 060±0. 012, 0. 112±0. 028 0. 080±0. 010. In situ hybridization showed that the level of Bcl-2 mRNA in airway smooth muscles in the control, the asthma, and the dexamethesone treated group was 0. 065±0. 019, 0. 157±0. 019 and 0. 099±0. 029. The expression of Bax protein in each group was 0.120±0.020, 0. 062±0.012 and 0.093±0.010 respectively. Accordingly the level of Bax mRNA in each group was 0.155±0.025, 0.074±0. 019 and 0.118±0.031 respectively. The AI of ASMC was negatively correlated with Wam/Pbm (r=-0. 860, P<0.01) and the relative content of Bcl-2 protein (r=-0.783, P<0.01), but was positively correlated with the relative content of Bax protein (r=0. 873,P <0.01). Conclusions The reduction of ASMC apoptosis may participate in the remodeling process of asthma. Dexamethasone induces ASMC apoptosis possibly by the increase of Bax expression and the decreaseof Bcl-2 expression in airway smooth muscles.