香烟烟雾暴露对大鼠支气管肺泡灌洗液及外周血4-HNE和γ-GCS表达的影响

Effect of Cigarette Smoke Exposure on the Expression of 4-HNE and γ-GCS in Bronchoalveolar Lavage Fluid(BALF) and Peripheral Blood of Rats

目的研究香烟烟雾暴露对大鼠支气管肺泡灌洗液(BALF)及外周血4-羟基壬稀醛(4-hydroxynonenal,4-HNE)和γ-谷氨酰半胱氨酸合成酶(gamma-Glutamylcysteine synthetase,γ-GCS)表达的影响,探讨4-HNE及γ-GCS在COPD氧化$抗氧化失衡发病机制中的作用。方法 30只雄性Wistar大鼠随机分为不吸烟组、吸烟6周组和吸烟12周组;采用双抗体夹心酶联免疫吸附(ABC-ELISA)法分别检测各组大鼠BALF及外周血4-HNE和γ-GCS的含量。结果 (1)与不吸烟组(15.917±1.584;12.305±1.822)比较,吸烟6周组(21.376±2.80;18.333±2.585)和吸烟12周组(28.634±2.999;22.754±3.882)大鼠BALF和外周血中4-HNE含量明显增加,差异均有统计学意义(P均<0.01);吸烟12周组与吸烟6周组比较有所增加,差异也有统计学意义(P<0.01)。(2)与不吸烟组(13.525±2.311;11.847±1.171)比较,吸烟6周组(15.802±1.647;16.221±1.577)大鼠BALF和外周血中γ-GCS的表达明显增加,差异均有统计学意义(P<0.05;P<0.01);与不吸烟组比较,吸烟12周组(11.527±1.606;9.015±1.569)的表达明显减少,差异也有统计学意义(P<0.05;P<0.01)。(3)吸烟6周组BALF及外周血4-HNE和γ-GCS的表达呈正相关(r=0.764,r=0.674),吸烟12周组BALF中4-HNE和γ-GCS表达呈负相关(r=-0.777),差异均有统计学意义(P均<0.05),而外周血中二者表达无显著相关性。结论香烟烟雾暴露可使大鼠体液4-HNE及γ-GCS的表达水平增高,随着吸烟时间延长,γ-GCS逐渐降低,提示二者在COPD氧化$抗氧化失衡中发挥一定的作用。

Objective To explore the effect of cigarette smoke exposure on the expression of 4-hydroxynonenal（4-HNE） and gamma-glutamylcysteine synthetase（γ-GCS）in bronchoalveolar lavage fluid（BALF） and peripheral blood of rats.Methods Thirty male Wistar rats were randomly divided into three groups： non-smoking group,6-weeks smoking group and 12-weeks smoking group.4-HNE and γ-GCS in BALF and peripheral blood were quantitatively analyzed by avidin-biotin complex enzyme-linked immunosorbent assay（ABC-ELISA）.Results（1）The expressions of 4-HNE in BALF and peripheral blood of rats in the 6-weeks smoking group（21.376±2.80;18.333±2.585）and the 12-weeks smoking group（28.634±2.999;22.754±3.882）were significantly higher than those in the non-smoking group（15.917±1.584;12.305±1.822）（all P0.01）,and those of the 12-weeks smoking group were significantly higher than those of the 6-weeks smoking group（all P0.01）.（2）The expression of γ-GCS in BALF and peripheral blood of rats in the 6-weeks smoking group（15.802±1.646;16.221±1.577）were significantly higher than those in the non-smoking group（13.525±2.311;11.847±1.171）（P0.05;P0.01）,and those of the 12-weeks smoking group（11.527±1.606;9.015±1.569）were significantly lower than those of the non-smoking group（P0.05;P0.01）.（3）The level of 4-HNE in BALF and peripheral blood in the 6-weeks smoking group was positively related with that of γ-GCS（r=0.764,r=0.674）,and the level of 4-HNE in BALF of the 12-weeks smoking group was negatively related with γ-GCS（r=-0.777）（all P0.05）,but there was no significant correlation between the 4-HNE and γ-GCS in peripheral blood of the 12-weeks smoking group.Conclusions Cigarette smoke exposure can upregulate the expression of 4-HNE and γ-GCS in BALF and peripheral blood in rats.As smoking is prolonged,the expression of γ-GCS is gradually reduced,which indicates that 4-HNE and γ-GCS may play an important role in the oxidation/antioxidant imbalance in chronic obstructive pulmonary disease.