戒烟对吸烟大鼠气道上皮细胞核因子κB、基质金属蛋白酶9及金属蛋白酶组织抑制物1表达的影响

Effects of smoking cessation on expressions of nuclear factor-κB, matrix metalloproteinase-9 and tissue inhibitor of metalloproteinase-1 in airway epithelial cells of smoking rats

目的 通过研究吸烟及戒烟大鼠气道上皮细胞核因子κB（NF-κB）、细胞基质金属蛋白酶9（MMP-9）及细胞金属蛋白酶组织抑制物1（TIMP-1）mRNA及蛋白质的表达水平,探讨吸烟及戒烟对慢性阻塞性肺疾病气道炎症和气道重塑的影响.方法 雄性Wistar大鼠24只,随机分为对照组、吸烟组和戒烟组,每组8只.分别采用原位杂交和免疫组织化学的方法检测各组大鼠气道上皮细胞中NF-κB、MMP-9及TIMP-1 mRNA及蛋白的表达水平.结果 ①与对照组（0.29±0.06,0.29±0.06）相比,吸烟组（0.45±0.04,0.41±0.03）、戒烟组（0.40±0.05,0.37±0.03）NF-κB mRNA和蛋白表达水平均增高（P值均＜0.05）;与吸烟组相比,戒烟组NF-κB mRNA和蛋白表达水平降低（P值均＜0.05）.②与对照组（0.30±0.06,0.30±0.06）相比,吸烟组（0.52±0.03,0.51±0.07）、戒烟组（0.38±0.03,0.33±0.02）MMP-9 mRNA和蛋白表达水平均增高（P值均＜0.05）;与吸烟组相比,戒烟组MMP-9mRNA和蛋白表达水平均降低（P值均＜0.05）.③与对照组（0.26±0.04,0.26±0.04）相比,吸烟组（0.49±0.05,0.37±0.03）、戒烟组（0.42±0.04,0.35±0.03）TIMP-1 mRNA和蛋白表达水平均增高（P值均＜0.05）;与吸烟组相比,戒烟组TIMP-1 mRNA和蛋白表达水平降低（P值均＜0.05）.④与对照组（1.00±0.02,1.00±0.02）相比,吸烟组（1.07±0.14,l.37±0.19）MMP-9/TIMP-1 mRNA和蛋白表达值大于1（P值均＜0.05）,而戒烟组（0.92±0.13,0.94±0.10）MMP-9/TIMP-1 mRNA和蛋白表达值小于1（P值均＜0.05）.⑤各组NF-κB和MMP-9的mRNA及蛋白的表达呈正相关（r值分别为0.87和0.66,P值均＜0.05）.结论 吸烟大鼠气道上皮细胞NF-κB、MMP-9和TIMP-1 m-RNA及蛋白的表达水平升高,并且MMP-9/TIMP-1大于1,戒烟后NF-κB、MMP-9和TIMP-1的表达有所下降,且MMP-9/TIMP-1小于1,提示吸烟可以引起气道上皮的炎症和重塑,戒烟可以减轻气道炎症和重塑.

Objective To investigate the effects of smoking and smoking cessation on airway inflammation and remodeling in chronic obstructive pulmonary diseases through detecting mRNA and protein expressions of nuclear factor-κB （NF-κB）, cell matrix metalloproteinase-9 （MMP-9） and cellular tissue inhibitor of metalloproteinase-1 （TIMP-1） in airway epithelial cells of smoking and smoking cessation rats. Methods Twenty-four male Wistar rats were randomly divided into control group, smoking group and smoking cessation group,eight in each group. Hybridization in situ and immunohistochemistry were used to detect mRNA and protein expressions of NF-κB, MMP-9 and TIMP-1 in airway epithelial cells of rats. Results ① Compared with control group （0.29 ± 0.06,0.29±0.06）, mRNA and protein expressions of NF-κB in smoking group （0.45±0.04,0.41±0.03） and smoking cessation group （0.40±0.05,0.37±0.03） were higher （all P〈0.05）. The mRNA and protein expressions of NF-κB in smoking cessation group were lower than those in smoking group （all P 〈0.05）. ②Compared with control group （0.30±0.06,0.30±0.06） ,mRNA and protein expressions of MMP-9 in smoking group （0.52±0.03,0.51±0.07） and smoking cessation group （0.38±0.03,0.33±0.02） were higher （all P〈0.05）. The mRNA and protein expressions of MMP-9 in smoking cessation group were lower than those in smoking group （all P〈0.05）. ③Compared with control group （0.26±0.04, 0.26±0.04）, mRNA and protein expressions of TIMP-1 in smoking group （0.49±0.05,0.37±0.03） and smoking cessation group （0.42±0.04,0.35±0.03） were higher （all P 〈0.05）. The mRNA and protein expressions of TIMP-1 in smoking cessation group were lower than those in smoking group （all P 〈 0.05）. ④ Compared with control group （1.00±0.02,1.00±0.02）, MMP-9/TIMP-1 mRNA and protein expressions were larger than one in smoking group （1.07±0.14, 1.37±0.19）, and less than one in smoking cessation group （0.92±0.13,0.94±0.10） （all P 〈0.05）. ⑤The mRNA and protein expressions of NF-κB and MMP-9in each group were positively correlation （r=0.87,0.66,all P 〈0.05）. Conclusions In airway epithelial cells of smoking rats, mRNA and protein expressions of NF-κB, MMP-9 and TIMP-1 increase, and MMP-9/TIMP-1 is larger than one. After stoping smoking, mRNA and protein expressions of NF-κB,MMP-9 and TIMP-1 decrease, and MMP-9/TIMP-1 is less than one. This experiment explains that smoking can cause airway inflammation and remodeling, smoking cessation can reduce airway inflammation and remodeling.