中文摘要：

目的：探讨香烟提取物（CSE）对支气管哮喘（简称哮喘）大鼠气道平滑肌细胞（ASMCs）增殖作用及可能机制。方法：16只SD大鼠随机分为对照组和哮喘组，各8只。原代培养大鼠ASMCs，取第3—6代细胞，分为对照组、对照＋CSE组、哮喘组、哮喘＋CSE组、哮喘＋CSE＋嘧啶基-苯磺酰胺（GW8510，细胞周期蛋白依赖激酶-4抑制剂）组、哮喘＋GW8510组。用流式细胞术、四甲基偶氮唑盐（MTT）法及增殖细胞核抗原（PCNA）免疫细胞化学技术检测ASMCs增殖；用逆转录-聚合酶链反应（RT—PCR）和蛋自质印迹（Western blotting）检测细胞周期蛋白D1（cyclinD1）的表达。结果：（1）哮喘组ASMCs与对照组ASMCs相比，在S＋G2／M期比例、吸光度（A）值和PCNA表达率上明显增高，差异显著（P〈0．01）。（2）哮喘组ASMCsS＋G2／M期比例、吸光度（A）值和PCNA表达率分别为（18．30±1．12）％、0．512±0．110、（55．1±3．7）％；哮喘＋CSE组分别为（32．12±1．17）％、0．801±0．210、（90．2±7．3）％；哮喘＋CSE＋GW8510组分别为（17．21±0．95）％、0．508±0．009、（54．3±4．8）％；哮喘＋GW8510组分别为（11．16±1．48）％、0．345±0．078、（40．6±5．4）％。除哮喘组、哮喘＋CSE＋GW8510组两组比较差异无显著外，其余两两比较差异均显著（P〈0．01）。（3）哮喘组、哮喘＋CSE组、哮喘＋CSE＋GW8510组、哮喘＋GW8510组ASMCs cyclin D1 mRNAA值比值和蛋白表达A值比值分别为0．236±0．045、0．271±0．002；0．369±0．124、0．379±0．002；0．231±0．075、0．261±0．002；0．165±0．064、0．193±0．002。除哮喘组、哮喘＋CSE＋GW8510组两组比较差异无显著外，其余两两比较差异均显著（P〈0．01）。结论：正常与哮喘大鼠ASMCs在CSE干预后增殖明显加快，cyclinD1表达明显增加。CSE可能是通过cyclinD1参与调控哮喘大鼠ASMCs的增殖。

英文摘要：

AIM ： To study the effect of cigarette smoke extract （CSE） on proliferation of airway smooth muscle cells （ASMCs） in bronchial asthmatic rats. METHODS： Sixteen male Sprague Dawley rats were randomly assigned into control group and asthma group. Each group had eight rats. The ASMCs in rats were cultured from primary generation. Cells between passage 3 and 6 were used in the study and were divided into control group, control ＋ CSE group, asthma group, asthma ＋ CSE group, asthma ＋ CSE ＋ GW8510 （ an inhibitor of cyclin - dependent kinase -4） group and asthma ＋ GW8510 group. The proliferation of ASMCs was examined by cell cycle analysis, MTT colorimetric assay and proliferating cell nuclear antigen （PCNA） immunocytochemical staining. The expression of cyclin D1 was detected by reverse transcriptase polymerase chain reaction （ RT - PCR） and Western blotting, respectively. RESULTS： （ 1 ） The percentage of S ＋ G2/M phase, absorbance （A） value and the expression rate of PCNA protein in ASMCs from asthma group were significantly increased compared to those in control group （P 〈0. 01 ）. （2） The percentage of S ＋ G2/M phase, A value and the expression rate of PCNA protein in ASMCs from asthma group were （18.30 ± 1.12）%, 0.512 ±0. 110 and （55.1 ± 3.7）%, respectively. Asthma ＋ CSE group were （32. 12 ± 1.17）%, 0. 801 ±0. 210 and （90. 2 ±7. 3）%, respectively. Asthma ＋ CSE ＋ GW8510 group were （ 17. 21 ± 0. 95 ） % ,0. 508 ± 0. 009 and （54. 3 ± 4. 8 ） %, respectively. Asthma ＋ GW8510 group were （ 11.16 ± 1.48 ） % , 0. 345 ± 0. 078 and （40. 6 ± 5.4） %, respectively. The significant differences between any two groups except asthma group versus asthma ＋ CSE ＋ GW8510 group （ P 〈 0. 01 ） were observed. （ 3 ） The ratios of A value of cyclin D1 mRNA and expression of cyclin D1 protein in ASMCs from asthma group, asthma ＋ CSE group, asthma ＋ CSE ＋ GW8510 group and asthma ＋ GW8510 group were 0. 236 ± 0. 045,