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The effect of cigarette smoke extract on the proliferation of human airway smooth muscle cells sensitized by serum from bronchial asthmatic patients

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Author:
No author available
Journal Title:
CHINESE JOURNAL OF TUBERCULOSIS AND RESPIRATORY DISEASES
Issue:
8
DOI:
10.3760/cma.j.issn.1001-0939.2011.08.018
Key Word:
烟雾;哮喘;肌细胞,平滑肌;增殖;细胞周期蛋白D1;Smog;Asthma;Myocytes,smooth muscle;Proliferation;Cyclin D1

Abstract: Objective To study the effect of cigarette smoke extract (CSE) on the proliferation of human airway smooth muscle cells (HASMCs) sensitized by serum from asthmatic patients and the underlying mechanisms.Methods HASMCs were cultured from primary generation.Cells between passage 4 and 8 were used in the study.HASMCs were sensitized by 10% serum from asthmatic patients and were divided into an asthmatic serum group, an asthmatic serum + CSE group, an asthmatic serum + GW8510 (inhibitor of cyclin-dependent kinase-4) group and an asthmatic serum + CSE + GW8510 group.Non-asthmatic human serum treated HASMCs served as the control.The proliferation of HASMCs was examined by cell cycle analysis, MTT colorimetric assay and[3H]thymidine incorporation.The expression of cyclinD1 was detected by reverse transcriptase polymerase chain reaction (RT-PCR) and Western blotting.Results The percentage of S + G2/M phase, the absorbance (A) value and the DNA synthesis value in asthmatic serum group were significantly increased compared with those of the control group ( q = 6.25,5.61,6.82,respectively, all P <0.01 ).The percentage of S + G2/M phase, the absorbance (A) value and DNA synthesis value in the asthmatic serum group were (21.4 ± 1.1 ) %, 0.392 ± 0.124 and 2669 ± 138,respectively.Their value in the asthmatic serum + CSE group were (33.3 ± 1.3)%, 0.612 ±0.201 and 3552 ± 303, respectively, which were significantly increased compared with those of the asthmatic serum group( q = 5.67,6.32,5.56, respectively, all P < 0.01 ).Their value in the asthmatic serum + GW8510 group were (14.7 ± 1.4)%, 0.301 ±0.097 and 1812 ± 109, respectively, which were significantly decreased compared with those of the asthmatic serum group ( q = 6.02,5.53,5.79, respectively, all P <0.01 ).The ratios of A value of cyclinD1 mRNA and the expression of cyclinD1 protein in the asthmatic serum group were 0.291 ±0.112 and 0.186 ±0.002, respectively.The ratios of A value in the asthmatic serum + CSE group were 0.521 ± 0.102 and 0.312 ± 0.002, respectively, which were significantly increased compared with those of the asthmatic serum group (q = 12.09,9.26, respectively, all P < 0.01 ).The ratios of A value in the asthmatic serum + GW8510 group were 0.223 ±0.038 and 0.150 ±0.002,respectively, which were significantly decreased compared with those of the asthmatic serum group (q =6.86,5.60, respectively, all P < 0.01 ).Conclusions HASMCs sensitized by serum from asthmatic patients showed accelerated proliferation after intervention by CSE, with increased expression of cyclinD1.CSE may increase the proliferation of HASMCs sensitized by serum from asthmatic patients via regulating cyclinD expression.

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