中华结核和呼吸杂志
中華結覈和呼吸雜誌
중화결핵화호흡잡지
Chinese Journal of Tuberculosis and Respiratory Diseases
2011年
8期
604-608
,共5页
张晓宇%马利军%徐永健%刘先胜%张珍祥
張曉宇%馬利軍%徐永健%劉先勝%張珍祥
장효우%마리군%서영건%류선성%장진상
烟雾%哮喘%肌细胞,平滑肌%增殖%细胞周期蛋白D1
煙霧%哮喘%肌細胞,平滑肌%增殖%細胞週期蛋白D1
연무%효천%기세포,평활기%증식%세포주기단백D1
Smog%Asthma%Myocytes,smooth muscle%Proliferation%Cyclin D1
目的 探讨香烟烟雾提取物(CSE)对支气管哮喘(简称哮喘)患者血清致敏的人气道平滑肌细胞(HASMC)增殖作用及其可能机制.方法 原代培养HASMC,取第4～8代细胞,用10%哮喘患者血清致敏HASMC,分为血清组、血清+CSE组、血清+GW8510(嘧啶基-苯磺酰胺,细胞周期蛋白依赖激酶-4抑制剂)组、血清+CSE+GW8510组,以10%非哮喘患者血清为对照组.用流式细胞术、四甲基偶氮唑盐(MTT)法及3H-TdR掺入法检测HASMC增殖,用逆转录-聚合酶链反应(RT-PCR)和Western blot检测细胞周期蛋白D1(cyclinD1])的表达.结果 血清组HASMC的S+G2 M期比例、吸光度(A)值和细胞DNA合成量[分别为(21.4±1.1)%、0.39±0.12和2669±138]明显高于对照组,差异均有统计学意义(q值分别为6.25、5.61、6.82,均P＜0.01);血清+CSE组HASMC的S+G2M期比例、A值和DNA合成量分别为(33.3±1.3)%、0.61±0.20和3552±303,与血清组比较差异均有统计学意义(q值分别为5.67、6.32、5.56,均P＜0.01);血清+GW8510组HASMC的S+G2M期比例、A值和DNA合成量分别为(14.7±1.4)%、0.30±0.10和1812±109,与血清组比较差异均有统计学意义(q值分别为6.02、5.53、5.79,均P＜0.01).血清+CSE组HASMC cyclinD1 mRNAA值比和蛋白表达A值比分别为0.521±0.102和0.312±0.002,与血清组(分别为0.291±0.112和0.186±0.002)比较,差异均有统计学意义(q值分别为12.02和9.26,均P＜0.01);血清+GW8510组分别为0.223±0.038、0.150±0.002,与血清组比较差异均有统计学意义(q值分别为6.86、5.60,均P＜0.01).结论 CSE可能通过cyclinD参与调控哮喘患者血清致敏的HASMC增殖.
目的 探討香煙煙霧提取物(CSE)對支氣管哮喘(簡稱哮喘)患者血清緻敏的人氣道平滑肌細胞(HASMC)增殖作用及其可能機製.方法 原代培養HASMC,取第4～8代細胞,用10%哮喘患者血清緻敏HASMC,分為血清組、血清+CSE組、血清+GW8510(嘧啶基-苯磺酰胺,細胞週期蛋白依賴激酶-4抑製劑)組、血清+CSE+GW8510組,以10%非哮喘患者血清為對照組.用流式細胞術、四甲基偶氮唑鹽(MTT)法及3H-TdR摻入法檢測HASMC增殖,用逆轉錄-聚閤酶鏈反應(RT-PCR)和Western blot檢測細胞週期蛋白D1(cyclinD1])的錶達.結果 血清組HASMC的S+G2 M期比例、吸光度(A)值和細胞DNA閤成量[分彆為(21.4±1.1)%、0.39±0.12和2669±138]明顯高于對照組,差異均有統計學意義(q值分彆為6.25、5.61、6.82,均P＜0.01);血清+CSE組HASMC的S+G2M期比例、A值和DNA閤成量分彆為(33.3±1.3)%、0.61±0.20和3552±303,與血清組比較差異均有統計學意義(q值分彆為5.67、6.32、5.56,均P＜0.01);血清+GW8510組HASMC的S+G2M期比例、A值和DNA閤成量分彆為(14.7±1.4)%、0.30±0.10和1812±109,與血清組比較差異均有統計學意義(q值分彆為6.02、5.53、5.79,均P＜0.01).血清+CSE組HASMC cyclinD1 mRNAA值比和蛋白錶達A值比分彆為0.521±0.102和0.312±0.002,與血清組(分彆為0.291±0.112和0.186±0.002)比較,差異均有統計學意義(q值分彆為12.02和9.26,均P＜0.01);血清+GW8510組分彆為0.223±0.038、0.150±0.002,與血清組比較差異均有統計學意義(q值分彆為6.86、5.60,均P＜0.01).結論 CSE可能通過cyclinD參與調控哮喘患者血清緻敏的HASMC增殖.
목적 탐토향연연무제취물(CSE)대지기관효천(간칭효천)환자혈청치민적인기도평활기세포(HASMC)증식작용급기가능궤제.방법 원대배양HASMC,취제4～8대세포,용10%효천환자혈청치민HASMC,분위혈청조、혈청+CSE조、혈청+GW8510(밀정기-분광선알,세포주기단백의뢰격매-4억제제)조、혈청+CSE+GW8510조,이10%비효천환자혈청위대조조.용류식세포술、사갑기우담서염(MTT)법급3H-TdR참입법검측HASMC증식,용역전록-취합매련반응(RT-PCR)화Western blot검측세포주기단백D1(cyclinD1])적표체.결과 혈청조HASMC적S+G2 M기비례、흡광도(A)치화세포DNA합성량[분별위(21.4±1.1)%、0.39±0.12화2669±138]명현고우대조조,차이균유통계학의의(q치분별위6.25、5.61、6.82,균P＜0.01);혈청+CSE조HASMC적S+G2M기비례、A치화DNA합성량분별위(33.3±1.3)%、0.61±0.20화3552±303,여혈청조비교차이균유통계학의의(q치분별위5.67、6.32、5.56,균P＜0.01);혈청+GW8510조HASMC적S+G2M기비례、A치화DNA합성량분별위(14.7±1.4)%、0.30±0.10화1812±109,여혈청조비교차이균유통계학의의(q치분별위6.02、5.53、5.79,균P＜0.01).혈청+CSE조HASMC cyclinD1 mRNAA치비화단백표체A치비분별위0.521±0.102화0.312±0.002,여혈청조(분별위0.291±0.112화0.186±0.002)비교,차이균유통계학의의(q치분별위12.02화9.26,균P＜0.01);혈청+GW8510조분별위0.223±0.038、0.150±0.002,여혈청조비교차이균유통계학의의(q치분별위6.86、5.60,균P＜0.01).결론 CSE가능통과cyclinD삼여조공효천환자혈청치민적HASMC증식.
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.
