CAJ | 학술논문

目的探讨幼年哮喘气道重塑大鼠中气管平滑肌细胞骨架的变化及RhoA/ROCK信号通路的影响.方法应用抗原致敏液对幼年SD大鼠激发8周,原代培养纯化气管平滑肌细胞(ASMC),并给予Rho激酶抑制剂Y27632干预,通过免疫荧光、实时定量PCR、Western blot等方法,检测气管平滑肌细胞骨架F-actin、α-tubulin的变化.结果 (1)哮喘气道重塑组ASMC中F-actin平均灰度值高于对照组(P＜0.01),加入Y27632预处理后,F-actin平均灰度值明显降低(P＜0.01);(2)哮喘气道重塑组α-tubulin的荧光信号强度、密度较对照组增高,而Y27632干预组微管α-tubulin的荧光信号明显减弱(P＜0.05);(3)哮喘气道重塑组α-tubulin蛋白的表达较对照组显著增加(P＜0.01),Y27632预处理组α-tubulin蛋白的表达明显下降(P＜0.05).结论幼年哮喘气道重塑大鼠中存在着气管平滑肌细胞骨架的变化,RhoA/ROCK信号通路可能起着重要作用.
목적 탐토유년효천기도중소대서중기관평활기세포골가적변화급RhoA/ROCK신호통로적영향.방법 응용항원치민액대유년SD대서격발8주,원대배양순화기관평활기세포(ASMC),병급여Rho격매억제제Y27632간예,통과면역형광、실시정량PCR、Western blot등방법,검측기관평활기세포골가F-actin、α-tubulin적변화.결과 (1)효천기도중소조ASMC중F-actin평균회도치고우대조조(P＜0.01),가입Y27632예처리후,F-actin평균회도치명현강저(P＜0.01);(2)효천기도중소조α-tubulin적형광신호강도、밀도교대조조증고,이Y27632간예조미관α-tubulin적형광신호명현감약(P＜0.05);(3)효천기도중소조α-tubulin단백적표체교대조조현저증가(P＜0.01),Y27632예처리조α-tubulin단백적표체명현하강(P＜0.05).결론 유년효천기도중소대서중존재착기관평활기세포골가적변화,RhoA/ROCK신호통로가능기착중요작용.
Objective To investigate the alteration of the cytoskeleton of airway smooth muscle cells in young asthmatic rats with airway remodeling and the effect of RhoA/ROCK signal pathway.Methods Airway smooth muscle cells (ASMCs) were primary cultured and purified from Sprague-Dawley(SD) rats that were induced by ovalbumin (OVA) inhalation for 8w,then incubated by Pho kinase inhibitor Y27632.Real time quantitative polymerase chain reaction (RT-PCR),Western blot,and immunohistochemistry were used to measure the alteration of F-actin,and α-tubulin in the cytoskeleton of airway smooth muscle.Results (1) The asthma group showed a high average gray value of F-actin in ASMC than control groups,especially 8 weeks;and were significantly down in the group after adding Y27632(P ＜0.01).(2) The intension and intensity of fluorescence signal of α-tubulin in asthma groups in 8 weeks were higher than control greup(P ＜0.01),and were significantly decreased in Y27632 group.(3) A higher expression of α-tubulin protein was shown in the asthma group in 8 weeks relative to control group(P ＜0.01),and was significantly down-regulated in Y27632 group(P ＜0.05).Conclusions Alteration of the cytoskeleton of airway smooth muscle exists in young asthmatic rats and the RhoA/ROCK signal pathway possibly plays a significant role.