摘要
目的探讨罗红霉素(RXM)经caveolin-1-p-ERK1/2对离体哮喘大鼠气道平滑肌细胞(ASMCs)cyclinD1表达的影响。方法将30只SPF级雄性SD大鼠随机均分为对照组和哮喘组,以卵白蛋白致敏和激发复制哮喘气道重塑模型;采用Image-Pro Plus Version 6.0图像分析软件测定支气管壁和支气管平滑肌层厚度;透射电镜观察两组大鼠ASMCs微囊表达情况;CCK-8检测不同浓度RXM[A组(只含0.1%DMSO)、R10组(含RXM10μg/ml+0.1%DMSO)、R25组(含RXM 25μg/ml+0.1%DMSO)、R50组(含RXM50μg/ml+0.1%DMSO)、R100组(含RXM 100μg/ml+0.1%DMSO)]干预哮喘ASMCs后细胞的增殖情况。Western blot测定caveolin-1、p-ERK、cyclinD1的蛋白表达。结果哮喘组支气管壁和支气管平滑肌层明显增厚。而微囊表达匮乏。哮喘组大鼠支气管壁与平滑肌层厚度均高于对照组(均P<0.01)。各RXM干预组ASMCs A值均低于A组。其中R100组明显低于A组(P<0.05)。R100组caveolin-1表达明显高于A组及R10组,cyclinD1表达明显低于A组及R10组,差异均有统计学意义(P<0.05或0.01);R25、R50、R100组P-ERK表达均明显低于A组.差异均有统计学意义(P<0.05或0.01)。哮喘ASMCs增殖活性与caveolin-1蛋白表达呈负相关(P<0.05),与P-ERK及cyclinD1蛋白表达呈正相关(P<0.05或0.01);caveolin-1与p-ERK蛋白表达呈负相关(P<0.01),与cyclinD1蛋白表达呈正相关(P<0.01);P-ERK与cyclinD1蛋白表达呈正相关(P<0.01)。结论 RXM可能通过上调caveolin-1表达、抑制ERK1/2活化。进而影响cyclinD1的表达.从而抑制哮喘大鼠ASMCs增殖。
Objective To investigate the effect of roxithromycin （RXM） on cyclinD1 expression in cultured airway smooth muscle cells （ASMCs）of asthmatic rats and its mechanism. Methods Thirty male Sprague-Dawtey rats were randomly divided into control group and asthmatic group. The chronic asthma was induced by the sensitization and provocation of ovalbumin in rats. The total bronchial wall thickness（Wat/Pbm） and the bronchial smooth muscle thickness（Wam/Pbm） were measured with im- age-pro plus 6.0;the changes of caveotae in AMSCs were observed by transmission electron microscope. The proliferation of cultured ASMCs induced by different concentrations of roxithromycin（group A： 0.1% DMSO, group R10： RXM lOiJg/mL＋0.1%DM- SO, groupR25： RXM 25ug/ml＋0.1% DMSO, group R50： RXM501Jg/mI＋0.1%DMSO, group R 100： RXM 1001Jg/ml＋0.1% DMSO was tested by CCK-8. The protein expressions of caveolin-1, P-ERK, cyclinD1 of ASMCs were measured by Western blot. Results The Wat/Pbm, Warn/Pbm of asthmatic group were significantly higher than those of control group （P〈0.01）. There were a few of caveolae in ASMCs of asthmatic rats. A value in all intervention groups was lower than that in group A, particularly in group RIO0 （P〈0.05）. The expression of caveolin-1 in group RIO0 was significantly higher and the expression of cyclinD1 was significantly lower than those in group A and group R10 （P〈O.05 or P〈O.01）; the expression of P-ERK in group R25, R50, RIO0 were signifi- cantly lower than those in group A （P〈0.05 or P〈0.01）. There were significantly negative correlation of cell activity rate of asth- matic rats with the expressions of caveolin-1 protein （P〈0.05） and positive correlation with the expressions of P-ERK or cyclinD1 protein （P〈0.05 or P〈0.01）. The expression of caveolin-1 protein was negatively correlated with P-ERK and cyclinD1 （P〈0.01）, and the expression of P-ERK protein was positively correlated with eyclinD1 （P〈0.01）. Conclusion Roxithromycin inhib
出处
《浙江医学》
CAS
2014年第5期366-369,397,共5页
Zhejiang Medical Journal
基金
浙江省卫生厅科研基金资助项目(2009A144)
