caveolin-1蛋白及ERK1/2信号途径在罗红霉素抑制哮喘气道平滑肌细胞增殖中的作用

The effects of caveolin-1 protein and ERK1/2 signal pathway on the inhibition of the proliferation of the asthmatic airway smooth muscle cells induced by roxithromycin

目的 :研究微囊蛋白-1(caveolin-1)及ERK1/2信号通路在罗红霉素抑制哮喘大鼠气道平滑肌细胞(ASMCs)增殖中的作用。方法:培养正常大鼠及哮喘模型大鼠ASMCs。哮喘组细胞以罗红霉素、甲基-β-环糊精、ERK1/2信号通路阻断剂PD98059分别进行干预。用CCK-8法检测各组细胞的增殖情况,Westernblot(WB)法检测各组细胞caveolin-1蛋白、磷酸化ERK1/2(pERK1/2)蛋白、细胞周期蛋白D1(cyclinD1)表达情况。结果:CCK-8检测结果显示,罗红霉素组(0.6807±0.15374)与哮喘组(0.9592±0.13728)比较,OD值降低,P<0.05;PD98059组(0.7567±0.16167)与哮喘组(0.9592±0.13728)比较,OD值亦降低,P<0.05;甲基-β-环糊精组(1.2583±0.20494)OD值较哮喘组(0.9592±0.13728)高,P<0.05。WB法检测结果显示,体外培养的哮喘大鼠ASMCs经罗红霉素干预后caveolin-1蛋白表达量较哮喘组升高,ERK1/2的活化型降低,cyclinD1减少;甲基-β-环糊精组则相反。应用ERK1/2的阻断剂PD98059作用于体外培养的哮喘大鼠ASMCs,结果ERK1/2的活化型pERK表达量明显降低,cyclinD1减少。结论:罗红霉素可以上调caveolin-1蛋白的表达量,抑制ERK1/2途径,使cyclinD1的表达量减少,从而抑制哮喘大鼠ASMCs的增殖。

Objective： To study the role of caveolin-I protein and ERK1/2 signal pathway in the inhibition of the asthmatic airway smooth muscule cells induced by roxithromycin. Methods： ASMCs was cultured in medium of 10% fetal bovine serum （FBS）. Then roxithroymcin （RXM）, β-cyclodextrin, MAPKs signal pathway＇s inhabitor PD98059 were added to the medium to culture the ASMCs respectively. Cell proliferations of each group were assessed using CCK-8. Western blot was used to detect the expression of caveolin-1, pERK and cyclinD1 protein. Results： The values of OD in the RXM group （0.6807 ± 0.15374） and PD98059 group （0.7567 ± 0.16167） were lower than that in the asthmatic group （0.9592 ± 0.13728）, the value of OD in β - cyclodextrin group （1.2583 ± 0.20494） was higher than that in the asthmatic group. Western blot showed the increased expression of caveolin-1, decreased expression of p-ERK1/2 and cyclinD 1 in RXM group, β- cyclodextrin could reverse it. MAPK inhibitor PD98059 could reduce the expressions of p-ERK and cyclinD 1 proteins. Conculusion： RXM can decrease the expression of cyclinD1 by supressing the expression of ERK signal pathway, which is regulated by the up-regulated level of caveolin-1 protein, resulting in the inhibition of the asthmatic ASMCs. ERK signal pathway is involved in the proliferation of the asthmatic ASMCs.