摘要
目的探讨微囊蛋白1抑制气道平滑肌细胞（ASMCs）增殖的可能机制以及罗红霉素的调控作用。方法复制哮喘大鼠模型，光镜观察肺组织病理学变化，透射电镜观察各组ASMCs上微囊的结构及变化，用组织贴壁法体外培养ASMCs，实验设对照组（A组：含2．5％胎牛血清的高糖培养液处理1h）、哮喘组（B组：处理同A组）、细胞外调节蛋白激酶（ERK1／2）信号通路阻断剂PD98059组（c组：10×10μmol／L的PD98059处理1h）、罗红霉素组（D组：100mg／L的罗红霉素处理1h）、β-甲基环糊精组（E组：5μmoL／L的B甲基环糊精处理1h）；用细胞计数试剂盒（CCK-8）检测各组细胞的增殖情况，Western印迹检测各组微囊蛋白1表达；逆转录（RT）-PCR和Western印迹分别检测ERK1／2mRNA、单核细胞趋化蛋白1（MCP-1）mRNA和磷酸化ERK1／2、MCP-1蛋白的表达。结果c、D组ASMCs增殖显著低于B组（0．68±0．15、0．63±0．13比0．96±0．14，均P〈0．05）；E组（1．26±0．11）ASMCs增殖强于B组（P〈0．05）。C、D组微囊蛋白1表达均显著高于B组（0．332±0．057、0．392±0．064比0．237±0．032，均P〈0．05）；C、D组ERKl／2蛋白表达均显著低于B组（0．241±0．017、0．268±0．007比0．346±0．009，均P〈0．01），E组（0．441±0．011）ERK1／2蛋白表达高于B组；C、D组ERKl／2mRNA表达均显著低于B组（0．277±0．043、0．338±0．026比0．591±0．022，均P〈0．01）。C、D组MCP-1蛋白表达均显著低于B组（0．198±0．015、0．286±0．019比0．482±0．026，均P〈0．01），E组（0．521±0．023）MCP-1蛋白表达较B有升高趋势；C、D组MCP-1mRNA表达均显著低于B组（0．212±0．042、0．249±0．032比0．676±0．053，均P〈0．01）。结论微囊蛋白1可能通过ERK信号通路抑制MCP-1表达，从而抑制哮喘ASMCs的增殖。罗红霉素可能上调微囊蛋白1的表达，抑制MCP-1mRNA表达和翻译，进而抑制哮喘ASMCs的增殖。
Objective To explore the functional role of caveolin-1 in airway smooth muscle cells （ASMCs） proliferation and examine the regulatory effect of roxithromycin. Methods The rat model of bronchial asthma was established. Electron microscope was employed to observe the status of caveolae and light microscope for the histological changes in pulmonary tissues. The primarily cultured ASMCs were divided into 5 groups： control （ group A）, asthmatic ASMCs （ group B）, PD98059 （ group C）, roxithroymcin （group D） and methyl-[5-cyclodextrin （group E）. Cell proliferation was detected by Cell Counting Kit-8 （CCK-8）. And the expressions of caveolin-1, extracellular regulated protein kinases （ERK） and monocyte chemotactic protein （MCP）-I were detected by Western blot and reverse transcription polymerase chain reaction （RT-PCR）. Results The cell proliferation of asthmatic ASMCs （0. 68±0.15, 0. 63±0. 13） in groups C and D were significantly less than those in group B （0. 96±0. 14） （both P 〈 0. 05） while group E was more than group B （ 1.26 ±0. 11 vs 0. 96± 0. 14, P 〈 0. 05 ）. The content of caveolin-1 （0. 392±0. 064, 0. 332 ±0. 057） in groups C and D were higher than those in group B （0. 237 ±0. 032） （both P 〈0. 05） while ERK1/2 protein level in groups C and D （0. 241±0. 017, 0. 268±0. 007） were less than those in group B （0. 346±0. 009） （both P 〈0. 01 ）. And MCP-1 protein level in groups C and D （0. 198 ±0.015, 0.286 ±0.019） were less than those in group B （0.482±0.026） （both P 〈0. 01 ）. The ERK mRNA level in groups C and D （0. 277 ±0. 043, 0. 338 ±0. 026） were less than those in group B （0. 591±0. 022） （both P 〈 0. 01 ）. And also MCP-1 mRNA in groups C and D （0. 212 ±0. 042, 0.249±0.032） were less than those in group B （0.676±0.053） （all P 〈 0.01）. Conclusions Caveolin-1 preventing the proliferation of asthmatic ASMCs is most likely mediated by ERK1/2 signal pathway and a down-regulation of MCP-1 expression. And roxithroymein reduces the proliferation of asthmatic ASMCs through up-regulating the expression of eaveolin-1 and inhibiting the expression of MCP-1.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2013年第34期2750-2754,共5页
National Medical Journal of China
基金
基金项目：浙江省自然科学基金（Y2080466）
浙江省医药卫生科技计划（2009A144）
