摘要
目的观察大蒜素对气道平滑肌细胞（ASMC）α-平滑肌肌动蛋白（α-SMA）表达的影响，以评价大蒜素对气道重塑的抑制作用，并探讨其相应的作用机制。方法用0、2．5、5．0、10．0μmol／L的大蒜素处理人ASMC细胞48h后提取细胞总RNA和蛋白。荧光定量聚合酶链反应（PCR）检测细胞α-SMA mRNA表达，蛋白质免疫印迹法（Western blotting）检测细胞α-SMA和磷酸化Smadl（p-Smadl）的蛋白表达。结果随着大蒜素处理ASMC细胞浓度的提高（0、2．5、5．0、10．0μmol／L），细胞α-SMA mRNA表达呈进行性下降（分别为0．543±0．121、0．354±0．072、0．2234±0．058、0．191±0．034），各剂量间两两比较差异有统计学意义（均P〈0．05）；α-SMA和p-Smad1的蛋白表达也呈进行性下降[α-SMA蛋白（灰度值比值）分别为0．96±0．02、0．72±0．16、0．54±0．11、0．31±0．14，p-Smad1蛋白（灰度值比值）分别为0．94±0．03、0．76±0．13、0．62±0．11、0．43±0．12]，各剂量间两两比较差异有统计学意义（均P〈0．05）。结论大蒜素可以剂量依赖性地下调ASMC细胞α-SMA的mRNA和蛋白表达，抑制Smad蛋白的磷酸化，阻止转化生长因子-β（TGF-β）和Rho激酶信号通路。
Objective To investigate the effect of allicin on the expression of α-smooth muscle actin （ α -SMA） in airway smooth muscle cells （ASMC）, and to evaluate the mechanism of allicin on inhibition of airway remodeling. Methods The human ASMCs were treated for 48 hours with 0, 2.5, 5.0, and 10.0 μmol/L allicin, respectively, and the total RNA and protein of the cells were collected. The mRNA expression of α-SMA was determined by real-time polymerase chain reaction （PCR） analysis. The protein expression of α-SMA and phosphorylation Smadl （p-Smadl） were assessed by Western blotting analysis. Results After the treatment with allicin for 48 hours in a dose of 0, 2.5, 5.0, and 10.0μmol/L respectively, the mRNA expression of α-SMA was down-regulated （0.543 ± 0.121, 0.354 ± 0.072, 0.223 ± 0.058, and 0.191 ± 0.034, respectively）, with statistically significant difference among groups （all P〈0.05）, and the protein expression of α-SMA and p-Smadl was also gradually down-regulated in a dose-dependent manner [α -SMA protein （grad value ratio） ： 0.96 ± 0.02, 0.72 ± 0.16, 0.54 ±0.11, and 0.31 ±0.14, respectively; p-Smadl protein （grad value ratio）： 0.94 20.03, 0.76 ±0.13, 0.62 , 0. 11, and 0.43 ± 0.12, respectively）, with statistically significant difference among groups （all P〈0.05 ）. Conclusion Allicin depresses the mRNA and protein expression of α-SMA, and inhibits p-Smadl in a dose-dependent manner, thus interrupts the transforming growth factor- β and Rho kinase signal pathway.
出处
《中华危重病急救医学》
CAS
CSCD
北大核心
2013年第3期164-166,共3页
Chinese Critical Care Medicine
基金
基金项目：吴阶平医学基金会临床科研专项基金（32067501083）
