摘要
目的:本研究主要探讨了肿节风对小型猪经右胸单次照射后上皮间质转化(EMT)的抑制作用及其分子机制。方法:雄性小型猪随机分为对照组、单照组和药照组,单照组和药照组行右胸单次15 Gy照射,对照组不予照射。药照组于照射前一周予肿节风配方颗粒溶液,对照组和单照组予等量生理盐水。照射后不同时间点分别从三组随机取5头小型猪采集右肺组织,行免疫印迹检测肺表面活性蛋白A(SP-A)、α平滑肌肌动蛋白(α-SMA)、vimentin和E-cadherin的蛋白表达;免疫荧光双染观察α-SMA和SP-A的共表达。结果:在照射后不同时间点,单照组中α-SMA和vimentin蛋白表达较对照组明显升高;而SP-A和E-cadherin蛋白表达却呈下降趋势,药照组中,上述4个观察指标的蛋白表达则介于二者之间,且4个不同时间点,三组之间两两比较,差异均具有统计学意义。单照组和药照组在激光共聚焦显微镜下均观察到了肺泡II型上皮细胞出现α-SMA和SP-A共染,对照组则没有此现象。结论:肿节风可能通过抑制肺泡II型上皮细胞发生EMT来减少胶原纤维的产生,进而减慢放射性肺纤维化的进展。
This study aimd to investigate the inhibitory effects and molecular mechanism of sarcandra glabra on epithelial-mesenchymal transition（EMT） produced by right thoracic single irradiation of minipigs. Minipigs were randomly divided into the control group, the radiotherapy group and the medicine plus radiotherapy group. Minipigs of the radiotherapy group and the medicine plus radiotherapy group received either right thoracic irradiation（15 Gy） or sham-irradiation; while those of the medicine plus radiotherapy group took the prescription of sarcandra glabra orally one week prior to the radiotherapy. Meanwhile, minipigs of the control and the radiotherapy groups were administered with the same volume of physiological saline as the prescription. Finally, five minipigs were randomly selected from each group, and right lung tissue was prepared. Thus, the protein levels of SP-A, α-SMA, vimentin and E-cadherin were detected by western blot, while the distributions of SP-A and α-SMA were merged via double immunofluorescence staining. It was found that the protein expressions of α-SMA and vimentin increased, while those of SP-A and E-cadherin decreased in the radiotherapy group compared with the control group within a time-dependent manner postirradiation. The protein levels of α-SMA, vimentin, SP-A and E-cadherin in the medicine plus radiotherapy group were in the midst of the expression ranges of the control and the radiotherapy groups, which showed significant statistic differences according to the parewise comparison method. Moreover, the coexpression of SP-A and α-SMA was observed in type II alveolar epithelial cells in the radiotherapy group and the medicine plus radiotherapy group via a confocal laser scanning microscope, while the coexpression didn’t presented in the control group. In conclusion, a lower level of collagen fibers may be synthesized out of suppressing the EMT of type II alveolar epithelial cells and the slowdown in the process of radioactive pulmonary fibrosis.
出处
《世界科学技术-中医药现代化》
2016年第5期846-853,共8页
Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology
基金
广西壮族自治区科学技术厅科技攻关与新产品试制项目(1140003A-24):中草药对放射性肺损伤的防护研究
负责人:刘文其
关键词
放射治疗
小型猪
肿节风
肺泡Ⅱ型上皮细胞
上皮间质转化
Radiotherapy
minipigs
sarcandra glabra
type II alveolar epithelial cells
epithelial-mesenchymal transition
