摘要
目的体外观察炎调方通过Fas/FasL通路延缓LPS诱导II型肺泡上皮细胞(ATII)凋亡的作用。方法原代分离与培养ATII细胞,运用脂多糖(LPS)干预ATII进行体外ALI细胞造模,结合药物血清技术,观察炎调方对ATII细胞凋亡的影响。Hoechst 33258荧光染色观察细胞凋亡形态,流式细胞仪检测ATII细胞凋亡率,Western blot方法检测Fas、FasL、caspase-8、caspase-3表达。结果 LPS降低了ATII细胞存活率,炎调方对LPS诱导的细胞凋亡有显著的抑制作用;LPS能明显激活caspase-8和caspase-3活性,提高Fas和FasL的水平;炎调方能减弱caspase-8和caspase-3活性,降低Fas和FasL表达。结论炎调方能延缓ATII凋亡,其机制可能与抑制Fas/FasL通路活性有关。
Objective To observe the effect of Yantiao Fang on ATII apoptosis through Fas/FasL pathway in vitro. Methods II type alveolar epithelial cells（ATII） cells are primary isolated and cultured. ATII cells are treated with LPS for the modeling of ALI cells in vitro. With the application of drug serum technique, the effect of Yantiao Fang on apoptosis of ATII cells is observed. Hoechst 33258 fluorescence staining is used to observe the morphology of cell apoptosis. Flow cytometry is used to detect apoptosis rate of apoptotic cells in ATII cells. The expressions of Fas, FasL, caspase-8 and caspase-3 are observed by Western blot analysis. Results The results show that Yantiao Fang could improve cell death induced by LPS and significantly inhibit the apoptosis induced by LPS. The activity of caspase-8 and caspase-3 are activated and the levels of Fas and FasL are improved significantly by LPS. Cell survival of ATII is inhibited by LPS. Yantiao Fang decrease the apoptosis of AT II cells induced by LPS. It inhibited the activity of caspase-8 and caspase-3 and reduced the level of Fas and FasL. Conclusion The mechanisms of Yantiao Fang decreased ATII apoptosis lunduced by LPS may be related to Fas/FasL pathway.
作者
施荣
李甫
王倩
熊旭东
SHI Rong;LI Fu;WANG Qian;XIONG Xu-dong(Emergency Department,Shuguang Hospital Affiliated with University of TCM,Shanghai 201203,China)
出处
《现代中药研究与实践》
CAS
2018年第5期14-18,共5页
Research and Practice on Chinese Medicines
基金
国家自然科学基金(81774057)
北京协和医学基金会睿E(睿意)急诊医学研究专项基金资助项目(R2017008)
上海市级医院新兴前沿技术联合攻关项目(SHDC12017X23)
上海市科学技术委员会科研计划项目(15ZR1442000)
