摘要
目的探讨黄芪多糖(APS)对黄嘌呤氧化酶(XOD)诱导的肺癌A549细胞的自噬作用及微管相关蛋白1轻链3B(LC3B)、哺乳动物雷帕霉素靶蛋白(mTOR)、beclin1蛋白表达的影响。方法 A549细胞分为对照组、模型组、100μg/mL APS处理组、200μg/mL APS处理组及400μg/mL APS处理组,除对照组,其余4组均需XOD诱导24 h建立自噬模型。采用透射电镜观察自噬体的形态、单丹磺酰尸胺(MDC)染色荧光显微镜观察自噬体数量的变化, Western blot法检测LC3B、mTOR、beclin1蛋白水平。结果与空白组比较,模型组细胞自噬体数量增多,且LC3B、beclin1蛋白水平升高, mTOR蛋白水平降低;与模型组比较,(200、400)μg/mL APS处理组细胞自噬体数量显著减少, LC3B、beclin1蛋白水平降低, mTOR蛋白水平升高。结论 APS可降低XOD诱导的A549肺癌细胞自噬水平,并降低LC3B、beclin1蛋白表达、增加mTOR蛋白表达。
Objective To investigate the effects of Astragalus polysaccharide(APS) on autophagy and expression of microtubule-associated protein 1 light chain 3B(LC3B), mammalian target of rapamycin(mTOR) and beclin1 in xanthine oxidase(XOD)-induced autophagic model of non-small cell lung cancer A549 cells. Methods A549 cells were divided into five groups: control group, model group, 100, 200 and 400 μg/mL APS-treated group. Except for control group, all groups were administered XOD for 24 hours to establish autophagic models. Morphology of autophagosome was detected by transmission electron microscopy(TEM) and the number was counted by monodansylcadaverine(MDC) staining. The expression levels of LC3B, beclin1 and mTOR were detected by Western blot analysis. Results Compared with the control group, the number of autophagosome in the model group increased;the expression of LC3B and beclin1 significantly increased;while the expression of mTOR significantly decreased. Compared with the model group, the number of autophagosome decreased remarkably;the expression of LC3B and beclin1 severely decreased, and the expression of mTOR obviously increased in 200 or 400 μg/mL APS-treated group. Conclusion APS reduces the level of autophagy, down-regulates the expression of LC3B and beclin1, and increases mTOR expression in the autophagic model of A549 cells induced by XOD.
作者
王雪林
李杨
刘丹
王彦君
明海霞
WANG Xuelin;LI Yang;LIU Dan;WANG Yanjun;MING Haixia(Gansu University of Chinese Medicine,Gansu University of Chinese Medicine,Lanzhou 730000,China;Gansu Provincial Laboratory of Chinese Medicine Pharmacology and Toxicology,Gansu University of Chinese Medicine,Lanzhou 730000,China;Provincial Key Laboratory of Molecular Medicine and Prevention and Treatment of Major Diseases with Traditional Chinese Medicine in Gansu Colleges,Gansu University of Chinese Medicine,Lanzhou 730000,China;Institute of Integrative Traditional Chinese and Western Medicine,Gansu University of Chinese Medicine,Lanzhou 730000,China)
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2019年第7期619-624,共6页
Chinese Journal of Cellular and Molecular Immunology
基金
甘肃省中医药管理局资助项目(GZK-2017-6)
甘肃省高等学校科研项目(2018A-046)
甘肃省高校重大疾病分子医学与中医药防治研究重点实验室开放基金(FZYX17-18-9)
甘肃中医药大学研究生创新基金(CX201833)
