eng Science and Education Publishing Journal of Food and Nutrition Research 2333-1240 2023-12-01 11 11 700 706 10.12691/jfnr-11-11-6 JFNR202311116 article Liu Zi-shan 1 Zhang Yan-xue 1 Wu Xiao-lin 1 Gu Ning 1 Xu Xin-ying xuxinying2022@126.com, yuzhuq2008@163.com 1 2 3 Yu Zhu-qin xuxinying2022@126.com, yuzhuq2008@163.com 3 4 4 Institute of Integrative Medicine, Qingdao University Medical College, Qingdao 266021, China; Objective: This study aims to delineate whether autophagy plays an critical role in the neuroprotective effects of picroside II in vitro and further explore potential molecular mechanism. Methods: The oxygen glucose deprivation/reoxygenation (OGD/R) cell model were established in SH-SY5Y cells. The cell viability, cells morphologic change and apoptosis were observed by cell counting kit-8 (CCK-8), inverted microscope and flow cytometry respectively. Autophagy-related proteins Beclin 1, microtubule associated protein 1 light chain 3 (LC3), and p62 levels of brain tissues and cells were detected by Western Blot; furthermore, LC3 expression and autophagy flux of cells were further detected by immunofluorescence labeling and GFP-mRFP-LC3 adenovirus transfection. Reactive oxygen species (ROS) levels and phospho-AMPK, phospho-mTOR, phospho-ULK 1 levels were were detected using ROS assay Kit, Western blot and immunocytochemistry, respectively. Results: Picroside II down-regulated of autophagy-related Beclin-1 and LC3 levels, and up-regulated of p62 protein; meanwhile ameliorated the abnormal morphological structures of nerve cells and apoptosis; and increased cell viability. Furthermore, accompanied by decreasing autophagy flux, picroside II prevented the generation of ROS, down-regulated of AMPK and the up-regulated of mTOR and Unc-51-like kinase 1 (ULK1) levels. Conclusion: Picroside II exerts neuroprotective effect against OGD/R injury by inhibiting ROS-mediated AMPK-mTOR-ULK1 autophagy signaling pathway https://pubs.sciepub.com/jfnr/11/11/6/jfnr-11-11-6.pdf autophagy OGD/R AMPK-mTOR-ULK 1 signaling pathway