The Neuroprotective Effect and Possible Mechanism of Picroside II Against Oxygen Glucose Deprivation/Reoxygenation Cell Model

Liu Zi-shan¹, Zhang Yan-xue¹, Wu Xiao-lin¹, Gu Ning¹, Xu Xin-ying^{1, 2,} and Yu Zhu-qin^{1, 3,}

¹Institute of Integrative Medicine, Qingdao University Medical College, Qingdao 266021, China;

²Department of ICU, Jiyang District TCM Hospital, Jinan 251400, China;

³Department of Medicine, The Sixth Peoples’ Hospital of Qingdao, Qingdao 266023, China;

Cite this paper:
Liu Zi-shan, Zhang Yan-xue, Wu Xiao-lin, Gu Ning, Xu Xin-ying and Yu Zhu-qin. The Neuroprotective Effect and Possible Mechanism of Picroside II Against Oxygen Glucose Deprivation/Reoxygenation Cell Model. Journal of Food and Nutrition Research. 2023; 11(11):700-706. doi: 10.12691/jfnr-11-11-6

Abstract

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

Keywords:
autophagy OGD/R AMPK-mTOR-ULK 1 signaling pathway

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References:

[1]	Lee HY, Kim J, Quan W, et al. Autophagy deficiency in myeloid cells increases susceptibility to obesity-induced diabetes and experimental colitis. AUTOPHAGY 2016, 12: 1390-1403.
[2]	Moors TE, Hoozemans JJ, Ingrassia A, et al. Therapeutic potential of autophagy-enhancing agents in Parkinson's disease. MOL NEURODEGENER, 2017, 12: 11-28.
[3]	Wang K, Liu CY, Zhou LY, et al. APF lncRNA regulates autophagy and myocardial infarction by targeting miR-188-3p. NAT COMMUN, 2015, 6: 6779-6789.
[4]	Ding WX, Ni HM, Gao W, et al. Linking of autophagy to ubiquitin-proteasome system is important for the regulation of endoplasmic reticulum stress and cell viability. AM J PATHOL, 2007, 171: 513-524.
[5]	Wang M, Li YJ, Ding Y, et al. Silibinin prevents autophagic cell death upon oxidative stress in cortical neurons and cerebral ischemia-reperfusion injury. MOL NEUROBIOL, 2016, 53: 932-943.
[6]	Shen B, Zhao C, Chen C, et al. Picroside II protects rat lung and A549 cell against LPS-induced inflammation by the NF-kappaB pathway. INFLAMMATION, 2017, 40: 752-761.
[7]	Tiwari SS, Pandey MM, Srivastava S, et al. TLC densitometric quantification of picrosides (picroside-I and picroside-II) in Picrorhiza kurroa and its substitute Picrorhiza scrophulariiflora and their antioxidant studies. BIOMED CHROMATOGR, 2012, 26: 61-68.
[8]	Gao T, Sheng T, Zhang T, et al. Characterization of picroside II metabolites in rats by ultra-high-performance liquid chromatography combined with electrospray ionization quadrupole time-of-flight tandem mass spectrometry. J Pharm Biomed Anal, 2016, 128: 352-359.
[9]	Zhang H, Zhai L, Wang T, Li S, Guo Y. Picroside II Exerts a Neuroprotective Effect by Inhibiting the Mitochondria Cytochrome C Signal Pathway Following Ischemia Reperfusion Injury in Rats. J Mol Neurosci. 2017. 61(2): 267-278.
[10]	Wang T, Zhai L, Zhang H, et al. Picroside II inhibits the MEK-ERK1/2-COX2 signal pathway to prevent cerebral ischemic injury in rats. J MOL NEUROSCI, 2015, 57: 335-351.
[11]	Ma S, Wang X, Lai F, Lou C. The beneficial pharmacological effects and potential mechanisms of picroside II: Evidence of its benefits from in vitro and in vivo. Biomed Pharmacother. 2020. 130: 110421.
[12]	Cao Y, Liu JW, Yu YJ, et al. Synergistic protective effect of picroside II and NGF on PC12 cells against oxidative stress induced by H2O2. PHARMACOL REP, 2007, 59: 573-579.
[13]	Valko M, Leibfritz D, Moncol J, et al. Free radicals and antioxidants in normal physiological functions and human disease. Int J Biochem Cell Biol, 2007, 39: 44-84.
[14]	Hardie DG, Ross FA, Hawley SA. AMPK: a nutrient and energy sensor that maintains energy homeostasis. Nat Rev Mol Cell Biol, 2012, 13: 251-262.
[15]	Yan RY, Wang SJ, Yao GT, et al. The protective effect and its mechanism of 3-n-butylphthalide pretreatment on cerebral ischemia reperfusion injury in rats. Eur Rev Med Pharmacol Sci, 2017, 21: 5275-5282.
[16]	Alers S, Löffler AS, Wesselborg S, et al. Role of AMPK-mTOR-Ulk1/2 in the Regulation of Autophagy: Cross Talk, Shortcuts, and Feedbacks. MOL CELL BIOL, 2012, 32: 2-11.
[17]	Egan D, Kim J, Shaw RJ, et al. The autophagy initiating kinase ULK1 is regulated via opposing phosphorylation by AMPK and mTOR. AUTOPHAGY, 2011, 7: 643-644.
[18]	Dong F, Yao R, Yu H, et al. Neuroprotection of Ro25-6981 against ischemia/ reperfusion-induced brain injury via inhibition of autophagy. CELL MOL NEUROBIOL, 2017, 37: 743-752.
[19]	Chen Y, Fan Z, Wu Q. Dexmedetomidine improves oxygen-glucose deprivation/reoxygenation (OGD/R) -induced neurological injury through regulating SNHG11/miR-324-3p/VEGFA axis. Bioengineered. 2021. 12(1): 4794-4804.
[20]	Huang J, Canadien V, Lam GY, et al. Activation of antibacterial autophagy by NADPH oxidases. Proc Natl Acad Sci U S A, 2009, 106: 6226-6231.
[21]	Klionsky DJ, Abdalla FC, Abeliovich H, et al. Guidelines for the use and interpretation of assays for monitoring autophagy. AUTOPHAGY, 2012, 8: 445-544.
[22]	Hou Z, Zhou Y and Yang H, et al. Alpha7 nicotinic acetylcholine receptor activation protects against myocardial reperfusion injury through modulation of autophagy. Biochem Biophys Res Commun, 2018, 500: 357-364.
[23]	Hua R, Han S, Zhang N, et al. cPKCgamma-modulated sequential reactivation of mTOR inhibited autophagic flux in neurons exposed to oxygen glucose deprivation/reperfusion. INT J MOL SCI, 2018, 19: 1380-1390.
[24]	Wang JF, Mei ZG, Fu Y, et al. Puerarin protects rat brain against ischemia/reperfusion injury by suppressing autophagy via the AMPK-mTOR-ULK1 signaling pathway. NEURAL REGEN RES, 2018, 13: 989-998.
[25]	Sun L, Zhao M, Liu A, et al. Shear stress induces phenotypic modulation of vascular smooth muscle cells via AMPK/mTOR/ULK1-mediated autophagy. CELL MOL NEUROBIOL, 2018, 38: 541-548.
[26]	Voogd E, Frega M, Hofmeijer J. Neuronal Responses to Ischemia: Scoping Review of Insights from Human-Derived In Vitro Models. Cell Mol Neurobiol. 2023. 43(7): 3137-3160.
[27]	Rolf J, Zarrouk M, Finlay DK, et al. AMPKalpha1: a glucose sensor that controls CD8 T-cell memory. EUR J IMMUNOL, 2013, 43: 889-896.
[28]	Kim YC, Guan KL. mTOR: a pharmacologic target for autophagy regulation. J CLIN INVEST, 2015, 125: 25-32.
[29]	Zhang Y, Miao JM. Ginkgolide K promotes astrocyte proliferation and migration after oxygen-glucose deprivation via inducing protective autophagy through the AMPK/mTOR/ULK1 signaling pathway. EUR J PHARMACOL, 2018, 832: 96-103.
[30]	Itakura E, Mizushima N. Characterization of autophagosome formation site by a hierarchical analysis of mammalian Atg proteins. AUTOPHAGY, 2010, 6: 764-776.