Abstract
Neferine is the major alkaloid compound isolated from the seed embryos of lotus. Neferine has many pharmacological effects, such as anti-inflammatory, antioxidative stress, and antiapoptotic effects, and it maintains autophagic balance. The purpose of this study was to explore the mechanism by which neferine attenuates autophagy after permanent cerebral ischemia in rats. We performed permanent cerebral ischemia in rats by middle cerebral artery occlusion (pMCAO) for 12 h with or without administration of neferine or nimodipine, a calcium (Ca2+) channel blocker. Neuroprotective effects were determined by evaluating the infarct volume and neurological deficits. Autophagy and its signaling pathway were determined by evaluating the expression of phosphorylated AMP-activated protein kinase alpha (AMPKα), phosphorylated mammalian target of rapamycin (mTOR), beclin-1, microtubule-associated protein 1A/1B-light chain 3 class II (LC3-II), and p62 by western blotting. Autophagosomes were evaluated by transmission electron microscopy. Neferine treatment significantly reduced infarct volumes and improved neurological deficits. Neferine significantly attenuated the upregulation of autophagy-associated proteins such as LC3-II, beclin-1, and p62 as well as autophagosome formation, all of which were induced by pMCAO. Neferine exerted remarkable protection against cerebral ischemia, possibly via the regulation of autophagy mediated by the Ca2+-dependent AMPK/mTOR pathway.
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Acknowledgements
We would like to thank the Nara Institute of Science and Technology, Japan.
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This research was supported by the Faculty of Medicine Research Fund (grant no. 062–2563), Center for Research and Development of Natural Products for Health, Chiang Mai University. We also gratefully acknowledge support from The Thailand Research Fund (DBG6180030) and the Center of Excellence for Innovation in Chemistry, Ministry of Higher Education, Science, Research and Innovation. Jirakhamon Sengking acknowledges the teaching assistant/research assistant financial support from the Graduate School, Chiang Mai University; the Faculty of Medicine Graduate Scholarship, Chiang Mai University; a 2018 scholarship for graduate students from The King Prajadhipok and Queen Rambhai Barni Memorial Foundation; and a 2020 graduate scholarship from the National Research Council of Thailand (NRCT).
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CT conceived the project; JS, PW, NY, and WC performed the experiments; JS, PW, and CT collected data; JS, JT, CT, and CO analyzed the data; CT and AS supervised the project; JS, CO, JT, AS, and CT wrote the manuscript. All authors read and approved the final manuscript.
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Sengking, J., Oka, C., Wicha, P. et al. Neferine Protects Against Brain Damage in Permanent Cerebral Ischemic Rat Associated with Autophagy Suppression and AMPK/mTOR Regulation. Mol Neurobiol 58, 6304–6315 (2021). https://doi.org/10.1007/s12035-021-02554-z
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DOI: https://doi.org/10.1007/s12035-021-02554-z