Abstract
Permanent cerebral ischemia is a consequence of prolonged cerebral artery occlusion that results in severe brain damage. Neurotoxicity occurring after ischemia can induce brain tissue damage by destroying cell organelles and their function. Neferine is a natural compound isolated from the seed embryos of the lotus plant and has broad pharmacological effects, including blockading of the calcium channels, anti-oxidative stress, and anti-apoptosis. This study investigated the ability of neferine to reduce brain injury after permanent cerebral occlusion. Permanent cerebral ischemia in rats was induced by instigation of occlusion of the middle cerebral artery for 24 h. The rats were divided into 6 groups: sham, permanent middle cerebral artery occlusion (pMCAO), pMCAO with neferine and nimodipine treatment. To investigate the severity of the injury, the neurological deficit score and morphological alterations were investigated. After 24 h, the rats were evaluated to assess neurological deficit, infarct volume, morphological change, and the number of apoptotic cell deaths. In addition, the brain tissues were examined by western blot analysis to calculate the expression of proteins related to oxidative stress and apoptosis. The data showed that the neurological deficit scores and the infarct volume were significantly reduced in the neferine-treated rats compared to the vehicle group. Treatment with neferine significantly reduced oxidative stress with a measurable decrease in 4-hydroxynonenal (4-HNE), nitric oxide (NO), neuronal nitric oxide (nNOS), and calcium levels and an upregulation of Hsp70 expression. Neferine treatment also significantly decreased apoptosis, with a decrease in Bax and cleaved caspase-3 and an increase in Bcl-2. This study suggested that neferine had a neuroprotective effect on permanent cerebral ischemia in rats by diminishing oxidative stress and apoptosis.
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Funding
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, Chiang Mai, Thailand. 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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Conducting of methodology, data analysis, writing original draft, and revising of the manuscript were by J.S. The data analysis and writing review editing were contributed by J.T., C.O., and A.S. Participation in methodology was by N.Y. and W.C. Conceptualization, validation, formal analysis, writing review editing, and project administration were conducted by C.T. All authors have approved the final manuscript.
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Sengking, J., Oka, C., Yawoot, N. et al. Protective Effect of Neferine in Permanent Cerebral Ischemic Rats via Anti-Oxidative and Anti-Apoptotic Mechanisms. Neurotox Res 40, 1348–1359 (2022). https://doi.org/10.1007/s12640-022-00568-6
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DOI: https://doi.org/10.1007/s12640-022-00568-6