{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,13]],"date-time":"2026-03-13T08:37:25Z","timestamp":1773391045550,"version":"3.50.1"},"reference-count":40,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2022,2,16]],"date-time":"2022-02-16T00:00:00Z","timestamp":1644969600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004663","name":"Ministry of Science and Technology","doi-asserted-by":"publisher","award":["MOST 110-2320-B030-005"],"award-info":[{"award-number":["MOST 110-2320-B030-005"]}],"id":[{"id":"10.13039\/501100004663","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Fu Jen Catholic University Hospital","award":["109-FJUH-09"],"award-info":[{"award-number":["109-FJUH-09"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>Excess synaptic glutamate release has pathological consequences, and the inhibition of glutamate release is crucial for neuroprotection. Kaempferol 3-rhamnoside (KR) is a flavonoid isolated from Schima superba with neuroprotective properties, and its effecton the release of glutamate from rat cerebrocortical nerve terminals was investigated. KR produced a concentration-dependent inhibition of 4-aminopyridine (4-AP)-evoked glutamate release with half-maximal inhibitory concentration value of 17 \u00b5M. The inhibition of glutamate release by KR was completely abolished by the omission of external Ca2+ or the depletion of glutamate in synaptic vesicles, and it was unaffected by blocking carrier-mediated release. In addition, KR reduced the 4-AP-evoked increase in Ca2+ concentration, while it did not affect 4-AP-evoked membrane potential depolarization. The application of selective antagonists of voltage-dependent Ca2+ channels revealed that the KR-mediated inhibition of glutamate release involved the suppression of P\/Q-type Ca2+ channel activity. Furthermore, the inhibition of release was abolished by the calmodulin antagonist, W7, and Ca2+\/calmodulin-dependent protein kinase II (CaMKII) inhibitor, KN62, but not by the protein kinase A (PKA) inhibitor, H89, or the protein kinase C (PKC) inhibitor, GF109203X. We also found that KR reduced the 4-AP-induced increase in phosphorylation of CaMKII and its substrate synapsin I. Thus, the effect of KR on evoked glutamate release is likely linked to a decrease in P\/Q-type Ca2+ channel activity, as well as to the consequent reduction in the CaMKII\/synapsin I pathway.<\/jats:p>","DOI":"10.3390\/molecules27041342","type":"journal-article","created":{"date-parts":[[2022,2,16]],"date-time":"2022-02-16T21:36:24Z","timestamp":1645047384000},"page":"1342","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Kaempferol 3-Rhamnoside on Glutamate Release from Rat Cerebrocortical Nerve Terminals Involves P\/Q-Type Ca2+ Channel and Ca2+\/Calmodulin-Dependent Protein Kinase II-Dependent Pathway Suppression"],"prefix":"10.3390","volume":"27","author":[{"given":"Tzu-Kang","family":"Lin","sequence":"first","affiliation":[{"name":"Department of Neurosurgery, Fu Jen Catholic University Hospital, Fu Jen Catholic University, New Taipei City 24205, Taiwan"},{"name":"School of Medicine, Fu Jen Catholic University, New Taipei City 24205, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3478-5451","authenticated-orcid":false,"given":"Chi-Feng","family":"Hung","sequence":"additional","affiliation":[{"name":"School of Medicine, Fu Jen Catholic University, New Taipei City 24205, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4934-9126","authenticated-orcid":false,"given":"Jing-Ru","family":"Weng","sequence":"additional","affiliation":[{"name":"Department of Marine Biotechnology and Resources, National Sun Yat-Sen University, Kaohsiung 80424, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ting-Yang","family":"Hsieh","sequence":"additional","affiliation":[{"name":"P.H.D. Program in Nutrition & Food Science, Fu Jen Catholic University, New Taipei City 24205, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4675-0259","authenticated-orcid":false,"given":"Su-Jane","family":"Wang","sequence":"additional","affiliation":[{"name":"School of Medicine, Fu Jen Catholic University, New Taipei City 24205, Taiwan"},{"name":"Research Center for Chinese Herbal Medicine, College of Human Ecology, Chang Gung University of Science and Technology, Taoyuan 33303, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,2,16]]},"reference":[{"key":"ref_1","first-page":"S7","article-title":"Anatomy and physiology of glutamate in the CNS","volume":"44","author":"Greenamyre","year":"1994","journal-title":"Neurology"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"205","DOI":"10.1016\/0165-6147(90)90116-P","article-title":"Excitatory amino acids and synaptic transmission: The evidence for a physiological function","volume":"11","author":"Headley","year":"1990","journal-title":"Trends Pharmacol. Sci."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"799","DOI":"10.1007\/s00702-014-1180-8","article-title":"Glutamate as a neurotransmitter in the healthy brain","volume":"121","author":"Zhou","year":"2014","journal-title":"J. Neural Transm."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"379","DOI":"10.1016\/0165-6147(90)90184-A","article-title":"Excitatory amino acid neurotoxicity and neurodegenerative disease","volume":"11","author":"Meldrum","year":"1990","journal-title":"Trends Pharmacol. Sci."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"379","DOI":"10.1038\/aps.2009.24","article-title":"Molecular mechanisms of excitotoxicity and their relevance to pathogenesis of neurodegenerative diseases","volume":"30","author":"Dong","year":"2009","journal-title":"Acta Pharmacol. Sin."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"2481","DOI":"10.1111\/j.1460-9568.2007.05873.x","article-title":"Neuroprotectant minocycline depresses glutamatergic neurotransmission and Ca(2+) signalling in hippocampal neurons","volume":"26","author":"Egea","year":"2007","journal-title":"Eur. J. Neurosci."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"38","DOI":"10.1016\/j.neuropharm.2018.09.021","article-title":"Riluzole attenuates the efficacy of glutamatergic transmission by interfering with the size of the readily releasable neurotransmitter pool","volume":"143","author":"Lazarevic","year":"2018","journal-title":"Neuropharmacology"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"77","DOI":"10.1016\/j.neuro.2018.09.004","article-title":"Dexmedetomidine ameliorates lidocaine-induced spinal neurotoxicity via inhibiting glutamate release and the PKC pathway","volume":"69","author":"Xu","year":"2018","journal-title":"Neurotoxicology"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"943","DOI":"10.1142\/S0192415X1650052X","article-title":"Baicalein, a Constituent of Scutellaria baicalensis, Reduces Glutamate Release and Protects Neuronal Cell Against Kainic Acid-Induced Excitotoxicity in Rats","volume":"44","author":"Chang","year":"2016","journal-title":"Am. J. Chin. Med."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"260","DOI":"10.2174\/1570159X17666191101125530","article-title":"Chinese Herbal Medicine Interventions in Neurological Disorder Therapeutics by Regulating Glutamate Signaling","volume":"18","author":"Liu","year":"2020","journal-title":"Curr. Neuropharmacol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"434","DOI":"10.1055\/a-1107-9337","article-title":"11-Keto-\u03b2-Boswellic Acid Attenuates Glutamate Release and Kainic Acid-Induced Excitotoxicity in the Rat Hippocampus","volume":"86","author":"Lu","year":"2020","journal-title":"Planta Med."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Lu, C.W., Lin, T.Y., Pan, T.L., Wang, P.W., Chiu, K.M., Lee, M.Y., and Wang, S.J. (2021). Asiatic Acid Prevents Cognitive Deficits by Inhibiting Calpain Activation and Preserving Synaptic and Mitochondrial Function in Rats with Kainic Acid-Induced Seizure. Biomedicines, 9.","DOI":"10.3390\/biomedicines9030284"},{"key":"ref_13","first-page":"863","article-title":"Chemical constituents in stems of Schima superba","volume":"41","author":"Xu","year":"2010","journal-title":"Chin. Tradit. Herb. Drugs"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"3173","DOI":"10.3390\/molecules19033173","article-title":"Antibacterial effects of afzelin isolated from Cornus macrophylla on Pseudomonas aeruginosa, a leading cause of illness in immunocompromised individuals","volume":"19","author":"Lee","year":"2014","journal-title":"Molecules"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"195","DOI":"10.1111\/bph.13669","article-title":"Afzelin ameliorates D-galactosamine and lipopolysaccharide-induced fulminant hepatic failure by modulating mitochondrial quality control and dynamics","volume":"174","author":"Lee","year":"2017","journal-title":"Br. J. Pharmacol."},{"key":"ref_16","first-page":"2516","article-title":"Afzelin suppresses proinflammatory responses inparticulate matter-exposed human keratinocytes","volume":"43","author":"Kim","year":"2019","journal-title":"Int. J. Mol. Med."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Wu, C., Wu, H.T., Wang, Q., Wang, G.H., Yi, X., Chen, Y.P., and Zhou, G.X. (2019). Anticandidal Potential of Stem Bark Extract from Schima superba and the Identification of Its Major Anticandidal Compound. Molecules, 24.","DOI":"10.3390\/molecules24081587"},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Kim, J.H., Lee, S., and Cho, E.J. (2020). Flavonoids from Acer okamotoanum Inhibit Adipocyte Differentiation and Promote Lipolysis in the 3T3-L1 Cells. Molecules, 25.","DOI":"10.3390\/molecules25081920"},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Radziejewska, I., Supruniuk, K., Czarnomysy, R., Buzun, K., and Bielawska, A. (2021). Anti-Cancer Potential of Afzelin towards AGS Gastric Cancer Cells. Pharmaceuticals, 14.","DOI":"10.3390\/ph14100973"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"9182","DOI":"10.1038\/s41598-021-88463-6","article-title":"Central administration of afzelin extracted from Ribes fasciculatum improves cognitive and memory function in a mouse model of dementia","volume":"11","author":"Oh","year":"2021","journal-title":"Sci. Rep."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"115","DOI":"10.1016\/0006-8993(86)91464-2","article-title":"A rapid method for isolation of synaptosomes on Percoll gradients","volume":"372","author":"Dunkley","year":"1986","journal-title":"Brain Res."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1265","DOI":"10.1023\/A:1007648229795","article-title":"Synaptosomes still viable after 25 years of superfusion","volume":"25","author":"Raiteri","year":"2000","journal-title":"Neurochem. Res."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1016\/S0079-6123(08)60427-6","article-title":"Presynaptic modulation of glutamate release","volume":"116","author":"Nicholls","year":"1998","journal-title":"Prog. Brain Res."},{"key":"ref_24","first-page":"31","article-title":"The role of the plasma membrane and intracellular organelles in synaptosomal calcium regulation","volume":"42","author":"Nicholls","year":"1987","journal-title":"Soc. Gen. Physiol. Ser."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"2009","DOI":"10.1111\/j.1460-9568.1997.tb01369.x","article-title":"Presynaptic modulation of glutamate release targets different calcium channels in rat cerebrocortical nerve terminals","volume":"9","year":"1997","journal-title":"Eur. J. Neurosci."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1016\/S0304-3940(02)00719-X","article-title":"Differential coupling of N- and P\/Q-type calcium channels to glutamate exocytosis in the rat cerebral cortex","volume":"330","year":"2002","journal-title":"Neurosci. Lett."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1016\/S0896-6273(00)80510-3","article-title":"Neuronal calcium signaling","volume":"21","author":"Berridge","year":"1998","journal-title":"Neuron"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"424","DOI":"10.2174\/1389200218666170710190249","article-title":"Natural or Plant Products for the Treatment of Neurological Disorders: Current Knowledge","volume":"19","author":"Parvez","year":"2018","journal-title":"Curr. Drug Metab."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"247","DOI":"10.2174\/1570159X16666180911124605","article-title":"Neuroprotective Strategies for Neurological Disorders by Natural Products: An update","volume":"17","author":"Rehman","year":"2019","journal-title":"Curr. Neuropharmacol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1007\/BF00966922","article-title":"Mechanisms in the regulation of neurotransmitter release from brain nerve terminals: Current hypotheses","volume":"18","author":"Sihra","year":"1993","journal-title":"Neurochem. Res."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1693","DOI":"10.1111\/j.1471-4159.1989.tb09232.x","article-title":"Repetitive action potentials in isolated nerve terminals in the presence of 4-aminopyridine: Effects on cytosolic free Ca2+ and glutamate release","volume":"53","author":"Tibbs","year":"1989","journal-title":"J. Neurochem."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"519","DOI":"10.1016\/S0165-6147(00)01800-9","article-title":"Interactions between presynaptic Ca2+ channels, cytoplasmic messengers and proteins of the synaptic vesicle release complex","volume":"22","author":"Jarvis","year":"2001","journal-title":"Trends Pharmacol. Sci."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"417","DOI":"10.1038\/359417a0","article-title":"Synaptic vesicle-associated Ca2+\/calmodulin-dependent protein kinase II is a binding protein for synapsin I","volume":"359","author":"Benfenati","year":"1992","journal-title":"Nature"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"7944","DOI":"10.1523\/JNEUROSCI.21-20-07944.2001","article-title":"Opposing changes in phosphorylation of specific sites in synapsin I during Ca2+-dependent glutamate release in isolated nerve terminals","volume":"21","author":"Jovanovic","year":"2001","journal-title":"J. Neurosci."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"647","DOI":"10.1038\/343647a0","article-title":"Calcium\/calmodulin-dependent protein kinase II increases glutamate and noradrenaline release from synaptosomes","volume":"343","author":"Nichols","year":"1990","journal-title":"Nature"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"967","DOI":"10.1021\/np50096a027","article-title":"Flavonoids from Koelreuteriahenryi and other sources as protein-tyrosine kinase inhibitors","volume":"56","author":"Ma","year":"1993","journal-title":"J. Nat. Prod."},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Lu, C.W., Lin, T.Y., Chiu, K.M., Lee, M.Y., Huang, J.H., and Wang, S.J. (2020). Silymarin Inhibits Glutamate Release and Prevents against Kainic Acid-Induced Excitotoxic Injury in Rats. Biomedicines, 8.","DOI":"10.3390\/biomedicines8110486"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"3440","DOI":"10.1016\/S0021-9258(19)83641-4","article-title":"A new generation of Ca2+ indicators with greatly improved fluorescence properties","volume":"260","author":"Grynkiewicz","year":"1985","journal-title":"J. Biol. Chem."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1983","DOI":"10.1016\/S0021-9258(18)46043-7","article-title":"Localized Ca2+ entry preferentially effects protein dephosphorylation, phosphorylation, and glutamate release","volume":"267","author":"Sihra","year":"1992","journal-title":"J. Biol. Chem."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"552","DOI":"10.1111\/j.1471-4159.1987.tb04128.x","article-title":"Ionic dependence of membrane potential and glutamate receptor-linked responses in synaptoneurosomes as measured with a cyanine dye, DiS-C2-(5)","volume":"48","author":"Akerman","year":"1987","journal-title":"J. Neurochem."}],"container-title":["Molecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1420-3049\/27\/4\/1342\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T22:21:02Z","timestamp":1760134862000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1420-3049\/27\/4\/1342"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,2,16]]},"references-count":40,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2022,2]]}},"alternative-id":["molecules27041342"],"URL":"https:\/\/doi.org\/10.3390\/molecules27041342","relation":{},"ISSN":["1420-3049"],"issn-type":[{"value":"1420-3049","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,2,16]]}}}