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Antisecretory factor (AF16) is a potential therapeutic agent for TBI treatment due to its ability to inhibit fluid secretion and decrease inflammation, intracranial pressure, and interstitial fluid build-up, key hallmarks presented in TBI. Here, we investigated the effect of AF16 in an in vitro model of neuronal injury, as well as its impact on key components of the autophagy pathway and mitochondrial dynamics. N2A<jats:sup>wt<\/jats:sup> cells were treated with AF16, injured using a scratch assay, and analysed using confocal microscopy, correlative light and electron microscopy (CLEM), flow cytometry, and western blotting. Our results reveal that AF16 enhances autophagy activity, regulates mitochondrial dynamics, and provides protection as early as 6\u00a0h post-injury. Fluorescently labelled AF16 was observed to localise to lysosomes and the autophagy compartment, suggesting a role for autophagy and mitochondrial quality control in conferring AF16-associated neuronal protection. This study concludes that AF16 has potential as a therapeutic agent for TBI treatment through is regulation of autophagy and mitochondrial dynamics.<\/jats:p>","DOI":"10.1007\/s12031-024-02268-6","type":"journal-article","created":{"date-parts":[[2024,11,7]],"date-time":"2024-11-07T00:03:01Z","timestamp":1730937781000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Antisecretory Factor 16 (AF16): A Promising Avenue for the Treatment of Traumatic Brain Injury\u2014An In Vitro Model Approach"],"prefix":"10.1007","volume":"74","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6856-6014","authenticated-orcid":false,"given":"Nicola","family":"Vahrmeijer","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7558-7916","authenticated-orcid":false,"given":"Jurgen","family":"Kriel","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-9242-7119","authenticated-orcid":false,"given":"Bradley M.","family":"Harrington","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0596-8849","authenticated-orcid":false,"given":"Anton Du Preez","family":"van Staden","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3202-2867","authenticated-orcid":false,"given":"Adriaan Johannes","family":"Vlok","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4518-7011","authenticated-orcid":false,"given":"Lize","family":"Engelbrecht","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3936-6013","authenticated-orcid":false,"given":"Andre","family":"Du Toit","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5517-373X","authenticated-orcid":false,"given":"Ben","family":"Loos","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2024,11,7]]},"reference":[{"issue":"1","key":"2268_CR1","doi-asserted-by":"publisher","first-page":"129","DOI":"10.1007\/s00701-014-2221-7","volume":"157","author":"M Al-Olama","year":"2015","unstructured":"Al-Olama M, Lange S, L\u00f6nnroth I, Gatzinsky K, Jennische E (2015) Uptake of the antisecretory factor peptide AF-16 in rat blood and cerebrospinal fluid and effects on elevated intracranial pressure. 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