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Journals / Laboratuvar Hayvanları Bilimi ve Uygulamaları Dergisi / 2026 / Cilt: 6 Sayı: 2

Fecal Extracellular Vesicles from Laboratory Mice Reprogram Neuroinflammatory Responses in Microglia

Laboratuvar farelerinden elde edilen fekal ekstraselüler veziküller, mikroglialarda nöroinflamatuvar yanıtları yeniden programlar

Sinan Gönüllü(BURSA ULUDAG UNIVERSITY)0000-0002-6252-3216
Hande Pelin Özdemir(ATATÜRK ÜNİVERSİTESİ)0009-0004-5187-0687
Sefa Küçükler(ATATURK UNIVERSITY)0000-0002-8222-5515
Selçuk Özdemir(Atatürk University,Erzurum,Turkey)0000-0001-7539-0523
Pages
174–192
Publication Date
2026-09-23
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AI Summary & Key Takeaways

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⚡ TL;DR(Too Long; Didn't Read)

The findings indicated that EV treatment markedly inhibited the production of pro-inflammatory mediators, including interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2).

📌 3 Key Takeaways

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Objective & Hypothesis:This study aims to examine the impact of fecal extracellular vesicles (EVs) derived from laboratory mice on lipopolysaccharide (LPS)-induced neuroinflammatory responses in BV2 microglia cells.
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Key Findings & Data:TEM analysis demonstrated characteristic vesicular morphology, NTA confirmed particle distribution within the nanometer size range, and Western blot analysis indicated flotillin positivity while calnexin was absent as a negative control, thereby validating EV purity. 3-(4,5-dimetiltiyazol-2-il)-2,5-difeniltetrazolyum bromür (MTT) assay for dose optimization in BV2 microglia cells revealed that a concentration of 10 µg/mL was cytotoxic; therefore, 2.5 µg/mL and 5 µg/mL were selected as safe doses for further investigations.
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Conclusion & Impact:The findings indicated that EV treatment markedly inhibited the production of pro-inflammatory mediators, including interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2).
🔬Design:Modeling & Simulation
📍Scope:Comprehensive Sample
⚙️Variables:Fekal EV · nöroinflamasyon · BV2 mikroglia

Abstract

This study aims to examine the impact of fecal extracellular vesicles (EVs) derived from laboratory mice on lipopolysaccharide (LPS)-induced neuroinflammatory responses in BV2 microglia cells. EVs were isolated using size exclusion chromatography (SEC) and characterized through transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA), and Western blot techniques. TEM analysis demonstrated characteristic vesicular morphology, NTA confirmed particle distribution within the nanometer size range, and Western blot analysis indicated flotillin positivity while calnexin was absent as a negative control, thereby validating EV purity. 3-(4,5-dimetiltiyazol-2-il)-2,5-difeniltetrazolyum bromür (MTT) assay for dose optimization in BV2 microglia cells revealed that a concentration of 10 µg/mL was cytotoxic; therefore, 2.5 µg/mL and 5 µg/mL were selected as safe doses for further investigations. The effects of EV administration on LPS-induced inflammatory responses were assessed using RT-qPCR and ELISA. In addition, the expression levels of neuroinflammation- and microglial activation-related markers, including neurofilament light chain (NfL), glial fibrillary acidic protein (GFAP), ionized calcium-binding adapter molecule 1 (Iba1), and triggering receptor expressed on myeloid cells 2 (TREM2), were also analyzed. The findings indicated that EV treatment markedly inhibited the production of pro-inflammatory mediators, including interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2). In addition, fecal EV treatment attenuated the LPS-induced elevation of the anti-inflammatory mediators interleukin-10 (IL-10) and transforming growth factor-β1 (TGF-β1). Furthermore, changes in the expression of neurofilament light chain (NfL), glial fibrillary acidic protein (GFAP), ionized calcium-binding adapter molecule 1 (Iba1), and triggering receptor expressed on myeloid cells 2 (TREM2) supported the modulation of microglial activation and neuroinflammatory processes by EVs. In conclusion, fecal EVs may modulate neuroinflammatory responses by influencing microglial inflammation and could serve as functional communication mediators within the microbiota-gut-brain axis.

Özet

Bu çalışma, laboratuvar farelerinden elde edilen fekal ekstraselüler veziküllerin (EV’ler), BV2 mikroglia hücrelerinde LPS ile indüklenen nöroinflamatuvar yanıtlar üzerindeki etkisini incelemeyi amaçlamaktadır. EV’ler boyut dışlama kromatografisi (SEC) yöntemi ile izole edilmiş ve transmisyon elektron mikroskopisi (TEM), nanopartikül izleme analizi (NTA) ve Western blot teknikleri ile karakterize edilmiştir. TEM analizi karakteristik veziküler morfolojiyi göstermiş, NTA partiküllerin nanometre aralığında dağılımını doğrulamış ve Western blot analizinde flotillin pozitifliği ile birlikte negatif kontrol olarak kullanılan kalneksin’in (calnexin) yokluğu EV saflığını doğrulamıştır. BV2 mikroglia hücrelerinde MTT testi ile yapılan doz optimizasyonu sonucunda 10 µg/mL konsantrasyonun sitotoksik olduğu belirlenmiş; bu nedenle 2.5 µg/mL ve 5 µg/mL güvenli dozlar olarak ileri analizler için seçilmiştir. EV uygulamasının LPS ile indüklenen inflamatuvar yanıtlar üzerindeki etkileri RT-qPCR ve ELISA yöntemleri kullanılarak değerlendirilmiştir. Ayrıca NfL, GFAP, Iba1 ve TREM2 gibi nöroinflamasyon ve mikroglial aktivasyon ile ilişkili belirteçlerin ekspresyon düzeyleri de analiz edilmiştir. Bulgular, EV tedavisinin Il1b, Il6, Tnfa, Nos2 ve Ptgs2 gibi pro-inflamatuvar gen ve proteinlerin üretimini belirgin şekilde inhibe ettiğini, buna karşılık IL-10 ve TGF-β1 gibi anti-inflamatuvar belirteçleri artırdığını göstermiştir. Ayrıca NfL, GAP-43, Iba1 ve TREM2 ekspresyonundaki değişimler, EV’lerin mikroglial aktivasyon ve nöroinflamatuvar süreçleri modüle ettiğini desteklemiştir. Sonuç olarak fekal ekstraselüler veziküllerin, mikroglial inflamasyonu etkileyerek nöroinflamatuvar yanıtları modüle edebileceği ve mikrobiyota–bağırsak–beyin ekseninde fonksiyonel iletişim aracları olarak rol oynayabileceği düşünülmektedir.

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Cite This Article

Gönüllü, S., Özdemir, H. P., Küçükler, S., Özdemir, S. (2026). Fecal Extracellular Vesicles from Laboratory Mice Reprogram Neuroinflammatory Responses in Microglia. Laboratuvar Hayvanları Bilimi ve Uygulamaları Dergisi, Cilt: 6 Sayı: 2, 174–192. https://doi.org/10.62425/jlasp.1978421

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