Ulus Travma Acil Cerrahi Derg. 2023; 29(11): 1203-1211 | DOI: 10.14744/tjtes.2023.80743

Histological and electroencephalographic demonstration of probiotic effect for reduce of oxidative stress and apoptosis in experimental traumatic brain injury

Emine Müge Karakayalý¹, Erdoðan Kocamaz², Þüheda Alpay³, Tuna Önal⁴, Mustafa Öztatlýcý⁵, Rabia Duruþma², Hasan Fehmi Özel⁶, Mesut Mete⁷, Mustafa Barutcuoglu⁷, Necip Kutlu³, Mehmet Ýbrahim Tuðlu^2
1Department of Medical Microbiology, Izmir Democracy University Faculty of Medicine, Ýzmir-Türkiye
²Department of Histology and Embryology, Manisa Celal Bayar University Faculty of Medicine, Manisa-Türkiye
³Department of Physiology Department, Manisa Celal Bayar University Faculty of Medicine, Manisa-Türkiye
⁴4Department of Histology and Embryology, Bandýrma Onyedi Eylül,University Faculty of Medicine, Bandýrma-Türkiye
⁵Department of Histology and Embryology, Gaziantep Islam Bilim ve Teknoloji University Faculty of Medicine, Gaziantep-Türkiye
⁶Department of Health Sciences, Celal Bayar University School of Vocation, Manisa-Türkiye
⁷Department of Neurosurgery, Manisa Celal Bayar University Faculty of Medicine, Manisa-Türkiye

BACKGROUND: The gut microbiota modulates nervous system function. In the literature, it has been shown that this modula-tion is used in many nervous system injuries through oxidative stress (OS) and apoptosis mechanisms. In this study, it was aimed to investigate the neuroprotective effects of probiotic (PB) treatment in a rat traumatic brain injury (TBI) model with histological and electroencephalographic (EEG) data.
METHODS: Forty male Wistar albino rats were divided into four groups. Group 1 was the control group (CONTROL, n=10) and no trauma was applied. Group 2 was the trauma group with the weight-drop technique (TBH, n=10). Group 3 was the sham group (SHAM), (TBH+sterile saline [SS], n=10) rats were given 500 µL of SS per day by oral gavage. Group 4 was the PB treatment group, (TBH+PB, n=10) rats were treated daily for 7 days with 500 µL of PB oral gavage. Brain samples were collected 7 days after trauma. Histopathological evaluation of brain samples was done with HE. OS with Endothelial nitric oxide synthase, vascularization with Vas-cular Endothelial Growth Factor, gliosis with S100, and apoptosis with caspase 3 were evaluated immunohistochemically. Apoptotic index was determined with TUNEL. In addition, EEG and somatosensory evoked potential (SEP) recording findings were compared.
RESULTS: It was determined by HE staining that there was a significant (P<0.001) damage in the TBI and sham groups compared to the control group. It was found that PB treatment provided a significant (P<0.01) improvement in the damage created. While OS (P<0.01), gliosis (P<0.01), and apoptosis (P<0.05) decreased with PB treatment, angiogenesis (P<0.01) increased. In support of these findings, in the software-mediated EEG and SUP examination; Delta wave power and theta/alpha ratio increased with TBI and de-creased with PB treatment.
CONCLUSION: The results showed that PB treatment provided a significant improvement in rats by reducing OS, apoptosis, and gliosis and increasing vascularity. To the best of our knowledge in the literature, it was shown for the 1st time that histological results for the treatment of PB were supported by software-mediated EEG and SEP analysis.

Keywords: Electroencephalography, histology; probiotic; traumatic brain injury.

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Deneysel travmatik beyin hasarýnda oluþan oksidatif stres ve apoptozun azalmasýnda probiotik etkisinin histolojik ve elektroensefalografik gösterilmesi

Emine Müge Karakayalý¹, Erdoðan Kocamaz², Þüheda Alpay³, Tuna Önal⁴, Mustafa Öztatlýcý⁵, Rabia Duruþma², Hasan Fehmi Özel⁶, Mesut Mete⁷, Mustafa Barutcuoglu⁷, Necip Kutlu³, Mehmet Ýbrahim Tuðlu^2
1Ýzmir Demokrasi Üniversitesi Týp Fakültesi, Týbbi Mikrobiyoloji Ana Bilim dalý, Ýzmir-Türkiye
²Manisa Celal Bayar Üniversitesi Týp Fakültesi, Histoloji ve Embriyoloji Ana Bilim Dalý, Manisa-Türkiye
³Manisa Celal Bayar Üniversitesi Týp Fakültesi, Fizyoloji Ana Bilim Dalý, Manisa-Türkiye
⁴Bandýrma Onyedi Eylül Üniversitesi Týp Fakültesi, Histoloji ve Embriyoloji Ana Bilim Dalý, Bandýrma-Türkiye
⁵Gaziantep Ýslam Bilim ve Teknoloji Üniversitesi Týp fakültesi,Histoloji ve Embriyoloji Ana Bilim Dalý, Gaziantep-Türkiye
⁶Manisa Celal Bayar Üniversitesi, Saðlýk Meslek Yüksekokulu, Saðlýk Bilimleri,Manisa-Türkiye
⁷Manisa Celal Bayar Üniversitesi Týp Fakültesi, Nöroþirurji Ana Bilim Dalý, Manisa-Türkiye

AMAÇ: Baðýrsak mikrobiyotasý sinir sistemi fonksiyonunu modüle etmektedir. Literatürde bu modülasyonda oksidatif stres ve apoptoz mekanizmalarý üzerinden birçok sinir sistemi hasarýnda kullanýldýðý gösterilmektedir. Bu çalýþmada sýçan travmatik beyin hasarý (TBH) modelinde probiyotik (PB) tedavisine baðlý nöroprotektif etkilerinin histolojik ve elektroensefalografik (EEG) veriler ile araþtýrýlmasý amaçlandý.
GEREÇ VE YÖNTEM: 40 erkek Wistar albino sýçan 4 gruba ayrýldý. Grup 1, Kontrol grubu (KONTROL, n=10) olup travma uygulanmadý. Grup 2, aðýrlýk düþürme tekniði ile yapýlan Travma grubuydu (TBH, n=10). Grup 3, taklit grubu (SHAM) olup (TBH+SS, n=10) sýçanlara günde 500 μl steril salin (SS) oral gavaj ile verildi. Grup 4, PB tedavi grubu olup (TBH+PB, n=10) sýçanlar günlük olarak 500 μl PB oral gavaj ile 7 gün tedavi edildi. Beyin örnekleri travmadan 7 gün sonra toplandý. Beyin örneklerinde histopatolojik deðerlendirme HE ile yapýldý. eNOS ile oksidatif stres, VEGF ile damarlanma, S100 ile gliosis ve kaspaz 3 ile apoptoz ile immünohistokimyasal olarak deðerlendirildi. TUNEL ile apoptotik indeks saptandý. Ek olarak, EEG ve Somatosensoriyal uyarýlmýþ potansiyel (SUP) kayýt bulgularý karþýlaþtýrýldý.
BULGULAR: TBH ve sham gruplarýnda kontrol göre oldukça anlamlý (p<0.001) hasar oluþturulduðu HE boyamalarý ile saptandý. Oluþturulan hasar-da, PB tedavisinin anlamlý (p<0.01) bir iyileþmeyi saðladýðý bulundu. PB tedavisi ile oksidatif stres (p<0.01), gliosis (p<0.01) ve apoptoz (p<0.05) azalýrken, anjiyogenez (p<0.01) arttý. Bu bulgularý destekler biçimde yazýlým aracýlý yapýlan EEG ve SUP incelemesinde; Delta dalga gücü ve teta /alfa oraný TBH ile artarken PB tedavisi ile azaldý.
SONUÇ: Sonuçlar PB tedavisinin sýçanlarda oksidatif stres, apoptoz ve gliosizi azaltýp damarlanmayý artýrarak anlamlý bir iyileþme saðladýðýný gösterdi. Literatürde bildiðimiz kadarý ile ilk defa yazýlým aracýlý EEG ive SUP analizi ile PB tedavisi için histolojik sonuçlarýn desteklendiði gösterildi.

Anahtar Kelimeler: Elektroensefalografi, histoloji; probiyotik; travmatik beyin hasarý.

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