Sains Malaysiana 54(10)(2025): 2455-2465

http://doi.org/10.17576/jsm-2025-5410-10

 

a7-Nicotinic Acetylcholine Receptor Activation Mitigates Neuroinflammation Associated with Hypoxia-Reoxygenation Injury in Zebrafish Model

(Pengaktifan Reseptor Asetilkolina a7 Nikotinik Mengurangkan Keradangan Neuro yang Berkaitan dengan Kecederaan Hipoksia-Pengoksigen Semula dalam Model Ikan Zebra)

 

MOHAMMAD YUSUF HASAN^1,†, AZIM HAIKAL MD ROSLAN^1,†, NORAZRINA AZMI¹, NORLINAH MOHAMED IBRAHIM², ALINA ARULSAMY³, VANESSA LIN LIN LEE³, ROSFAIIZAH SIRAN⁴ & MOHD KAISAN MAHADI^1,*

 

¹Centre for Drug Herbal and Development, Faculty of Pharmacy, Universiti Kebangsaan Malaysia, Jalan Raja Muda Abdul Aziz, 50300 Kuala Lumpur, Malaysia
²Department of Medicine, Faculty of Medicine, Universiti Kebangsaan Malaysia, Bandar Tun Razak,
56000 Cheras, Kuala Lumpur, Malaysia
³Neuropharmacology Research Laboratory, Jeffrey Cheah School of Medicine and Health Sciences, Monash University Malaysia, 47500 Bandar Sunway, Selangor, Malaysia
⁴Neuroscience Research Group (NRG), Faculty of Medicine, Jalan Hospital, Universiti Teknologi MARA, Sungai Buloh Campus, 47000 Sungai Buloh, Selangor, Malaysia

 

Received: 6 May 2025/Accepted: 26 July 2025

 

†both authors contributed equally to this work

 

Abstract

Ischemic stroke is a leading cause of death worldwide, where reduced blood flow to brain tissues can cause potential permanent neurological damage. Current treatments, such as intravenous thrombolysis with tissue plasminogen activator and mechanical thrombectomy, aim to restore cerebral blood flow within hours of stroke onset, often associated with ischemic reperfusion injury. Emerging strategies target the α7-nicotinic acetylcholine receptor (α7nAChR) to resolve neuroinflammation in various pathological conditions; however, the therapeutic effects of these strategies in ischemic reperfusion injury remain unknown. This study investigates the neuroprotective and anti-inflammatory effects of α7nAChR activation in zebrafish following ischemia-reperfusion injury. The hypoxia/reoxygenation model was established by perfusing pure nitrogen gas in a hypoxia chamber for 10 min, followed by a 1-h recovery/reoxygenation in the beaker. Gene expression markers for proinflammatory and anti-inflammatory factors were examined using qRT-PCR from the surviving brain tissues. Mitochondrial dehydrogenase activity was measured to investigate the level of brain damage. A six-minute open tank test assessed behaviour, precisely the turning angle, distance travelled, maximum acceleration, and meandering. Hypoxia/reoxygenation significantly increased the expression of proinflammatory markers, such as TNF-α and IL-6, whereas an α7nAChR agonist reduced the expression of these markers. However, there was no discernible improvement in locomotor activity or brain damage in the agonist group, implying that the neurological impairment was not fully reversed following PNU 282987 pre-treatments.

 

Keywords: Inflammation; ischemic reperfusion injury; ischemic stroke; α7nAChR

 

Abstrak

Strok iskemia merupakan salah satu punca utama kematian di seluruh dunia apabila pengurangan aliran darah ke tisu otak boleh menyebabkan kerosakan neurologi kekal. Rawatan semasa seperti trombolisis intravena dengan aktivator plasminogen tisu dan trombektomi mekanikal bertujuan untuk mengembalikan aliran darah serebrum dalam beberapa jam selepas bermulanya strok, yang sering dikaitkan dengan kecederaan reperfusi iskemia. Strategi baharu yang menyasarkan α7-Nikotinik Reseptor Asetilkolina (α7nAChR) untuk menyelesaikan neuroinflamasi dalam pelbagai keadaan patologi sedang diterokai, namun kesan terapeutik dalam kecederaan reperfusi iskemia masih belum diketahui. Penyelidikan ini mengkaji kesan neuropelindung dan anti-radang daripada pengaktifan α7nAChR pada ikan zebra selepas kecederaan iskemia-reperfusi. Model hipoksia/pemulihan oksigen semula diwujudkan dengan menggunakan gas nitrogen tulen yang disalurkan ke dalam ruang hipoksia selama 10 minit, diikuti dengan pemulihan/pemulihan oksigen semula selama 1 jam di dalam bekas. Penanda ekspresi gen bagi faktor pro-radang dan anti-radang dikaji menggunakan qRT-PCR daripada tisu otak yang masih hidup. Aktiviti dehidrogenase mitokondria diukur untuk mengkaji tahap kerosakan otak. Ujian tangki terbuka selama enam minit menilai tingkah laku, khususnya sudut pusingan, jarak perjalanan, pecutan maksimum dan pergerakan berliku. Hipoksia/pemulihan oksigen semula secara signifikan meningkatkan ekspresi penanda pro-radang seperti TNF-α dan IL-6, manakala agonis α7nAChR mengurangkan penanda ini. Namun, tiada peningkatan ketara dalam aktiviti lokomotor dan kerosakan otak pada kumpulan agonis, menunjukkan bahawa kecacatan neurologi tidak dipulihkan sepenuhnya selepas pra-rawatan PNU 282987.

 

Kata kunci: Inflamasi; kecederaan reperfusi iskemia; strok iskemia; α7nAChR

 

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*Corresponding author; email: kaisanmahadi@ukm.edu.my