Disertasi

Variasi Intensitas Latihan Fisik pada Tikus Model Stroke Induksi Oklusi Serebri Media Kajian: BDNF, Irisin, Agrin, Sinaptogenesis, dan Taut Saraf Otot = Variation of Intensity in Physical Exercise on Middle Cerebral Artery Occlusion Stroke Model Rats. Studies: BDNF, Irisin, Agrin, Synaptogenesis, and Neuromuscular Junction.

Latar belakang: Stroke merupakan penyebab kematian kedua dan kecacatan tertinggi di dunia. Kemampuan motorik merupakan salah satu masalah utama pasca-stroke. Salah satu pendekatan pemulihan pasca stroke yang berkembang saat ini adalah latihan interval dengan intensitas yang bervariasi. Pada penelitian ini bertujuan untuk mengkaji neuroplastisitas dan marker tingkat perifer pada tikus model stroke iskemik induksi oklusi arteri serebri media (MCAO) intervensi latihan interval dengan intensitas yang bervariasi pada pemulihan fungsi motorik. Metode: Eksperimen in vivo pada tikus Wistar Jantan berat 250-380 gram, umur 22-24 minggu berasal dari Biofarma Bandung. Tikus dibagi 4 kelompok: Sham, MCAO, MCAO+ Moderate-intensity interval training (MIIT) dan MCAO+ High-intensity Interval Training (HIIT). Model stroke iskemik dengan induksi MCAO selama 90 menit. Enam hari sebelum operasi oklusi dan sham diambil darah. Lima hari sampai satu hari semua tikus diberikan pelatihan treadmill. Satu hari sebelum dan satu hari, tujuh hari dan tiga belas hari setelah operasi dilakukan pemerikaan klinis rotarod dan footprint. Satu hari setelah operasi, kelompok MCAO+MIIT dan MCAO+HIIT dilakukan tes kelelahan. Kelompok MCAO+HIIT memperoleh intervensi latihan interval intensitas tinggi selama 2 minggu, sedangkan MCAO+MIIT memperoleh intensitas sedang. Empat belas hari setelah operasi dilakukan necropsy, diambil sampel darah, jaringan otak dan otot gastrocnemius. Sampel darah disentrifugasi untuk memperoleh serum guna pemeriksaan level BDNF, irisin dan agrin dengan ELISA. Jaringan otak dan otot direndam formalin, kemudian dibuat parafin blok. Selanjutnya jaringan otak dilakukan pewarnaan hemaktosilin eosin (HE) dan imunohistokimia Syn-1 dan PSD-95. Jaringan otot dilakukan pewarnanaan HE dan imunoflouresensi TSCs dan AChR. Hasil: HIIT meningkatkan kemampuan motorik tikus pada kelompok MCAO+HIIT. BDNF dan Agrin meningkat pada kelompok MCAO+HIIT dan MCAO+MIIT, dan irisin pada kelompok MCAO+HIIT dan MCAO. Densitas neuron, Syn-1 dan PSD-95 pada kelompok MCAO+HIIT signifikan lebih tinggi dibanding MCAO+MIIT dan MCAO. Terdapat hubungan yang signifikan antara BDNF dengan motorik, BDNF dengan agrin dan Syn-1 dengan PSD-95. Kesimpulan: HIIT pada tikus model oklusi arteri serebri media bemanfaat untuk pemulihan fungsi motorik pasca stroke dengan peningkatan marker neuroplastisitas, sinaptogenesis dan marker perifer. Latihan dengan intensitas sedang juga bermanfaat dengan adanya peningkatan BDNF dan Agrin serta Syn-1. Dengan demikian, latihan interval yang melibatkan intensitas tinggi dan sedang dapat bermanfaat untuk pemulihan pasca-stroke iskemik.
Kata kunci: Agrin, BDNF, Irisin, MCAO, Sinaptogenesis, Taut Saraf Otot


Introduction: Stroke is the second leading cause of death and the highest disability in the world. Impaired motor function is among the primary challenges faced following a stroke. Currently, one of the most advanced methods for post-stroke rehabilitation is high-intensity interval training (HIIT). This study aims to explore neuroplasticity and peripheral markers in a rat model of ischemic stroke induction middle cerebral artery occlusion (MCAO) with highintensity interval training interventions on the restoration of motor function. Methods: In vivo experiments on male Wistar rats weighing 250-380 grams, 22-24 weeks old, sourced from Biofarma Bandung. The rats were divided into 4 groups: Sham, MCAO, MCAO + Moderate-intensity Interval Training (MIIT), and MCAO+HIIT. Ischemic stroke model with induction of MCAO for 90 minutes. Six days before the surgery, blood was drawn. Five days to one day, all the rats were given treadmill training. One day before, one day, seven days, and thirteen days after surgery, a clinical examination of the rotarod and footprint was performed. One day after surgery, the MCAO+MIIT and MCAO+HIIT groups were dose-tested. The MCAO+HIIT group obtained a HIIT for 2 weeks, while the MCAO+MIIT group obtained a MIIT. Fourteen days after the surgery was performed necropsy, samples of blood, brain tissue, and gastrocnemius muscle were taken. Blood samples are centrifuged to obtain serum for checking BDNF, irisin, and agrin levels with ELISA. Brain and muscle tissue are soaked in formalin. Then, paraffin blocks are made. Furthermore, brain tissue was stained with hematoxylin-eosin (HE) and immunohistochemistry Syn-1 and PSD-95. Muscle tissue was stained with HE and immunofluorescence of TSCs and AChR. Results: HIIT improved the motor performance of rats in the MCAO+HIIT group. BDNF and agrin increased in the MCAO+HIIT and MCAO+MIIT groups, and irisin in the MCAO+HIIT and MCAO groups. Neuron density, Syn-1, and PSD-95 in the MCAO+HIIT group were significantly higher than those in MCAO+MIIT and MCAO. There was a significant relationship between BDNF with motoric function, BDNF with agrin and Syn-1 with PSD-95. Conclusion: HIIT in rat models of MCAO is beneficial for recovering post-stroke motor function, with increased markers of neuroplasticity, synaptogenesis, and peripheral markers. MIIT is also beneficial, with an increase in BDNF, agrin, and Syn-1. Thus, interval training involving high and moderate intensity can be beneficial for post-ischemic stroke recovery.
Keywords: Agrin, BDNF, Irisin, Neuromuscular Junction, MCAO, Synaptogenesis