Botox Uygulanan Ratlarda Gastrocnemius Kasinin İmmunohistokimyasal ve Stereolojik Olarak İncelenmesi

Yıl 2024, Cilt: 17 Sayı: 3, 166 - 174, 30.12.2024

Mehmet Uğur Delibaş , Gamze Çakmak

https://doi.org/10.52976/vansaglik.1552892

Öz

Yapılan bu çalışma ile gastrocnemius sinistra kasına botox uygulanan erkek ratların 15. ve 21.günlerdeki kasın hacim yoğunluğunun immunohistokimyasal ve stereolojik olarak incelenmesi amaçlandı. Üç aylık 3 değişik grup olmak üzere 30 adet Wistar Albino rat kullanıldı. Kontrol grubu dışındaki diğer iki gruptaki ratların musculus gastrocnemius sinistra’sına 0.1ml serum fizyolojik içerisinde 2.5U Botox A çözeltisi hazırlanarak enjekte edildi. Ratlar % 10’luk tamponlu formaldehitle perfüze edilerek tespit işlemi yapıldı. Ratların musculus gastrocnemius sinistra’ları perfüzyon işleminden bir hafta sonra diseke edildi. Kesitlerin ilk 15’inden rastgele biri seçilerek takiben her 80. kesit sistematik rastgele örnekleme yöntemiyle alındı. Kesitler 5µm kalınlığında ve 8-10 kesit olacak şekilde elde edildi. Hematoksilen eosin ile boyanan kesitler mikroskop altında fotoğraflandı. Cavalieri prensibine uygun şekilde ve Shtereom I paket program kullanılarak musculus gasrocnemius sinistra’ların total hacim değerleri belirlendi. İstatistiksel sonuçlar SPSS (IBM SPSS for Windows, ver.25) paket programı kullanılarak ANOVA testi ile ortaya konuldu. İstatistiki olarak gruplar arası hacim ölçümlerinin anlamlı bir farklılık gösterdiği belirlendi (p<0.05). Ayrıca gastrocnemius kasında Botox A’nın 15. ve 21. günlere ait etkileri immunohistokimyasal olarak Caspase 3 antikor boyası kullanılarak tespit edildi.

Anahtar Kelimeler

Botox, Hacim, İmmunohistokimyal, Musculus gastrocnemius, Stereoloji.

Etik Beyan

This study was carried out by Van Yuzuncu Yil University approved dated 04/29/2021 and numbered 2021/04-21 by van Yuzuncu Yil University Animal experiments Local Ethics Committee.

Destekleyen Kurum

Van Yuzuncu Yil University Scientific Research Projects Coordination Unit

Proje Numarası

TYL-2022-9745

Teşekkür

We thank Van Yuzuncu Yil University Scientific Research Projects Coordination Unit for their support.

Immunohistochemical and Stereological Examination of The Gastrocnemius Muscle In Rats Applied with Botox

Öz

The aim of this study was to examine immunohistochemically and stereologically the volume density of male rats to which botox was applied to the gastrocnemius sinistra muscle, on the 15th and 21st days. 30 Wistar Albino rats, 3 different three-month-old groups, were used. Botox A solution, 2.5U in 0.1ml physiological saline, was prepared and injected into the gastrocnemius sinistra muscle of the rats in the other two groups except the control group. Rats were fixed by perfusion with 10% buffered formaldehyde. Musculus gastrocnemius sinistra of the rats were dissected one week after the perfusion process. Sections were randomly selected among the first 15 sections, and each subsequent 80th section was taken by systematic random sampling method. Sections were obtained with a thickness of 5µm and 8-10 sections. The sections were stained with hematoxylin and eosin, and photographed under a microscope. Total volume values of musculus gasrocnemius sinistra were determined in accordance with the Cavalieri’s Principle and using the Shtereom I package program. It was determined that the volume values between the groups showed a statistically significant difference (p<0.05). In addition, the effects of Botox A on the gastrocnemius muscle on the 15th and 21st days were detected immunohistochemically using Caspase 3 antibody dye.

Anahtar Kelimeler

Botox, Volume, Immunohistochemistry, Musculus gastrocnemius, Stereology.

Proje Numarası

TYL-2022-9745

Kaynakça

• Akkoç F (2016). İnme Sonrası Spastik Hemipleji Hastalarında El Bilek ve Parmak Fleksör Kaslarında Uygulanan Botulinum Toksin Enjeksiyonlarında Ultrason ve Elektramiyografi Rehberliğinin Retrospektif Olarak Karşılaştırılması. Master Thesis, Celal Bayar Üniversitesi Tıp Fakültesi Fiziksel Tıp ve Rehabilitazyon Anabilim Dalı, Manisa.
• Akpınar O, Ozsimsek A, Guzel M, Nazıroglu, M. (2020). Clostridium botulinum neurotoxin A induces apoptosis and mitochondrial oxidative stress via activation of TRPM2 channel signaling pathway in neuroblastoma and glioblastoma tumor cells. Journal of Receptors and Signal Transduction, 40(6), 620-632.
• Baddeley AJ (1991). Stereology In: Spatial Statistics and Digital Image Analysis. Washington DC, Natural Resources Company, 181-216.
• Berne RM, Levy MN, Koeppen BM, Stanton BA (2009). Berne & Levy Physiology. 6th Edition, USA, Elsevier Mosby. Bulam H (2012). The Inhibitory Effect of Platelet Rich Plasma Mesotheraphy on Botulinum Toxin Type-A Induced Skeletal Muscle Paralysis. Master Thesis, Gazi Üniversitesi, Institute of Health Sciences, Ankara.
• Cartee TV, Monheit GD (2011). An overview of botulinum toxins: past, present, and future. Clinics in Plastic Surgery, 38(3), 409-26.
• Cruz-Orive LM (1999). Precision of Cavalieri sections and slices with local errors. Journal of Microscopy, 193(3), 182-198.
• Dressler D, Saberi FA. (2005). Botulinum toxin: Mechanisms of action. European Neurology, 53(1), 3-9.
• Frank J, Erbguth MD (2004). Historical notes on botulism, Clostridium botulinum, botulinum toxin, and the idea of the therapeutic use of the toxin Movement Disorders 19(8), 2-6.
• Ganong WF (2010). Medical Physiology. 23rd Edition, New York, McGraw-Hill, Companies.
• Gundersen HJ, Jensen EB (1987). The efficiency of systematic sampling in stereology and its prediction. Journal of Microscopy, 147(3), 229-263.
• Güleş ME (2018). Botulinum Toksinin Kas Hacmi Üzerine Etkisi; Deneysel Çalışma. Master Thesis, Bezmialem Vakıf Üniversitesi, Institute of Health Science, İstanbul.
• Han Nami, Kim HD, Eom MJ, You JM, Han J, Kim HK, et al. (2013). Proteomic changes in rat gastrocnemius muscle after botulinum toxin an injection. Annals of Rehabilitation Medicine, 37(2), 157-166.
• Hastad DN, Lacy AC (1998). Measurement and Evaluation in Physical Education and Exercise Science. 3rd edition, Boston, Allyn & Bacon.
• Hole JWJ (1981). Human Anatomy and Physiology. 2nd Edition, USA, Wm. C. Brown Company Publishers. Howard CV, Reed MG (2004). Unbiased Stereology: Three-Dimensional Measurement in Microscopy. 2nd Edition, London, Garland Science, 34-39.
• Howard CV (1998). Unbiased Stereology: Three-Dimensional Measurement in Microscopy. 1st Edition, UK, BIOS Scientific Publishers.
• Işık VM (2017). Tavşan anterior auricular kas modelinde botulinum toksin A doz değişken etkisinin; görsel elektronöromiyogrefik ve elektromikroskobik olarak değerlendirilmesi. Master Thesis, Ankara Eğitim ve Araştırma Hastanesi, Plastik, Rekonstrüktif ve Estetik Cerrahi Kliniği, Ankara. Odaci E, Yıldırım S, Bahadır A, Canan S, Sahin B, Bas O et al. (2004). The possible error sources of new stereological methods and their solutions. Turkiye Klinikleri Journal of Medical Sciences, 24, 78-87.
• Sikandar A, Cheema AH, Younus M, Zaneb H (2013). Mycobacterium avium subspecies paratuberculosis multibacillary infection (Johne’s disease) in a Teddy goat. Pakistan Veterinary Journal, 33(2), 260-262.
• Silva EG, Lima JJC, Costa NP (2021). Use of botulinum toxin in hidradenitis suppurativa. Surgical & Cosmetic Dermatology, 13:e20210013.
• Thomas CE, Combs CM (1965). Spinal cord segments. B. Gross structure in the adult monkey. American Journal of Anatomy, 116(1), 205-216.
• Tighe AP, Schiavo G (2013). Botulinum neurotoxins: Mechanism of action. Toxicon, 67, 87-93.
• Van Beek AL, Lim PK, Gear AJL, Pritzkerve MR (2007). Management of vasospastic disorders with botulinum toxin A. Plastic and Reconstructive Surgery, 119(1), 217-226.
• Vander AJ, Sherman JH, Luciano DS (1994). Human Physiology: The Mechanism of Body Function. 6th Edition, New York, McGraw-Hill.
• Weineck J (1998). Sporda İşlevsel Anatomi. (Çev. AS Elmacı) Ankara, Bağırgan Yayınevi