Effects of isoflurane, ketamine and dantrolene on apoptosis in the rat hippocampus

Year 2020, Volume: 11 Issue: 4, 255 - 261, 30.09.2020

Seyfi Kartal Berrin Günaydın Süreyya Barun Çiğdem Elmas

https://doi.org/10.18663/tjcl.782524

Abstract

Aim: Since both isoflurane and ketamine were already known to cause neurodegenerative apoptotic effects and dantrolene was known to inhibit degeneration, we investigated whether dantrolene could play a cytoprotective role under isoflurane and/or ketamine anesthesia. Therefore, we aimed to determine caspase activation as a marker of apoptosis in hippocampus (CA1 and dentate gyrus regions) of rats exposed to either administration of isoflurane with or without ketamine and dantrolene or isoflurane+ketamine+dantrolene.
Material and Methods: Thirty Wistar male rats were randomly assigned to five groups. Only oxygen 100% was administered into the closed cage for 2 hours in the control group (group A) whereas in the four study groups (as B,C,D and E), either 1.4% isoflurane alone in 100% oxygen was administered (group B) or 1.4% isoflurane in 100% oxygen was administered 60 minutes after intraperitoneal (ip) injection of dantrolene 10 mg/kg (group C), subcutaneous (sc) ketamine 40 mg/kg (group D) or ip dantrolene + sc ketamine (group E). Rats were sacrified to perform histopathologic and immunohistochemical analysis (hematoxylin staining caspase activation).
Results: Isoflurane alone (group B) and isoflurane+ketamine (group D) exposure to rats resulted in a significantly increased caspase activation when compared to control (group A) and dantrolene inhibited isoflurane + ketamine induced apoptosis in the hippocampus. Conclusion: Isoflurane with or without ketamine caused neuroapoptosis in rats and dantrolene attenuated the apoptotic effect of both isoflurane and isoflurane+ketamine by decreasing caspase activation. These results might have an important promising role in anesthetic choice for specific susceptible group after further clinical studies.

Keywords

Isoflurane, ketamine, dantrolene, apoptosis

Supporting Institution

TUBİTAK

Project Number

110S514

Sıçan hipokampüsünde izofluran, ketamin ve dantrolenin apoptozis üzerine etkileri

Abstract

Amaç: Hem izofluran ve ketaminin nörodejeneratif apoptotik etkileri gösterilmiş hem de dantrolenin dejenerasyonu inhibe ettiği bilindiği için biz isoflurane ve/veya ketamine anestezisi altında dantrolenin sitoprotekif bir rolü olup olamayacağını araştırdık. Bu nedenle sıçan hipokampüsünde (CA1 ve dentat girus bölgelerinde) apoptozisin belirteci olan kaspaz aktivasyonunu; izofluranı ketamine ve dantrolen varken veya yokken veya izofluran+ketamin+dantrolen varlığında belirlemeyi amaçladık
Gereç ve Yöntemler: Wistar 30 erkek sıçan rastgele beş gruba ayrıldı. Kontrol grubunda 2 saat sadece %100 oksijen verilirken, dört çalışma grubunda (A.B,C ve D) ya sadece %100 oksijen içinde %1.4 izofluran (grup B) veya intraperitoneal (ip) dantrolen 10 mg/kg enjeksiyonundan (grup C), subkutan (sk) ketamin 40 mg/kg enjeksiyonundan (group D) ya da ip dantrolen + sk ketamine enjeksiyonundan (group E) 60 dk sonra %100 oksijen içinde %1.4 izofluran uygulanmıştır. Sonra sıçanlar histopatolojik ve immunohistokimyasal inceleme (hematoxylin boyama ile kaspas aktivasyonu) için sakrifiye edilmişlerdir.
Bulgular: Sadece izofluran (grup B) ve izofluran+ketamine (grup D) maruz kalan sıçanlarda kontrol grubuna (grup A) göre hipokampüsde kaspas aktivasyonunda önemli artış ile dantrolenin izofluran+ketaminle indüklenmiş apoptozisi inhibe ettiği gözlendi.
Sonuç: İzofluranın ketamine varken ya da yokken sıçanda nöroapoptozise neden olduğu ve dantrolenle; izofluran ve izofluran+ketaminin apoptotik etkisinin azalan kaspaz aktivasyonuyla olduğu gösterilmiştir. Bu sonuçlar ileri klinik çalışmalardan sonra duyarlı grupların anestezi seçimde rol oynabilir.

Keywords

Izofluran, ketamin, dantrolen, apoptozis

Project Number

110S514

References

• 1. Istaphanous GK, Loepke AW. General anesthetics and the developing brain. Curr Opin Anaesthesiol 2009; 22: 368-73.
• 2. Schifilliti D, Grasso G. Conti A, Fodale V. Anaesthetic-related neuroprotection Intravenous or inhalational agents? Drugs 2010; 24: 893-907.
• 3. Stratmann G, Sall JW, May LD, Loepke AW, Lee MT. Beyond anesthetic properties: the effects of isoflurane on brain cell death, neurogenesis, and long-term neurocognitive function. Anesth Analg 2010; 110: 431-7.
• 4. Stratmann G, Sall JW, May LD, et al. Isoflurane differentially affects neurogenesis and long-term neurocognitive function in 60-day old and 7-day old rats. Anesthesiology 2009; 110: 834-48.
• 5. Soriano SG, Liu Q, Li J et al. Ketamine activates cell cycle signaling and apoptosis in the neonatal rat brain. Anesthesiology 2010; 112:1155-63.
• 6. Budihardjo I, Oliver H, Lutter M, Luo X, Wang X. Biochemical pathways of caspase activation during apoptosis. Annu Cell Dev Biol 1999; 15: 269-90.
• 7. Wei H, Kang B, Wei W et al. Isoflurane and sevoflurane affect cell survival and BCL=2/BAX ratio differently. Brain Res 2005; 1037: 139-47.
• 8. Kenneth S, McCarty Jr, Szabo E, Flowers JL, Cox EB, Leight GS. Use of a monoclonal anti-estrogen receptor antibody in the immunohistochemical evaluation of human tumors. Cancer Res 1986; 46:4244-4248
• 9. Xie Z, Culley DJ, Dong Y et al. The common inhalation anesthetic isoflurane induces caspase activation and increases amyloid β-protein level in vivo. Ann Neurol 2008; 64:618-27.
• 10. Valentim AM, Giminiani PD, Ribeiro PO, Rodrigues P, Olsson AS, Antunes LM. Lower isoflurane concentration affects spatial learning and neurodegeneration in adult mice compared with higher concentrations. Anesthesiology 2010; 113:1099-108.
• 11. Zou X, Patterson TA, Sadovova N et al. Potential neurotoxicity of ketamine in the developing rat brain. Toxicol Sci 2009;108: 149-58.
• 12. Mak YT, Lam WP, Lü L, Wong YW, Yew DT. The Toxic Effect of ketamine on SH-SY5Y neuroblastoma cell line and human neuron. Mic Res Tech 2010; 73:195-201.
• 13. Wakasugi M, Hirota K, Roth SH, Ito Y. The effects of general anesthetics on excitatory and inhibitory synaptic transmission in Area CA1 of the rat hippocampus in vitro. Anesth Analg 1999; 88: 676-80.
• 14. Miller KW. The nature of sites of general anesthetic action. Br J Anaesth 2002; 89: 17-31.
• 15. Braun S, Gaza N, Werdenhausen R et al. Ketamine induces apoptosis via the mitochondrial pathway in human lymphocytes and neuronal cells. Br J Anaesth 2010; 105: 347-54.
• 16. Li F, Hayashi T, Jin G et al. The protective effect of dantrolene on ischemic neuronal cell death is associated with reduced expression of endoplasmic reticulum stres markers. Brain Res 2005; 1048:58-65
• 17. Gwak M, Park P, Kim K et al. The effects of dantrolene on hypoxic-ischemic injury in the neonatal rat brain. Neurosurg Anesthesiol 2008; 106:227-33.
• 18. Miura Y1, Grocott HP, Bart RD, Pearlstein RD, Dexter F, Warner DS. Differential effects of anesthetic agents on outcome from near-complete but not incomplete global ischemia in the rat. Anesthesiology. 1998; 89(2):391-400.
• 19. Hare GMT. Xenon anesthesia: safe, protective and neurotoxic? Can J Anesth 2008; 55: 403-7.
• 20. De Tina A, Palanisamy A. General anesthesia during the third trimester: any link to neurocognitive outcomes? Anesthesiol Clin 2017; 35:69-80.