+4 °C’de Krebs-Henseleit Solüsyonunda Soğuk Depolamanın Sıçan Torasik Aortunun Vazoreaktivitesi Üzerindeki Etkileri

Year 2022, Volume: 11 Issue: 3, 456 - 461, 04.10.2022

Sadettin Demirel

https://doi.org/10.53424/balikesirsbd.1141819

Cited By: 1

Abstract

Amaç: Bu çalışma, Krebs-Henseleit solüsyonunda +4 °C’de 3, 6 ve 18 saat süreyle saklanan sıçan torasik aortlarında endotel-bağımlı ve -bağımsız gevşeme yanıtlarının bozulmadan korunup korunamayacağını araştırmayı amaçlamıştır.
Yöntemler: Deneyler izole organ banyosu modeli ve 10-12 haftalık erkek Wistar sıçanlar kullanılarak gerçekleştirilmiştir. Endotel-bağımlı gevşeme faktörlerinin etkisini araştırmak için, endojen prostanoid sentezini inhibe etmek üzere Krebs-Henseleit çözeltisine siklooksijenaz inhibitörü INDO eklenmiştir. Tek doz PE ile submaksimal kasılma yanıtı elde edilmiş ve ardından endotel-bağımlı gevşeme yanıtlarını indüklemek üzere kümülatif olarak (10-9-10-4 M) ACh uygulanmıştır. Ayrıca, PE ile indüklenen prekontraksiyonu takiben SNP kümülatif (10-9-10-5 M) uygulanarak düz kasa bağlı gevşeme yanıtları elde edilmiştir. İstatistiksel anlamlılık düzeyi p<0.05 olarak belirlenmiştir.
Bulgular: Torasik aort segmentleri (0 saat/kontrol), 10-5 M PE’ye karşı 3123 mg’lık ortalama kasılma yanıtı göstermiştir. 3, 6 ve 18 saat saklanan vasküler halkalarda yanıtlar biraz azalmasına rağmen, anlamlı bir fark gözlenmemiştir (p>0.05). Ayrıca, kümülatif uygulanan ACh, endotel-bağımlı gevşeme yanıtlarında anlamlı bir değişikliğe neden olmamıştır (p>0.05). Benzer şekilde, SNP 3, 6 veya 18 saatlik saklamadan sonra aort segmentlerinde endotel-bağımsız gevşeme yanıtlarını modüle etmemiştir (p>0.05).
Sonuç: Bu çalışmada, Krebs-Henseleit solüsyonunda +4 °C’de 3, 6 veya 18 saatlik saklama sürelerinden sonra sıçan torasik aortlarının endotel-bağımlı ve -bağımsız kasılma-gevşeme yanıtlarının bozulmadan korunabildiğine dair ilk fizyolojik bulgular elde edilmiştir.

Keywords

Soğuk Depolama, Sıçan Torasik Aortu, Vasküler Düz Kas, Vazoreaktivite

Effects of Cold Storage in Krebs-Henseleit Solution at +4 °C on Vasoreactivity of the Rat Thoracic Aorta

Abstract

Aim: This study aimed to investigate whether endothelium-dependent and -independent relaxation responses can be preserved intact in the rat thoracic aorta after storage for 3, 6, and 18 hours in Krebs-Henseleit solution at +4 °C.
Methods: Isolated organ bath model and 10-12 weeks old male Wistar rats were used to perform the experiments. To investigate the effect of endothelium-dependent relaxation factors, the cyclooxygenase inhibitor INDO was added to the Krebs-Henseleit solution to inhibit endogenous prostanoid synthesis. Submaximal contraction response was obtained with a single dose of PE and then ACh was administered cumulatively (10-9-10-4 M) to induce endothelium-dependent relaxation responses. Besides, smooth muscle-dependent relaxation responses were obtained by applying SNP cumulatively (10-9-10-5 M) following precontraction induced by PE. The statistical significance level was considered as p<0.05.
Results: Thoracic aorta segments (0 hours, control) exhibited a mean contraction response of 3123 mg against 10-5 M PE. Although the responses slightly reduced in vascular rings stored for 3, 6, and 18 hours, no significant difference was observed (p>0.05). Besides, cumulatively administered ACh did not cause a significant change in endothelium-dependent relaxation responses (p>0.05). Similarly, SNP did not modulate the endothelium-independent relaxation responses in aortic segments after storage for 3, 6, or 18 hours (p>0.05).
Conclusion: In the present study, the first physiological findings have been obtained that the endothelium-dependent and -independent contraction-relaxation responses of rat thoracic aortas can be preserved intact after storage periods of 3, 6, or 18 hours in Krebs-Henseleit solution at +4°C.

Keywords

Cold Storage, Rat Thoracic Aorta, Vascular Smooth Muscle, Vasoreactivity

References

• Amerini, S., Mantelli, L., & Ledda, F. (1995). Enhancement of the vasoconstrictor response to KCL by nitric oxide synthesis inhibition: a comparison with noradrenaline. Pharmacological research, 31(3-4), 175–181. https://doi.org/10.1016/1043-6618(95)80015-8
• Binko, J., Meachem, S., & Majewski, H. (1999). Endothelium removal induces iNOS in rat aorta in organ culture, leading to tissue damage. The American journal of physiology, 276(1), E125–E134. https://doi.org/10.1152/ajpendo.1999.276.1.E125
• Chong, W. C. F., Ong, P. J. L., Hayward, C., Moat, N., & Collins, P. (2001). Effects of storage solutions on in vitro vasoreactivity of radial artery conduits. The Journal of thoracic and cardiovascular surgery, 122(3), 470-475. https://doi.org/10.1067/mtc.2001.115921
• Cupitra, N. I., Calderón, J. C., & Narvaez-Sanchez, R. (2021). Increased receptor expression supports vascular reactivity of the rabbit aorta during preservation. European journal of cardio-thoracic surgery : official journal of the European Association for Cardio-thoracic Surgery, 59(3), 680–687. https://doi.org/10.1093/ejcts/ezaa386
• Dragun, D., Hoff, U., Park, J. K., Qun, Y., Schneider, W., Luft, F. C., & Haller, H. (2001). Prolonged cold preservation augments vascular injury independent of renal transplant immunogenicity and function. Kidney international, 60(3), 1173–1181. https://doi.org/10.1046/j.1523-1755.2001.0600031173.x
• Jahania, M. S., Sanchez, J. A., Narayan, P., Lasley, R. D., & Mentzer, R. M., Jr (1999). Heart preservation for transplantation: principles and strategies. The Annals of thoracic surgery, 68(5), 1983–1987. https://doi.org/10.1016/s0003-4975(99)01028-0
• Jespersen, B., Tykocki, N. R., Watts, S. W., & Cobbett, P. J. (2015). Measurement of smooth muscle function in the isolated tissue bath-applications to pharmacology research. Journal of visualized experiments : JoVE, (95), 52324. https://doi.org/10.3791/52324
• Kristek, F., Török, J., & Šikulová, J. (1993). Morphological and functional alterations in endothelium, smooth muscle, and nerve fibers in rabbit aorta after storage at 4° C. Cryobiology, 30(4), 376-385. https://doi.org/10.1006/cryo.1993.1037
• Ku, D. D., Liu, Q., Norton, P., & Caulfield, J. B. (1994). Cryopreservation of coronary endothelium and endothelial-mediated responses. Cryobiology, 31(1), 82–89. https://doi.org/10.1006/cryo.1994.1010
• Langerak, S. E., Groenink, M., van der Wall, E. E., Wassenaar, C., Vanbavel, E., van Baal, M. C., & Spaan, J. A. (2001). Impact of current cryopreservation procedures on mechanical and functional properties of human aortic homografts. Transplant international : official journal of the European Society for Organ Transplantation, 14(4), 248–255. https://doi.org/10.1007/s001470100309
• Mendizabal, V. E., Feleder, E. C., & Adler-Graschinsky, E. (1999). Effects of the chronic in vivo administration of L-NAME on the contractile responses of the rat perfused mesenteric bed. Journal of autonomic pharmacology, 19(4), 241–248. https://doi.org/10.1046/j.1365-2680.1999.00140.x
• Murphy, C. O., Pan-Chih, Gott, J. P., & Guyton, R. A. (1997). Coronary microvascular reactivity after ischemic cold storage and reperfusion. The Annals of thoracic surgery, 63(1), 20–27. https://doi.org/10.1016/s0003-4975(96)00767-9
• Mühlbacher, F., Langer, F., & Mittermayer, C. (1999). Preservation solutions for transplantation. Transplantation proceedings, 31(5), 2069–2070. https://doi.org/10.1016/s0041-1345(99)00265-1
• Neil, D. A., Lynch, S. V., Hardie, I. R., & Effeney, D. J. (2002). Cold storage preservation and warm ischaemic injury to isolated arterial segments: endothelial cell injury. American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons, 2(5), 400–409. https://doi.org/10.1034/j.1600-6143.2002.20502.x
• Payne, S. J., Benjamin, I. S., & Alexander, B. (2002). Cold storage of rabbit thoracic aorta in University of Wisconsin solution attenuates P2Y(2) purine receptors. Cryobiology, 44(2), 91–102. https://doi.org/10.1016/s0011-2240(02)00010-x
• Piepot, H. A., Pneumatikos, I. A., Groeneveld, A. B., van Lambalgen, A. A., & Sipkema, P. (2002). Cold storage sensitizes rat femoral artery to an endotoxin-induced decrease in endothelium-dependent relaxation. The Journal of surgical research, 105(2), 189–194. https://doi.org/10.1006/jsre.2002.6380
• Rendal Vázquez, M. E., Rodríguez Cabarcos, M., Martínez Santos, M. V., Fernández Mallo, R. O., Sánchez Ibáñez, J., Segura Iglesias, R., Matheu Capó, G., Filgueira Fernández, P., Pértega Díaz, S., Bermúdez González, T., & Andión Núñez, C. (2004). Functional assessment of cryopreserved human femoral arteries for pharmaceutical research. Cell and tissue banking, 5(2), 105–110. https://doi.org/10.1023/B:CATB.0000034085.37937.33
• Rendal Vázquez, M. E., Rodríguez Cabarcos, M., Martínez Santos, M. V., Fernández Mallo, R. O., Sánchez Ibáñez, J., Segura Iglesias, R., Matheu Capó, G., Filgueira Fernández, P., Pértega Díaz, S., Bermúdez González, T., & Andión Núñez, C. (2004). Functional assessment of cryopreserved human aortas for pharmaceutical research. Cell and tissue banking, 5(2), 119–123. https://doi.org/10.1023/B:CATB.0000034087.26468.5a
• Rendal, E., Rodríguez, M., Martínez, M. V., Fernández, R. O., Sánchez, J., Segura, R., Bermúdez, T., Matheu, G., Filgueira, P., Pértega, S., & Andión, C. (2004). Function of cryopreserved pig aortas. The Journal of surgical research, 120(2), 304–311. https://doi.org/10.1016/j.jss.2004.02.004
• Rinaldi G. J. (2001). Influence of several methodological procedures utilized to obtain in vitro vascular preparations on endothelial activity. Endothelium : journal of endothelial cell research, 8(4), 235–242. https://doi.org/10.3109/10623320109090800
• Shibata S. (1969). Effect of prolonged cold storage on the contractile response of strips of rabbit aorta to various agents. Circulation research, 24(2), 179–187. https://doi.org/10.1161/01.res.24.2.179
• Stanke-Labesque, F., Cracowski, J. L., Devillier, P., Caron, F., & Bessard, G. (1999). Functional assessment of rat aorta after cold storage in different media. Fundamental & clinical pharmacology, 13(3), 310–319. https://doi.org/10.1111/j.1472-8206.1999.tb00350.x
• Thyberg J. (1996). Differentiated properties and proliferation of arterial smooth muscle cells in culture. International review of cytology, 169, 183–265. https://doi.org/10.1016/s0074-7696(08)61987-7
• Török, J., Kristek, F., & Mokrásová, M. (1993). Endothelium-dependent relaxation in rabbit aorta after cold storage. European journal of pharmacology, 228(5-6), 313–319. https://doi.org/10.1016/0926-6917(93)90066-y
• Vo, P. A., Reid, J. J., & Rand, M. J. (1992). Attenuation of vasoconstriction by endogenous nitric oxide in rat caudal artery. British journal of pharmacology, 107(4), 1121–1128. https://doi.org/10.1111/j.1476-5381.1992.tb13417.x
• Wise, E. S., Hocking, K. M., Eagle, S., Absi, T., Komalavilas, P., Cheung-Flynn, J., & Brophy, C. M. (2015). Preservation solution impacts physiologic function and cellular viability of human saphenous vein graft. Surgery, 158(2), 537–546. https://doi.org/10.1016/j.surg.2015.03.036
• Yetik-Anacak, G., & Catravas, J. D. (2006). Nitric oxide and the endothelium: history and impact on cardiovascular disease. Vascular pharmacology, 45(5), 268–276. https://doi.org/10.1016/j.vph.2006.08.002