Department of Physiology, Faculty of Medicine, Hasanuddin University, Makassar 90245, Indonesia.
Clinical Pharmacy Laboratory, Faculty of Pharmacy, Hasanuddin University, Makassar 90245, Indonesia.
Medicina (Kaunas). 2024 Aug 9;60(8):1284. doi: 10.3390/medicina60081284.
The main problem of vascular preservation is the maintenance of vessel graft quality and function following extended storage. Conventional preservation solutions such as histidine-tryptophan-ketoglutarate (HTK) solution, Phosphate-Buffer Solution (PBS), or sodium chloride 0.9% has been shown to be inadequate in preserving vascular physiological function after 3 days of cold storage. This study aimed to evaluate whether adenosine and lidocaine (AL) in a modified Krebs-Henseleit (KH) solution can preserve the function and histological structure of rat aortic rings after 6 days. Thirty-five aortic rings from male Wistar rats (200-300 g) were harvested and immediately immersed in one of the assigned cold preservation solutions: standard KH, modified KH (mod KH) with lower calcium (Ca) and higher magnesium content (Mg) with or without adenosine and lidocaine (mod KH-AL), and modified KH with AL, insulin, and melatonin (Mod KH-ALMI). The contraction and relaxation function of the aortic rings were examined using an isometric force transducer after 6 days of cold preservation. Hematoxylin and eosin staining were used to analyze the rings' histological structure. Vascular contraction and relaxation functions were severely affected after a 6-day cold storage period in standard KH. Modifying the KH solution by reducing the Ca and increasing the Mg levels greatly recovered the vessel functions. The addition of AL or ALMI to the modified KH did not further recover vascular contractility. However, only the addition of AL to the modified KH increased the ACh-induced relaxation at 6 days when compared to the conventional KH, suggesting that endothelium preservation is improved. From histological analysis, it was found that the addition of AL but not ALMI further improved the endothelial lining and the structure of the elastic membrane layers of the preserved vessels after 6 days of cold preservation. The addition of AL to low calcium-high magnesium KH solution significantly enhanced endothelial preservation and improved endothelial-induced relaxation of preserved vessels after 6 days of cold storage.
血管保存的主要问题是在延长储存后保持血管移植物的质量和功能。已证明,传统的保存溶液,如组氨酸-色氨酸-酮戊二酸(HTK)溶液、磷酸盐缓冲液(PBS)或 0.9%氯化钠在冷储存 3 天后不足以保持血管的生理功能。本研究旨在评估腺嘌呤核苷和利多卡因(AL)在改良 Krebs-Henseleit(KH)溶液中是否能在 6 天后保存大鼠主动脉环的功能和组织结构。从雄性 Wistar 大鼠(200-300g)中采集 35 个主动脉环,并立即浸入一种指定的冷保存溶液中:标准 KH、钙(Ca)含量较低和镁(Mg)含量较高的改良 KH(mod KH),以及含或不含腺嘌呤核苷和利多卡因的改良 KH(mod KH-AL),和含 AL、胰岛素和褪黑素的改良 KH(Mod KH-ALMI)。在冷保存 6 天后,使用等长力换能器检查主动脉环的收缩和舒张功能。使用苏木精和伊红染色分析环的组织结构。在标准 KH 中进行 6 天的冷储存后,血管收缩和舒张功能受到严重影响。通过降低 Ca 并增加 Mg 水平来改良 KH 溶液极大地恢复了血管功能。向改良 KH 中添加 AL 或 ALMI 并不能进一步恢复血管收缩性。然而,与常规 KH 相比,仅在改良 KH 中添加 AL 就可以增加 6 天时 ACh 诱导的舒张作用,这表明内皮细胞的保存得到了改善。从组织学分析发现,仅添加 AL 进一步改善了保存血管的内皮衬里和弹性膜层的结构,而添加 ALMI 则没有。在低钙高镁 KH 溶液中添加 AL 可显著增强内皮保存,并改善冷储存 6 天后保存血管的内皮诱导舒张作用。