冷冻保存后人乳动脉内皮依赖性特性的保留

Retention of endothelium-dependent properties in human mammary arteries after cryopreservation.

作者信息

Pompilio G, Polvani G L, Antona C, Rossoni G, Guarino A, Porqueddu M, Buche M, Biglioli P, Sala A

机构信息

Department of Cardiac Surgery, Italian Homograft Bank, Milan, Italy.

出版信息

Ann Thorac Surg. 1996 Feb;61(2):667-73. doi: 10.1016/0003-4975(95)01090-4.

DOI:10.1016/0003-4975(95)01090-4

PMID:8572785

Abstract

BACKGROUND

We investigated the effects of cryopreservation and antibiotic treatment on endothelium-dependent vasomotor properties of human internal mammary arteries (IMAs).

METHODS

Sixty IMA specimens from routine coronary artery bypass grafting procedures were randomly assigned to six groups. Group I (controls) were immediately tested after harvest. Remaining groups were prepared according to a stepwise design: group II, 6 hours of warm ischemia; group III, 6 hours of warm ischemia + 24 hours at 4 degrees C (without antibiotics); group IV, 6 hours of warm ischemia + 24 hours of 4 degrees C antibiotic disinfection; group V, 6 hours of warm ischemia + 24 hours at 4 degrees C (without antibiotics) + cryopreservation; and group VI, 6 hours of warm ischemia + 24 hours of 4 degrees C disinfection+cryopreservation. The IMA specimens were cut into rings and the tension of vascular smooth muscle was recorded. The IMA rings were contracted with norepinephrine (3 x 10(-6) mol/L) and tested with cumulative concentrations of acetylcholine (from 1 x 10(-9) to 1 x 10(-5) mol/L), contracted with endothelin-1 (from 1 x 10(-11) to 1 x 10(-6) mol/L), and contracted with the nitric oxide-synthase inhibitor NG-monomethyl-L-arginine (1 x 10(-4) mol/L). Rings were also tested for their capacity to generate 6-keto-prostaglandin F1 (the stable metabolite of prostacyclin), and endothelial cell viability rate was finally evaluated with the trypan blue dye exclusion method.

RESULTS

Our results show that a complete cryopreservation protocol does not significantly modify (p > 0.05) the relaxant activity to acetylcholine in norepinephrine-precontracted IMA rings (controls; 90.2% +/- 4.2% vs group VI, 77.1% +/- 6.2%) or the vasoconstrictor response induced by endothelin-1 (controls, 62.6% +/- 2.8% versus group VI, 73.7% +/- 4.8%) and NG-monomethyl-L-arginine (controls, 22.4% +/- 1.5% versus group VI, 18.9% +/- 1.9%). Furthermore, IMA cryopreservation does not significantly modify (p > 0.05) the endothelial release of prostacyclin either in basal conditions (-20% versus controls) or during pharmacologic intervention with acetylcholine (-18% versus controls), endothelin-1 (-17% versus controls), and NG-monomethyl-L-arginine (-18% versus controls).

CONCLUSIONS

We conclude that the IMA endothelial function does not seem significantly injured by any of the current steps of disinfection and cryopreservation.

摘要

背景

我们研究了冷冻保存和抗生素处理对人乳内动脉（IMA）内皮依赖性血管舒缩特性的影响。

方法

将60个取自常规冠状动脉旁路移植手术的IMA标本随机分为6组。第I组（对照组）在收获后立即进行测试。其余组按照逐步设计进行处理：第II组，6小时热缺血；第III组，6小时热缺血 + 4℃保存24小时（无抗生素）；第IV组，6小时热缺血 + 4℃抗生素消毒24小时；第V组，6小时热缺血 + 4℃保存24小时（无抗生素）+冷冻保存；第VI组，6小时热缺血 + 4℃消毒24小时+冷冻保存。将IMA标本切成环并记录血管平滑肌的张力。用去甲肾上腺素（3×10⁻⁶mol/L）使IMA环收缩，并用累积浓度的乙酰胆碱（从1×10⁻⁹到1×10⁻⁵mol/L）进行测试，用内皮素 - 1（从1×10⁻¹¹到1×10⁻⁶mol/L）使环收缩，并用一氧化氮合酶抑制剂NG - 单甲基 - L - 精氨酸（1×10⁻⁴mol/L）使环收缩。还测试了环产生6 - 酮 - 前列腺素F1（前列环素的稳定代谢产物）的能力，最后用台盼蓝染料排除法评估内皮细胞活力率。

结果

我们的结果表明，完整的冷冻保存方案不会显著改变（p>0.05）去甲肾上腺素预收缩的IMA环对乙酰胆碱的舒张活性（对照组；90.2%±4.2% vs第VI组，77.1%±6.2%）或内皮素 - 1诱导的血管收缩反应（对照组，62.6%±2.8% vs第VI组，73.7%±4.8%）以及NG - 单甲基 - L - 精氨酸诱导的反应（对照组，22.4%±1.5% vs第VI组，18.9%±1.9%）。此外，IMA冷冻保存无论是在基础条件下（与对照组相比降低20%）还是在乙酰胆碱（与对照组相比降低18%）、内皮素 - 1（与对照组相比降低17%）和NG - 单甲基 - L - 精氨酸（与对照组相比降低18%）的药物干预期间，都不会显著改变（p>0.05）前列环素的内皮释放。