Avila J G, Wang Y, Barbaro B, Gangemi A, Qi M, Kuechle J, Doubleday N, Doubleday M, Churchill T, Salehi P, Shapiro J, Philipson L H, Benedetti E, Lakey J R T, Oberholzer J
Division of Transplantation, University of Illinois at Chicago, Illinois, USA.
Am J Transplant. 2006 Dec;6(12):2861-70. doi: 10.1111/j.1600-6143.2006.01551.x.
During isolation, islets are exposed to warm ischemia. In this study, intraductal administration of oxygenated polymerized, stroma-free hemoglobin-pyridoxalated (Poly SFH-P) was performed to improve O2 delivery. Rat pancreata subjected to 30-min warm ischemia were perfused intraductally with collagenase in oxygenated Poly SFH-P/RPMI or RPMI (control). PO2 was increased by Poly SFH-P (381.7 +/- 35.3 mmHg vs. 202.3 +/- 28.2, p = 0.01) and pH maintained within physiological range (7.4-7.2 vs. 7.1-6.6, p = 0.009). Islet viability (77% +/- 4.6 vs. 63% +/- 4.7, p = 0.04) was improved and apoptosis lower with Poly SFH-P (caspase-3: 34,714 +/- 2167 vs. 45,985 +/- 1382, respectively, p = 0.01). Poly SFH-P improved islet responsiveness to glucose as determined by increased intracellular Ca2+ levels and improved insulin secretion (SI 5.4 +/- 0.1 vs. 3.1 +/- 0.2, p = 0.03). Mitochondrial integrity was improved in Poly SFH-P-treated islets, which showed higher percentage change in membrane potential after glucose stimulation (14.7% +/- 1.8 vs. 9.8 +/- 1.4, respectively, p < 0.05). O2 delivery by Poly SFH-P did not increase oxidative stress (GSH 7.1 +/- 2.9 nm/mg protein for Poly SFH-P vs. 6.8 +/- 2.4 control, p = 0.9) or oxidative injury (MDA 1.8 +/- 0.9 nmol/mg protein vs. 6.2 +/- 2.4, p = 0.19). Time to reach normoglycemia in transplanted diabetic nude mice was shorter (1.8 +/- 0.4 vs. 7 +/- 2.5 days, p = 0.02), and glucose tolerance improved in the Poly SFH-P group (AUC 8106 +/- 590 vs. 10,863 +/- 946, p = 0.03). Oxygenated Poly SFH-P improves islet isolation and transplantation outcomes by preserving mitochondrial integrity.
在胰岛分离过程中，胰岛会经历热缺血。在本研究中，通过导管内注入氧合聚合的、无基质的吡哆醛化血红蛋白（Poly SFH-P）来改善氧气输送。对经历30分钟热缺血的大鼠胰腺，经导管分别用含氧量合Poly SFH-P的RPMI溶液或RPMI溶液（对照）灌注胶原酶。Poly SFH-P使PO2升高（381.7±35.3 mmHg对202.3±28.2，p = 0.01），pH维持在生理范围内（7.4 - 7.2对7.1 - 6.6，p = 0.009）。Poly SFH-P提高了胰岛活力（77%±4.6对63%±4.7，p = 0.04），并降低了细胞凋亡（半胱天冬酶-3分别为34,714±2167对45,985±1382，p = 0.01）。通过细胞内Ca2+水平升高和胰岛素分泌改善来确定，Poly SFH-P提高了胰岛对葡萄糖的反应性（刺激指数5.4±0.1对3.1±0.2，p = 0.03）。Poly SFH-P处理的胰岛中线粒体完整性得到改善，在葡萄糖刺激后膜电位变化百分比更高（分别为14.7%±1.8对9.8±1.4，p < 0.05）。Poly SFH-P输送氧气并未增加氧化应激（Poly SFH-P组谷胱甘肽为7.1±2.9 nmol/mg蛋白，对照组为6.8±2.4，p = 0.9）或氧化损伤（丙二醛分别为1.8±0.9 nmol/mg蛋白对6.2±2.4，p = 0.19）。移植后的糖尿病裸鼠达到正常血糖的时间更短（1.8±0.4天对7±2.5天，p = 0.02），且Poly SFH-P组的葡萄糖耐量得到改善（曲线下面积8106±590对10,863±946，p = 0.03）。氧合Poly SFH-P通过保持线粒体完整性改善了胰岛分离和移植结果。
