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目的判定在机械分离纯化犬胰岛过程中,胰腺原位灌洗时应用乌司他丁(UTI)的保护作用,观察胰岛的获取产量。方法将20只犬随机分为两组,每组10只。HC-A组:胰腺经腹主动脉原位灌洗时用冷HC-A液2500ml;HC-A+UTI组:胰腺原位灌洗时,HC-A液中加用UTI10000U/kg。两组均经主胰管用4℃胶原酶V控压灌注,在RicordiChamber系统中消化,用Ficoll连续梯度离心纯化。记录消化时间、胰岛外分泌腺包裹率、胰岛纯度、纯化后胰岛在体外用低糖与高糖刺激下胰岛素分泌量、C-肽的释放量及收获的胰岛当量(IEQ),并对纯化后的胰岛进行光镜及电镜观察。结果HC-A+UTI组与HC-A组在胰腺消化时间、胰岛外分泌腺包裹率、胰岛纯度、纯化后的胰岛在体外经低糖与高糖刺激下胰岛素分泌量、C-肽的释放量以及胰岛的形态和结构上比较,差异均无统计学意义(P>0.05)。HC-A组收获胰岛[(3.42±1.47)×104]IEQ,HC-A+UTI组收获胰岛[(6.17±2.86)×104]IEQ,两组差异有统计学意义(P<0.05)。结论在犬胰岛分离和纯化过程中,胰腺获取原位灌洗液中加用UTI能保护胰腺组织,增加胰岛产量。
Objective To determine the protective effect of ulinastatin (UTI) during islet ligation of pancreatic islets in the process of mechanical isolation and purification of canine islets and to observe the yield of islets. Methods Twenty dogs were randomly divided into two groups, 10 in each. Group HC-A: 2500ml cold HC-A solution was used for the in situ lavage of the pancreas through the abdominal aorta; UTI group was added with UTI10000U / kg in the HC-A + UCP group. Both groups were perfused with 4% collagenase V via the main pancreatic duct, digested in RicordiChamber system, and purified by continuous Ficoll gradient centrifugation. The digestion time, the rate of pancreatic exocrine gland encapsulation, the purity of pancreatic islets, insulin secretion, C-peptide release, and islet insulin equivalent (IEQ) were measured in vitro after stimulation of hypoglycemic and hyperglycemic islets. Purified islets Light microscopy and electron microscopy. Results In HC-A + UTI group and HC-A group, the amount of insulin secretion, the release of C-peptide and the amount of insulin secreted by the purified islets in pancreatic digestion time, pancreatic exocrine gland encapsulation efficiency, Islet morphology and structure comparison, the difference was not statistically significant (P> 0.05). HCI was harvested in [(3.42 ± 1.47) × 104] IEQ and HCI + UTI groups [(6.17 ± 2.86) × 104] IEQ, respectively, with significant difference between the two groups (P <0.05). Conclusion During the process of canine islet isolation and purification, pancreatic islets can be protected by adding UTI to the pancreas in situ.