胰导管保护法提高胰岛获取量的实验研究

目的:探讨联合应用胰蛋白酶抑制剂和UW液的胰导管保护方法能否提高保存6 h和24 h大鼠胰腺中胰岛的获取量。方法:从雄性SD大鼠分离得到胰腺,分成5组:组1:新鲜的未用胰导管保护和UW液保存(n=8);组2:末用胰导管保护方法仅用UW液保存6 h(n=8);组3:应用胰导管保护方法并用UW液保存6 h(n=8);组4:未用胰导管保护方法仅用UW液保存24 h(n=8):组5:应用胰导管保护方法并用UW液保存24 h(n=8)。结果:分离纯化后每组的胰岛数量分别为1 820±639,585±151,1 496±676,567±182,1 442±857(胰岛当量±标准差);胰岛存活率分别为(93.8±4.9)%,(82.1±5.6)%,(89.6±4.9)%,(77.9±5.0)%,(86.7±7.1)%;本实验中未应用胰导管保护方法组和应用胰导管保护方法组间差异有显著性(P<0.05,组2和组3相比较,组4和组5相比较)结论:联合应用胰蛋白酶抑制剂和UW液的胰导管保护方法提高了保存6 h和保存24 h胰腺中的胰岛获取量。

Improving islet yield by pancreatic ductal preservation

Objective:To investigate whether pancreatic ductal presservation with trypsin inhibitor and University of Wisconsin solution(UW solution) would improve islet yield from rat pancreases preserved for 6 and 24 h. Methods:Islets were isolated from Sprague-Dawley(SD) rats. Pancreases were classified into five groups:fresh without pancreatic ductal preservation and preserving in University of Wisconsin(group 1,n = 8);preserved for 6 h in UW solution without and with pancreatic ductal preservation(group 2,n = 8 and group 3,n = 8);preserved for 24 h in UW solution without and with pancreatic ductal preservation(group 4,n = 8 and group 5,n = 8). Dithizon(DTZ) staining was used to identify islet yield;trypan blue uptake was used to assess the viability of islets. Results:Islet yields per pancreas in groups 1 to 5 were 1 820 ± 639,585 ± 151,1 496 ± 676,567 ± 182,and 1 442 ± 857(islet equivalent ± SD),respectively. The viability of islets in groups 1 to 5 were(93.8 ± 4.9)%,(82.1 ± 5.6)%,(89.6 ± 4.9)%,(77.9 ± 5.0)% and(86.7 ± 7.1)%,respectively. In all experiments,the differences were significant between the groups with/without pancreatic ductal preservation(P < 0.05,group 2 vs. group 3 and group 4 vs. group 5). Conclusion:Pancreatic ductal preservation with trypsin inhibitor and UW solution improved islet yield after 6 and 24 h of preservation.