|
Islet transplantation because of its technology is simple, safe, minimally invasive, enables diabetics graduation insulin therapy to maintain normal blood sugar levels, intensive insulin treatment severe hypoglycemia episodes, reduce complications, and therefore become the treatment of type I diabetes ideal method, and is expected to become a fundamental approach to the treatment of diabetes. Islet number and function of islet transplantation is essential. In this study, through the establishment of a pretreatment of the pancreas - pancreatic ductal Protection Act (ie in the pancreas access to the pancreatic duct injection a small amount of an organ preservation solution containing trypsin inhibitor Trypsin Inhibitor University of Wisconsin solution, TIUW liquid) to obtain the number of adequate SD rat islets, islet transplantation diabetes Balb / c nude mice to observe its role in the treatment of diabetes; collagenase digestion the pancreas obtain islet survival for detection of pancreatic duct cells in the pancreas in distribution, the evaluation of the protective effect of pancreatic duct and pancreatic duct Protection Act, their increase islet access to the amount and function to provide an objective basis. The purpose of (1) to explore the ability to improve cold preservation 6h and 24h of rat pancreatic islet pancreatic ductal Protection Act to obtain the amount and function; (2) to evaluate the protective effect of pancreatic ductal pancreatic duct Protection Act. Methods Male Sprague-Dawley rat pancreatic divided into five groups. Group 1: of fresh pancreatic (n = 10); Group 2: Apply the the pancreatic ductal Protection Act only TIUW liquid save 6h pancreas (n = 10); Group 3: Application pancreatic ductal Protection Act and saved with TIUW liquid 6h pancreas group 4: not applied the pancreatic ductal Protection Act only TIUW solution to save the 24h of the pancreas (n = 10); Group 5: Application pancreatic ductal Protection Act and save with TIUW liquid 24h pancreas (n = 10) (n = 10); . (1) The dithizone staining islet morphology and count, trypan blue staining assess islet survival, glucose-stimulated insulin release test evaluation islet isolated biological function. (2) diabetes the nude mice rows SD rats islet transplantation evaluation islet function in vivo biology. (3) HE staining and trypan blue staining assessment of pancreatic duct cell survival, ink staining and immunohistochemical staining paraffin sections observed collagenase distribution. SD rat islets (1) after the separation and purification to obtain the amount of 590 ± 127,272 ± 50,454 ± 65,253 ± 56,447 ± 66 (islet equivalent ± standard deviation); islet mortality, respectively (5.7 ± 4.2)%, (18.3 ± 6.5 )%, (11.7 ± 4.2)%, (26.3 ± 5.6)%, (15.0 ± 5.3)%; stimulation index were 7.32 ± 2.32,4.81 ± 1.17,7.56 ± 2.44,2.88 ± 1.00,5.71 ± 1.90. (2) Diabetes cure rates were 100%, 0%, 100%, 0%, 70%, respectively. (3) pancreatic ductal cell mortality, respectively (5.5 ± 3.0)%, (58.6 ± 9.1)%, (16.8 ± 4.9)%, (64.4 ± 8.1)%, (17.2 ± 3.0)%, Group 3 and Group 5 The results compare the difference was not statistically significant (P gt; 0.05); groups 3 and 5 collagenase distribution than group 2 and group 4 uniform. All results were significant differences (P lt; 0.05 group2 vs. group3 and group4 vs. group5). Conclusions (1) The Law on the Protection of pancreatic ductal cold preservation 6h and 24h rat pancreatic islets access to volume and function. (2) pancreatic ductal Protection Act better preserved the integrity of the pancreatic duct system, and pancreatic ductal cells of the active state does not deteriorate over time; pancreatic ductal system integrity uniformly distributed collagenase premise.
|