The Change Of CD62L And CD34 After Islet Cell Transplantation Via Portal Vein In Diabetes Rats

Objectives:1. To explore the most suitable dosage of STZ with intraperitoneal injection in the rat experiment model of type 1 Diabetes Mellitus.2. To find a method for islet cell separation and purification as well as the evaluation of islet cell quality;To monitor the change of plasma glucose and CD62L(LECAM-1) in peripheral blood before and after pancreas islet cell transplantation (PIT),and the change of CD62L and CD34 in hepatic tissue after PIT;TO strive for a theoretical basis to prevent and cure the rejection .Materials and Methods:1. Building the diabetic rat models: 120 Wistar male rats were divided into five groups equally at random,group A was injected NS intraperitoneally ,group B was injected Phosphate Buffer Saline(PBS) intraperitoneally,group C,group D and group E were injected STZ respectively at a dosage of 55mg/Kg and 65mg/Kg and 75mg/Kg intraperitoneally.To judge the module of diabetic models according to plasma glucose, urine glucose, body weight, water intake, urine and the changes of the general activities after injection.2.①Rats' islet cell separation and purification and the evaluation of quality: Rats' islet cells were digested by the solution of the collagenase V injected through the common bile duct and purified by Ficoll's discontinuous density gradient centrifugation. The identification of purity,motility rate were carried out with DTZ and with AO and PI,combined Glucose -stimulated insulin secretion test.②The change of CD62L and CD34 in peripheral blood or hepatic tissue before and after PIT: 40 Wistar male rats were divided into four groups equally at random, group A without any management; group B was handled only operation; group C,group D were handled operation and injected NS or CsA (3mg/kg/d)+ MMF(20mg/kg/d)respectively intraperitoneally added. After PIT ,to monitor plasma glucose,the expression of CD62L and CD34 in peripheral blood or hepatic tissue.Results:1. The rate of fully diabetic rats injected at a dosage of 65mg/Kg or 75mg/Kg was 91.67%,the former zero death rate and the latter six death in two weeks, otherwise the rate of group A and group B was zero. while the rate of 55mg/Kg group was only 8.33%. After STZ injected , plasma glucose ascensus significantly,body weight no increasing or even declining,water intake and urine volume was significantly higher in group D and E.2.①The yield of islet cells was 430-690 islets/pancreas. The mean yield was 520士70 islets/pancreas. Islet purity was over 85%, islet viability was more than 90%. The insulin release test indicated the islet cells were sensitive to glucose stimulation.②Plasma glucose,the expression of CD62L in peripheral blood or hepatic tissue in group D were significantly higher than those in group C,while CD34 in hepatic tissue in group D was significantly lower than in group C.③Whether immunosuppressants was used or not, there was positive tendency between CD62L in peripheral blood or plasma glucose and CD62L in hepatic tissue and adverse tendency between plasma glucose and CD34 in hepatic tissue.④After PIT, The expression of CD62L was relatively lower and plasma glucose was efficiently controlled in group D, relatively group C. Conclusions:1. STZ is the most suitable drugs established rat insulin-dependent diabetes model of. The rate of fully diabetic rats injected at a dosage of 65mg/Kg is more higher than the 55mg/kg group,more safety and lower infection rat than the 75mg/kg group. The difference in static is significant(p<0.05).2.①Isolation by the collagenase V and purification by Ficoll's discontinuous density gradient centrifugation can produce a high yield of viable purified islet cells in rats.②After PIT, plasma glucose and the expression of CD62L in peripheral blood or plasma glucose in group D were significantly lower than those in group C,which suggested the expression of CD62L may be concerned with the process of rejection.③After PIT, the expression of CD34 in plasma glucose in group D was significantly higher than this in group C,which suggested the process of revascularization after PIT may be concerned with rejection.