Experimental Study On Allogeneic Transplantation Of Mice Islets

MICE ISLET OF RESEARCH ON SEPARATION, PURIFICATION AND DETECTIONObjectives:To purify islet cells.Methods:The islets and pancreatic exocrine cells were collected,and then use three kinds of ways to test the function of islet cells.Results:islets (150)could be obtained from each mouse,and two types of indicators were above 90%,the insulin release amount after the stimulation from glucose increased obviously.Conclusions:Islet isolation and purification methods of the present study is proposed, which can obtain high purity and good function of islets.ESTABLISH OF ISLET CELL TRANSPLANTATION ANIMAL MODEL TOGETHER WITH THE PANCREATIC EXOCRINE CELLS UNDERTHE LEFT RENAL CAPSULE IN DIABETIC MICEObjective:To establish animal model through islets and pancreatic exocrine cells transplantation under the left renal capsule of mice with allogeneic diabetes,and to investigate the damage of pancreatic exocrine cells of isletsMethods:The islets and pancreatic exocrine cells were collected.Simple transplantation group (n=10),250 islets were transplanted into left kidney subcapsule of diabetic mice,common transplantation group (n=10),250 islets and the equal volume of pancreatic exocrine cells were transplanted into different regions of left kidney subcapsule,blood glucose level was monitored.Nephrectomies were performed after 28 days.Result:After islets transplantation, the blood glucose levels in Simple transplantation group and common transplantation group were normal, but a delayed islet function in reversing diabetes was in the common transplantation group,and each mice in both groups became hyperglycemic after nephrectomy.Conclusion:(1)The successful establishment of animal model for islet and pancreatic exocrine cells transplantation under the left renal capsule in diabetic mice.(2)The pancreatic exocrine cells has detrimental effect on islet function after transplantation.PROTECTIVE EFFECTS OF SOMATOSTATIN ON MICE ISLETS INJURY AFTER TRANSPLANTATION BY PANCREAS EXOCRINE CELLSObjective:To investigate protective effects of somatostatin(SS) on mice islets injury after transplantation by pancreas exocrine cells and its mechanismMethods:(1)In vitro,20 male BALB/C mice, the mice were randomly divided into the experimental group (n=10) and the control group (n=10), mice in experimental group was injected with SS(10ug/g) by intraperitoneal injection(i.p) before 30 minutes, the control group was injected with the same amount of normal saline(i.p), the pancreas exocrine cells and islet cells of two groups were extracted respectively,the two kinds of cell apoptosis were detected by flow cytometric. (2)In vivo,8-9 weeks olds male BALB/C mice were induced into diabetic mice with STZ(190mg/kg body weight,i.p)and randomly divided into two groups. In experimental(n=20) and control(n=20) group,250 islets and the equal volume of pancreatic exocrine cells were transplanted into different regions of left kidney subcapsule. The experimental group was injected with SS (10ug/g,tid,i.p) for 28 days after operation,the control group was injected wih the same amount of normal saline(tid,i.p)for 28 days.Then two groups of mice were injected EDU (5ug/g,qd,i.p)for 28 days, glucose tolerance test was performed after 8 days,the left kidney were removed respectively after 10 days and 28 days.The expression of anti-amylase antibodies in subcapsule was detected by using irnmunohistochemieal staining, the proliferation of islet beta cells were detected by using immunofluorescence staining,blood glucose level was monitored continuely.Result:(1) The apoptosis rate of pancreas exocrine cells in the experimental group were significantly higher than the control group(P<0.05),the apoptosis rate of islet cell has little difference between the experimental group and the control group(P>0.05). After islets transplantation,the blood glucose levels in control and experimental group was normal,but a advanced islet function in reversing diabetes was in the experimental group,the average blood glucose level of control group was significantly higher than the experimental group, the blood glucose regulating function of islet is normal.A large number of anti-amylase antibody-positive cells were found in renal subcapsule in the control group while little seen in the experimental group after 10 days, Immunofluorescence showed that the Insulin+/EDU+β of islet in the experimental group were more than the control group.The number of anti-amylase antibody-positive cells in the experimental group were significantly less than the control group after 28 days,but each group had no obvious difference compared with 10 days. The number of increased beta cells in the experimental group were still significantly higher than the control group,but compared with 10 days, the proliferation rate were reduced.Conclusion:SS could induce the apoptosis of damaged pancreas exocrine cells, but the islet cells is less affected by SS. Early after transplantation (10 days),SS could inhibit pancreatic acinar cells from secreting pancreatic amylase and promote islet beta cell proliferation.10 days to 28 days, SS still can inhibit pancreatic acinar cells from secreting pancreatic amylase, but it has no effect on islet beta cell proliferation.SS can reduce pancreas exocrine cells damage in the process of mice islets transplantation.