| Purpose:Type Ⅰ diabetes is due to the complete destruction of pancreatic beta cells by immune attack,resulting in insufficient insulin secretion.In addition to supplementing exogenous insulin,islet transplantation is an effective treatment for type 1 diabetes.A suitable islet transplantation site can promote islet survival and improve the success rate of islet transplantation.Therefore,based on the knowledge of retroperitoneal anatomy,this thesis performed retroperitoneal islet transplantation and found that retroperitoneal islet transplantation was superior to renal capsule islet transplantation.Methods:（1）Islet extraction and purification: 8-week-old Balb/c male mice were used as the donor and recipient of islet transplantation.The pancreatic islets were extracted by puncturing the common bile duct and filling the pancreas with collagenase P,and the islets were purified by Lympholyte 1.1 density gradient centrifugation.（2）Modeling: The Balb/c mouse type Ⅰ diabetes model was induced by streptozotocin.On the third day of modeling,the blood glucose reached 324 mg/d L and above,which was regarded as successful modeling.（3）Grouping: The model mice were randomly divided into four groups,renal capsule control group,peritoneal experimental group,peritoneal sham operation group and STZ group.In order to control irrelevant variables,the islet equivalent was calculated,and the number of islets transplanted was unified.At the same time,no immunosuppressant was used in all transplant groups.（4）Operation: Transplantation was performed,the operation complexity and operation time of the renal capsule control group and the peritoneal experimental group were compared,and the peritoneal anatomy was observed.（5）Transplantation effect: random blood glucose observation was performed on the transplanted mice to evaluate the survival curve.On the 7th day and the 14 th day of transplantation,the mice were slowly injected with 25% glucose 100 μL through the tail vein,and the glucose tolerance test was performed.Blood glucose concentration was measured at 0,5,15,30,60,90,and 120 minutes.The renal capsule and peritoneal grafts were harvested and embedded in paraffin to observe the immunofluorescence of insulin,and the insulin secretion ability of the transplanted islets between the two groups on the7 th day and the 14 th day was compared.Results:（1）Islet preparation: The islets were digested with collagenase P,and the islets of an8-week-old Balb/c mouse were extracted by Lympholyte 1.1 separation and purification.（2）Surgery: Compared with the renal capsule control group,the retroperitoneal experimental group had less time for transplantation.（3）Survival curve: Compared with the renal capsule control group,the survival percentage of the retroperitoneal experimental group was higher.（4）Random blood sugar: Compared with the peritoneal sham operation group and the STZ group,the random blood sugar in the retroperitoneal experimental group and the renal capsule control group was significantly reversed on the14 th day after transplantation.Compared with the renal capsule control group,the random blood glucose of the retroperitoneal experimental group was significantly different（p<0.01）,and the retroperitoneal experimental group was better than the renal capsule control group.On the 100 th day after transplantation,the blood glucose of the experimental group was less than 200 mg/d L in the retroperitoneum,which was better than that of the renal capsule control group.（5）Glucose tolerance: On the 7th day of transplantation,the blood glucose concentration in the retroperitoneal experimental group first dropped below 200 mg/d L within 120 minutes.On the 14 th day of transplantation,the blood glucose of both the retroperitoneal experimental group and the renal capsule control group dropped below 200 mg/d L within 120 minutes,and the blood glucose concentration of the retroperitoneal experimental group dropped to below 200 mg/d L preferentially compared with the renal capsule control group.（6）Immunofluorescence:There was a statistically significant difference in the mean immunofluorescence intensity of insulin between the retroperitoneal experimental group and the renal capsule control group on the 7th day after transplantation（p<0.05）.The insulin secretion of the retroperitoneal experimental group was better than that of the renal capsule control group.Conclusion:（1）Pancreatic islet transplantation can stabilize blood glucose in type 1 diabetic mice for a long time,which is an effective way to treat type 1 diabetes.（2）The retroperitoneum has abundant blood supply,which can provide sufficient nutrition for islets,and can be used as the dominant part of islet transplantation.After comparing random blood glucose,glucose tolerance,and insulin secretion immunofluorescence,retroperitoneal islet transplantation was found to be superior to renal capsule islet transplantation. |
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