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Objective To investigate the improvement and effect of the method of islet extraction in mice. Methods According to different islet extraction methods, all mice were randomly divided into the common bile duct puncture group (n=100) and common bile duct puncture combined with in situ pancreatic injection group (combined injection group, n=100). Common bile duct puncture combined with in situ pancreatic injection was utilized as the modified method. The islets were selected and purified under stereomicroscope. The morphology and purification of islets were identified. The islet yield and success rate of islet extraction were statistically compared between two groups. The survival of islets after 1 week culture in vitro was analyzed, and the insulin secretion function of islets after 24 h and 4 d culture in vitro was evaluated. Results Compared with the common bile duct puncture group, the islet yield in the combined injection group was significantly increased (P < 0.001). The success rate of islet extraction in both groups was 83% with no statistical significance (P > 0.05). The islets extracted by common bile duct puncture combined with in situ pancreatic injection had intact morphology, high purity and high activity. The survival rate of newly isolated islets was nearly 100% after 24 h culture in vitro. After 1~5 d culture in vitro, the islet cells survived well. After 6 d culture in vitro, the islets showed central death. After culture in vitro for 24 h and 4 d, the islet function of the mice was normal after high glucose stimulation. Conclusions Common bile duct puncture combined with in situ pancreatic injection can increase the islet yield, and the obtained islet cells have high activity and proper function.
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Objective To investigate the role of glargine in glucose metabolism improvement and antiinflammation of skeletal muscle in Caveolin-1 silenced type 2 diabetic mice.Methods Multiple low doses of streptozotocin (STZ) intraperitoneal injection and high-fat high-glucose (HFHG) were used to induce type 2 diabetic mice model.The mice were divided into normal control group (NC group) and type 2 diabetic model group (T group).Then according to virus injection and glargine treatment,T group were further divided into type 2 diabetes group (T2DM group),type 2 diabetes with insulin treatment group (insulin group),Caveolin-1 silenced with insulin treatment group (LV-CAV1 group),and scramble virus with insulin treatment group (LV-GFP group).Glucose metabolism was accessed by the fluctuation of blood glucose.TNF-α protein expression in skeletal muscle was detected by Western blot.Results The glycemic control of LV-CAV1 group needed more dosages of glargine (P ＜ 0.05).The expression of TNFαin skeletal muscle was elevated in LV-CAV 1 group than that in LV-GFP and insulin group (P ＜ 0.05).Conclusion The anti-inflammation function and glycemic metabolism improvement of glargine may be associated with the expression of Caveoin-1 in skeletal muscle.