Sphk2 Knockdown Improves STZ-induced Diabetes And Its Renal Complications

Objective:To explore the role of Sphk2 gene knockout on hypoglycemic and reno-protective effect in STZ-induced diabetic mice.Methods:1.The effect of SphK2 gene knockout on blood glucose in diabetic mice induced by STZ.In the study,mice were randomly divided into WT-ND group,Sphk2^-/--ND group,WT-STZ group,Sphk2^-/--STZ group.The model of diabete was induced by STZ,the fasting blood glucose of each group was measured every week.In the experiment,OGTT in each group was detected,as well as ITT and PTT after 8 weeks.blood insulin levels were detected in each group of mice,Immunohistochemical techniques were used to observe the insulin and glucagon in the islet cells,and the tunel method was used to detect apoptosis in the mouse pancreas of each group.2.The effect of SphK2 gene knockout on kidney of diabetic mice induced by STZ.Immunohistochemical technique was used to observe the expression of kidney SphK2 in STZ-induced diabetic nephropathy and spontaneous db/db mice kidney elderly with type 2 diabetes.The SphK2 gene knockout mice were induced diabetic nephropathy by STZ.Before the end of the experiment,24 hours urine was collected and 24 hours of albuminuria was measured in each group.After the experiment,blood was collected from each group of mice to detect serum creatinine and urea nitrogen.The kidneys were stained with H&E,observed pathological morphology and fibrosis of the kidney.To treat diabetic nephropathy mice with SphK2 inhibitor and observe the pathology and FN changes of kidney in mice.Results:1.SphK2 gene knockout effect on diabetic mice induced by STZ.STZ-induced diabetic mice showed a typical symptom of diabete and the"three more and one less"symptoms.Compared with mice in WT-STZ group,after knockout of SphK2gene,it was obvious that STZ-induced diabetic mice were significantly improved by FBG,OGTT,ITT,PTT and enhance blood insulin.Compared with WT-STZ mice,SphK2 gene knockout diabetic mice induced by STZ can relieve the damaged islet cells and apoptosis,In addition,immunohistochemistry was used to observe the increase of insulin and decrease of glucagon in islet cells.2.SphK2 gene knockout the protective effect of STZ-induced kidney in diabetic mice.From immunohistochemistry in paraffin wax film,compared with normal kidney,SphK2 in STZ-induced diabetic nephropathy and spontaneous db/db mice kidney elderly with type 2 diabetes were significantly higher in the kidney expression.And the SphK2^-/--STZ mice had less urine volume than the WT-STZ mice.At the same time,24 hours of albuminuria was detected,and SphK2^-/--STZ mice were less than 24 hours excreted in the WT-STZ mice.there was no difference in serum creatinine between WT-STZ mice and SphK2^-/--STZ mice.To observe the changes of kidney injury in mice,H&E staining showed that pathological change was suppressed in SphK2^-/--STZ mice.PAS and Sirius Red staining observed that reduction ofglycogenandcollagendepositioninSphK2^-/--STZ mice.Immunohistochemistry and q-PCR showed the FN expression in SphK2^-/--STZ mice was reduced.The diabetic nephropathy mice were treated with SphK2 inhibitor for 4 weeks,and the diabetic nephropathy was reduced from H&E,and the expression of FN protein was decreased.Conclusion:After knockout of SphK2 gene,it can improve diabetic blood glucose and reduce islet cells apoptosis.And to alleviate diabetic nephropathy.