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The Establishment of Experimental Diabetes Mellitus Rat Model and Indicator Detection of Neuropathy

Author: YinJuan
Tutor: YangHongYing; WangTingHua
Course: Clinical Laboratory Science
Keywords: diabetes mellitus SD rats streptozocin neuropathypou3f3/Brn-1 sciatic nerve diabetic peripheral neuropathygene expression microarray diabetic peripheral neuropathy
CLC: R587.2
Type: Master's thesis
Year: 2012
Downloads: 75
Quote: 0
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Part1:The establishment of experimental diabetes mellitus rat model and indicator detection of neuropathyObjectiveTo establish an experimental type2diabetic rat model,conducting the function and morphology of sciatic nerve to diagnose for the diabetic neuropathy, and prepare for the further investigation.Methods:Sixty male Sprague-Dawley rats were divided in normal diet control group, high fat diet control group and diabetes mellitus group. High fat diet plus streptozotocin was used to induce type2diabetes, detected blood glucose after72hours, observed the successful rate and the mortality rates of streptozotocin-induced diabetes mellitus. Eight weeks after diabetes, nerve conduction velocity is detected by neuroelectrophysiology testing and morphology changes in the sciatic nerve is detected by electron microscopy.Results:Compared to the normal diet control group and the high fat diet group, the level of blood glucose in diabetes mellitus group was significantly higher (P<0.05) after injection of STZ until the experiment ended. And their body weight were increased rapidly before but slowed down to close to the normal diet group after they were injected STZ. Eight weeks after diabetes, part of rats were coincidence with diabetic neuropathy, with sciatic nerve conduction velocity was notably lower (P<0.05), and with apparent pathological changes such as demyelination.Conclusions:The high fat diet combined with small dose STZ can replicate the experimental rat model with progress was similar to type2diabetic of human and the livability is high. The model is conducive to causing vascular neuropathy, it is the ideal animal mode to study the mechanism of type2diabetes and it’s complications.The rats which conduction velocity of sciatic nerve action potential was decreased remarkably by electrophysiology had seriously demyelinating changes in the electron microscope, which indicate the neuroelectrophysiology is a sensitive and specific method of detection of diabetic neuropathy Part2:The change of the expression of pou3f3in diabetic peripheral neuropathyObjectiveTo investigate the change of the expression of pou3f3/Brn-1in mRNA and protein level in diabetic peripheral neuropathy in Streptozotocin-induced rats, to speculate that the physiological and pathological role of pou3f3in diabetic peripheral neuropathy.MethodsAdopted the diabetic rats in part one established, divided into groups according to diabetes mellitus with (DPN) and without (DM) peripheral neuropathy, and the normal diet-fed group (normal control group, NC) and high fat diet-fed group (high-fat control group, HC) as the control, Polymerase Chain Reaction (PCR) is used to detect pou3f3mRNA expression levels, Western Blot (WB) is used to detect pou3f3protein.ResultsThe expression of pou3f3mRNA is decreased in diabetic peripheral neuropathy compared to normal control group (P<0.05), the protein haven’t been detected by Western Blot.Conclusions1. Glucose metabolic disorders may upregulate pou3f3Brn-1mRNA expression and neuropathy may downregulate pou3f3/Brn-1mRNA expression.2. pou3f3protein may not express in sciatic nerves of adult rat. Part3:Gene expression profile analysis in experimental diabetic peripheral neuropathyObjective To acquire the different information of gene expression between rats in experimental DPN, DM and NC group by using gene microarray technology, analyzing the data, and exploring the molecular mechanism of DPN onset so that it may provide assistance for the diagnosis, treatment and prognosis of DPN. Method Adopted the rat models in part one established, extracted total RNA from sciatic nerve of the rats in groups of DPN, DM and NC. CDNA were labeled by RT-PCR, followed microarray hybridization. Using Agilent Rat4x44K Gene Expression Microarrays to detect the expression of gene in sciatic nerves of rats in each group. Scanning images, then data analysis. Results A total of567genes were upregulated and886genes were downregulated in the sciatic nerve of rats in DPN group compared to NC group.532genes were upregulated and693genes were downregulated in the sciatic nerve of rats in DPN group compared to DM group. Conclusion Genes differentially expressed in DPN identified by using cDNA microarray may be provide valuable laboratory bases for clinical diagnosis and therapy of DPN. Key words:gene expression microarray; sciatic nerve; diabetic peripheral neuropathy

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CLC: > Medicine, health > Internal Medicine > Endocrine diseases and metabolic diseases > Islet disease > Diabetic coma and other complications
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