【作者】 双卫兵；

【导师】 王东文；

【作者基本信息】 山西医科大学 ， 外科学， 2004， 硕士

【摘要】 目的：探讨糖尿病大鼠膀胱功能的变化情况和糖尿病膀胱功能损伤的病理生理学机制，为临床诊断和治疗糖尿病膀胱提供依据。内容与方法：本研究分为四部分：1、新生鼠腹腔注射大剂量链脲佐素，断乳后再次小剂量追加一次，并应用高糖高脂饮食诱导形成NIDDM 大鼠模型。2、运用离体逼尿肌肌条机械牵张刺激实验结合在体膀胱灌注测压评价NIDDM大鼠在不同时间段的膀胱功能变化。3、在不同时间段，取大鼠膀胱制备匀浆，测定膀胱组织中的SDH活性，并制备逼尿肌电镜标本，观测其线粒体变化。4、制备膀胱匀浆，定磷法测定ATP酶的活性，分离逼尿肌细胞，以Fura-2/AM作为Ca2＋的荧光指示剂，以340nm和380nm为激发波长，510nm为发射波长，进行细胞内钙离子浓度的测定。结果：1.在综合前人经验基础上，运用新的方法成功制备了NIDDM大鼠模型。2.应用机械性牵张刺激试验研究发现：糖尿病发病的最初阶段就出现了逼尿肌兴奋性降低。采用在体膀胱灌注测压来评定糖尿病时膀胱整体功能改变，发现糖尿病对逼尿肌收缩力有损害。3.糖尿病大鼠和正常对照组大鼠膀胱匀浆中SDH活性均有进行性下降趋势，但是糖尿病大鼠下降更为明显，而且在刚发病时变化非常明显。电镜观察到糖尿病大鼠膀胱逼尿肌细胞中线粒体的形态呈现不同程度的肿胀,基质密度下降,线粒体嵴稀疏、变短、断裂甚至消失，线粒体内膜破坏，部分线粒体呈空泡状，这些超微结构改变随发病时间的延长而进行性加重。4.糖尿病大鼠Na-K ATP酶活性下降，依赖ATP供能的Ca-Mg-ATP酶活性也呈下降趋势。但是发病4周（8周组）测定逼尿肌细胞内静息钙离子荧光比值却与正常对照组比较变化不明显。而发病20周（24周组）时，NIDDM组大鼠逼尿肌细胞内钙离子荧光比值比正常对照组明显增高。结论：1.逼尿肌兴奋性降低是DCP功能最早且最明显的改变。2.线粒体破坏导致的逼尿肌能量代谢障碍是引起DCP逼尿肌收缩功能障碍的始动因素。3.在能量代谢紊乱的影响下，细胞膜上的离子泵功能发生异常，并且使得逼尿肌功能发生相应的变化。4.细胞内钙超载可能在进展期DCP逼尿肌功能的损伤的病理过程中起重要作用。更多还原

【Abstract】 Purpose1.To find a kind of methods to manufacture the non-insulin dependent diabetes mellitus rats model.2.To investigate the changes of the NIDDM rats’ detrusor function and provide the data for the clinical urodynamic diagnosis.3.To discover the pathophysiology mechanism of the damage in the NIDDM rats’ detrusor.Material and MethodsThe present study is composed of the following four sub-article: Part one: To manufacture and validate the model rats on non-insulin dependent diabetes mellitus.147 Wistar rats born two days were divided into two groups. The NIDDM group rats underwent intraperitoneal inject with 90mg/kg streptozotocin (STZ). While they have ablaction they had inject with 25mg/kg streptozotocin(STZ) again. Then these rats were fed with the diets enriched with sucrose (20% w/w) and lard(18%.w/w)and yolk powder(3%,w/w).Another group served as controls. Part two: Experimental study of excitability and autorhthmicity and contraction /relaxation in urinary bladder of NIDDM rats The NIDDM group rats and controls group rats had been examined by urodynamic testing machine after they have been fed special diats12th and 24th weeks. At 4th/8th/12th/16th/20th/24th weeks they had been fed special diets, the diabetes rats and controls rats tested by detrusor strips in vitro. Part three: Experimental study of succinate dehudrogenase change and ultra-structural change of mitochondrion of detrusor in diabetes ratswhile they have ablaction and after 4th/8th/12ve/16th/20th/24th weeks they had been fed special diets, the diabetes rats and controls rats had been killed ,and <WP=6>the bladder make into homogenate. Determined the activity of the succinate dehydronase by chromatometry . The ultrastructures of mitochondrion in detrusor cell of each rat were examined after the 4th/12ve/24th week they had been fed the special diets.Part four: The changes of adenosine-triphosphatase activity in bladder homogenate and cytosol-free Calcium concentration in single bladder smooth muscle cells from normal and diabetic rats.The adenosine-triphosphatase activity had been examinated by chromatometry. The effect of diabetes mellitus on intracellular calcium concentration was measured in single rat bladder smooth muscle cells using the fluorescent calcium indicator dye Fura-2/AM at rest situation.Result 1.The model on non-insulin-dependent diabetes mellitus is successful.2.At the early stage, the excitability of diabetes rats was depressed. The contraction and relaxation of diabetes rats were lower than the rats in controls group. The autorhthmicity of the diabetes rats was increased before 22nd weeks, then decreased than the controls rats. The bladder weight /residual urine and bladder compliance were increased on diabetes rats. 3.The activity of the succinate dehydronase (SDH) decreased at the in the initial stage of NIDDM rats. The damage of mitochondrion’s ultrastructures in detrusor cell became manifest by progression.4. The adenosine-triphosphatase activity had been decreased at the initial stage by the effect of diabetes mellitus. The intracellular calcium concentration in single rat bladder smooth muscle cells had no significant difference in the initial stage at rest situation. After 20 weeks they fall ill, The intracellular calcium concentration in single rat bladder smooth muscle cells had increased than control rat’s.Conclusion 1.The excitability depressed is the first change in diabetes rats’ detrusor. The autorhthmicity had also changed at the early stage .The damage of contraction <WP=7>and the relaxation maybe the reasons of the compliance change on the diabetes rats’ bladder.2.Detrusor contraction was impaired in the early stage of diabetes. The mitochondrion destroyed and the detrusor metabolism dysfunction should been one of the reasons. 3.Intracellular calcium overload occurred latter than the dysbolism.4.To control the dysbolism and use ATP or the calcium antagon maybe a good treatment of the diabetic cystopathy.更多还原