三型酸敏感离子通道与大鼠膀胱逼尿肌活动关系的研究

发布时间：2018-06-12 05:42

本文选题：尿流动力学 + 阿米洛利　；参考：《昆明医科大学》2014年硕士论文

【摘要】：目的：探讨大鼠膀胱组织中三型酸敏感离子通道(acid-sensing ion channels,ASIC3)表达的变化及与膀胱逼尿肌活动的关系,明确三型酸敏感离子通道蛋白与尿路刺激症的关系。为临床尿路刺激征以及OAB的治疗提供新的研究思路。方法：选取成年雌性大鼠,通过环磷酰胺(CYP)腹腔注射、大肠杆菌膀胱灌注方法构建大鼠OAB模型及细菌性膀膀胱炎模型。根据造模方法的不同,分为1、CYP大组,2、细菌大组。每组再分为正常组、OAB组(CYP大组)或膀胱炎组(细菌大组)、干预组。即：A1：正常组、B1:OAB组、C1:OAB干预组(OAB+阿米洛利)；A2:正常组、B2:膀胱炎组、C2：膀胱炎干预组(膀胱炎+阿米洛利)。对六组大鼠膀胱灌注生理盐水或阿米洛利生理盐水溶液,进行尿流动力学检测。同时取六组大鼠的膀胱组织,进行病理学检查。并通过免疫组织化学染色、RT-PCR和western-blot方法测量其ASIC3含量的差异。结果：尿流动力学提示：正常组大鼠储尿期及排尿期未见明显不稳定收缩；OAB组、膀胱炎组及两干预组在储尿期可见不稳定收缩,与正常组相比：排尿间隔明显缩短(P0.01),排尿次数明显增加(P0.01)；OAB干预组与OAB组相比,不稳定收缩次数较少,排尿次数减少、排尿间隔延长(P0.05)；膀胱炎干预组与膀胱炎组相比,不稳定收缩次数较少,排尿次数减少、排尿间隔延长(P0.05) 病理学检测提示OAB组、膀胱炎组、OAB干预组及膀胱炎干预组中大鼠膀胱粘膜破坏；免疫组化提示：大鼠膀胱组织含有ASIC3;且ASIC3主要存在于膀胱粘膜上,ASIC抑制剂可以抑制膀胱粘膜上ASIC3的表达；肌层中含有少量的ASIC3。ASIC3抑制剂对肌层ASIC3无明显作用。 RT-PCR及western-blot提示：OAB组及膀胱炎组中膀胱上ASIC3表达明显升高(P0.01),加入ASIC3抑制剂后,OAB干预组、膀胱炎干预组ASIC3表达有所降低(高于正常组,P0.05；低于OAB组或膀胱炎组,P0.05)。结论： 1、大鼠膀胱组织内有ASIC3的基因及蛋白表达,且ASIC3的基因及蛋白表达主要存在于膀胱粘膜上； 2、OAB大鼠模型和膀胱炎大鼠膀胱组织上ASIC3的基因及蛋白表达较正常大鼠增高,其表达量受ASIC抑制剂的影响较为明显。 3、膀胱内ASIC3表达升高膀胱逼尿肌反射亢进密切相关。 4、阿米洛利可能成为潜在的治疗膀胱逼尿肌反射亢进的药物
[Abstract]:Objective: to investigate the expression of acid-sensing ion channel ASIC3 in rat bladder and its relationship with detrusor activity, and to clarify the relationship between acid sensitive ion channel protein and urinary tract irritation. Methods: OAB model and bacterial bladder cystitis model of adult female rats were established by intraperitoneal injection of cyclophosphamide (Cyp) and intravesical instillation of Escherichia coli. According to the different methods of modeling, it was divided into 1 CYP group (2) and bacteria group (2). Each group was divided into normal group (OAB group) or cystitis group (large bacterial group, intervention group). That is: normal group: B 1: OAB group C 1: OAB intervention group A 2: normal group B 2: cystitis group C 2: cystitis intervention group C 2: cystitis amilolol group. Six groups of rats were given intravesical instillation of normal saline or amiloride saline solution, and the urodynamics was measured. At the same time, the bladder tissues of six groups of rats were taken for pathological examination. The content of ASIC3 was measured by immunohistochemical staining RT-PCR and western-blot. Results: the urodynamics indicated that there was no obvious unstable contraction in normal rats during the period of urine storage and urination. In the cystitis group and the two intervention groups, the unstable contraction was observed in the period of urinary storage. Compared with the normal group, the interval of urination was significantly shorter than that in the control group, and the frequency of urination increased significantly in the OAB group compared with the OAB group, the frequency of unstable contraction was less, and the frequency of urination was less in the OAB group than in the OAB group. Compared with cystitis group, the number of unstable contraction was less and the number of micturition decreased in cystitis intervention group. The bladder mucosa was destroyed in the OAB group and the cystitis intervention group, and the expression of ASIC3 was inhibited by ASIC3 in the bladder tissue of the rats, and ASIC3 was mainly present in the bladder mucosa of the rats, and the expression of ASIC3 in the bladder mucosa was inhibited by ASIC3 inhibitor. ASIC3. ASIC3 inhibitor in myometrium had no obvious effect on ASIC3. RT-PCR and western-blot indicated that ASIC3 expression in bladder increased significantly in the two groups, and the expression of ASIC3 in the bladder was significantly increased after adding ASIC3 inhibitor, and the expression of ASIC3 was significantly increased in the control group after addition of ASIC3 inhibitor, and the expression of ASIC3 was significantly increased in the control group after addition of ASIC3 inhibitor. The expression of ASIC3 in cystitis intervention group was lower than that in normal group (P 0.05), and lower than that in OAB group or cystitis group (P 0.05). Conclusion: 1, ASIC3 gene and protein expression were found in bladder tissue of rats. The expression of ASIC3 gene and protein was mainly found in bladder mucosa, and the expression of ASIC3 gene and protein in bladder tissue of rats with OAB and cystitis was higher than that of normal rats. The expression of ASIC3 was significantly affected by ASIC inhibitor. 3The increase of ASIC3 expression in bladder was closely related to hyperreactivity of bladder detrusor reflex. 4. Amilolol may be a potential drug for the treatment of bladder detrusor hyperreflexia.
【学位授予单位】：昆明医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R694.5

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