越婢汤治疗遗尿症逼尿肌不稳定的机理探讨

发布时间：2018-06-07 16:00

本文选题：遗尿症 + 逼尿肌不稳定　；参考：《成都中医药大学》2014年博士论文

【摘要】：研究背景：遗尿症是儿童临床常见疾病之一。遗尿患儿长期得不到诊治,将会降低其生活质量,影响儿童心理和人格的健康发展,其心理问题甚至会遗留终身。由此,探寻遗尿症早期而有效的干预,是一项具有深远社会意义的工作,也是一个重大而艰巨的课题。近年来,现代医学对本病展开了多方位的研究,其具体发病机制虽不十分明确,但膀胱逼尿肌不稳定无疑已成为目前国内外研究本病的主要方向之一,逼尿肌不稳定是临床最常见的膀胱尿道功能障碍之一,而膀胱中ICCs样细胞数量的增多可能是引起逼尿肌不稳定产生的主要原因之一；一氧化氮(NO)作为抑制性非肾上腺能神经非胆碱能神经(NANC)神经递质,也参与了膀胱功能的调节。同时,作为目前为止研究得最清楚的神经营养因子,神经生长因子(NGF)水平的变化可直接影响病理性膀胱神经的分布。中医对本病的认识有两千多年的历史,积累了丰富的临床治疗经验。因此从中医药研究本病有可能成为在本病治疗方法上的有益补充。我们在前期的临床观察中发现越婢汤治疗小儿遗尿症疗效肯定,故本研究以越婢汤为干预方剂,探讨越婢汤对遗尿症不稳定膀胱模型的作用机制。研究目的：探讨越婢汤对遗尿症不稳定膀胱模型的作用机制,从而对中医“提壶揭盖”“下病上取”,以及“肺为水之上源”等理论作出现代实验科学的严谨论证。研究方法：将108只wistar大鼠随机分为膀胱出口梗阻手术组和假手术组,6周后行尿动力检查筛选出手术组膀胱逼尿肌不稳定大鼠和假手术组膀胱逼尿肌稳定大鼠进行研究。将手术组膀胱逼尿肌不稳定大鼠随机分为越婢汤古方组、越婢汤高剂量组、越婢汤中剂量组、越婢汤低剂量组、缩泉丸对照组、模型对照组,加上假手术组共7组,每组10-13只大鼠。给药14天后观察越婢汤各剂量组对模型的影响。从膀胱湿重、膀胱脓肿和结石情况宏观观察药物对模型的疗效,用免疫组化和western blot方法测试膀胱ICCS细胞含量；用酶联免疫法(ELISA).免疫组化测试血清和膀胱NOS蛋白含量；用ELISA、免疫组化测试血清、下丘脑和膀胱NGF蛋白含量；并做出数据处理分析,从多角度探讨越婢汤对不稳定膀胱的可能作用机制。研究结果： 1.造模结果：手术6周后,经尿动力学检测仪进行充盈性膀胱测压检测,假手术组大鼠膀胱逼尿肌不稳定(DI)发生率为16.7%(2/12)；手术组96只,尿路感染、漏尿死亡15只,其中63只出现DI,发生率为77.8%(63/81)。 2.宏观疗效：模型对照组大鼠膀胱发生膀胱脓肿1只,膀胱结石2只；越婢汤低剂量组大鼠发生膀胱脓肿1只,膀胱结石1只；其余各组未发现明显膀胱脓肿和膀胱结石。与假手术组相比,模型对照组大鼠膀胱湿重明显增高,具有非常显著的统计学意义(P0.01),与模型对照组相比,越婢汤组大鼠膀胱湿重明显减轻,具有统计学意义(P0.05)。 3.药物对膀胱ICCs细胞的影响：与假手术组比较,模型组膀胱逼尿肌ICCs细胞显著增多(P0.05)；与模型组比较,越婢汤干预后膀胱逼尿肌ICCs细胞有显著减少(P0.05)或表现出一定的量效关系。 4.药物对NOS的影响： (1)膀胱组织免疫组化检查：与假手术组相比,模型对照组大鼠膀胱NOS含量明显增多,具有非常显著的统计学意义(P0.01)；与模型对照组相比,越婢汤干预后大鼠膀胱NOS含量明显减少,具有显著的统计学意义(P0.01),提示：越婢汤可以明显下调大鼠膀胱NOS含量。 (2)血清ELISA检测：与假手术组相比,模型对照组大鼠血清NOS含量明显增多,具有非常显著的统计学意义(P0.01)；与模型对照组相比,越婢汤干预后大鼠血清NOS有下调趋势。 5.药物对神经生长因子(NGF)的影响： (1)膀胱组织免疫组化检查：与假手术组相比,模型对照组大鼠膀胱中NGF蛋白含量明显降低,具有非常显著的差异(P0.01)；与模型对照组相比,越婢汤干预后大鼠膀胱中NGF蛋白含量明显升高,具有显著的差异(P0.05),说明越婢汤具有上调DI大鼠膀胱组织NGF的作用。 (2)下丘脑免疫组化检查：与假手术组相比,模型对照组大鼠下丘脑中NGF蛋白含量明显降低,具有非常显著的差异(P0.01)；与模型对照组相比,越婢汤干预后大鼠下丘脑中NGF蛋白含量明显升高,具有非常显著的差异(P0.05)；说明越婢汤具有上调DI大鼠下丘脑NGF的作用。 (3)血清ELISA检测：与假手术组相比,模型对照组大鼠血清中NGF蛋白含量明显降低,具有非常显著差异(P0.01)；与模型对照组相比,越婢汤干预后大鼠血清中NGF蛋白含量明显升高(P0.05),说明越婢汤具有上调DI大鼠血清NGF的作用。研究结论： 1.越婢汤可以显著改善逼尿肌不稳定引起的膀胱功能障碍。 2.逼尿肌不稳定膀胱中ICCs细胞数量升高；膀胱组织和血清NOS含量升高；血清、下丘脑和膀胱组织中NGF含量均降低。 3.越婢汤古方组和高剂量组可以明显下调膀胱ICCs细胞数量,下调膀胱组织NOS含量,上调血清、下丘脑和膀胱组织中NGF含量,其对遗尿症的治疗机理可能与此有关。
[Abstract]:Research background:
Enuresis is one of the common clinical diseases in children. Children with enuresis can not be diagnosed and treated for a long time, which will reduce their quality of life, affect the healthy development of children's psychology and personality, and their psychological problems will remain for life. Therefore, to explore the early and effective intervention of enuresis is a work with far-reaching social significance and a great and hard work. In recent years, modern medicine has carried out a multidimensional study on this disease. Although its specific pathogenesis is not very clear, the detrusor instability of the bladder has undoubtedly become one of the main directions for studying this disease at home and abroad. Detrusor instability is one of the most common bladder urethral dysfunction, and ICCs like cells in the bladder. The increase in quantity may be one of the major causes of detrusor instability; nitric oxide (NO), as an inhibitory non adrenalenine non cholinergic neurotransmitter (NANC), is also involved in the regulation of bladder function. At the same time, as the most clearly known neurotrophic factor, the level of Neurogrowth factor (NGF) is at present. Changes can directly affect the distribution of pathological bladder nerves.
Chinese medicine has a history of more than two thousand years and has accumulated rich experience in clinical treatment. Therefore, the study of this disease from traditional Chinese medicine may be a useful supplement to the treatment of this disease. Effect of Yue Mau Decoction on unstable bladder model of enuresis.
The purpose of the study is:
The mechanism of Yue maidu Decoction on the unstable bladder model of enuresis is discussed, and the rigorous demonstration of modern experimental science is made on the theory of "lifting the lid of the pot" and "taking the disease on the bottom" and "the lung is the source of water" in traditional Chinese medicine.
Research methods:
108 Wistar rats were randomly divided into the operation group of bladder outlet obstruction and the sham operation group. After 6 weeks, the urinary bladder detrusor instability rats and the sham group were selected to study the bladder detrusor stable rats. The bladder detrusor instability rats were randomly divided into the Yue maidu Decoction group and the high dose of the Yue Maidan Decoction group. The dose group of Yue maidu decoction, the lower dose group of Yue maidu, the control group of the shinquan pill, the model control group, and the sham operation group, 7 groups, each group of 10-13 rats. After 14 days of administration, the effect of each dose group on the Yue Maidan Decoction on the model was observed. The effect of the drug on the model was observed from the wet weight of the bladder, bladder abscess and stone, by immunohistochemistry and Western blot prescription. The content of ICCS cells in bladder was tested by method of enzyme linked immunosorbent assay (ELISA). The content of serum and bladder NOS protein was measured by immunohistochemistry; the content of NGF protein in the hypothalamus and bladder was measured with ELISA, immunohistochemistry and the data processing analysis was made to discuss the possible mechanism of the action of Yue maidu soup to the unstable bladder.
The results of the study:
1. model results: after 6 weeks of operation, the bladder detrusor instability (DI) was 16.7% (2/12) in the sham operation group, 96 in the operation group, 15 in urinary tract infection and leakage of urine, of which 63 had DI, and the incidence was 77.8% (63/81).
2. macroscopical effect: the bladder abscess in the bladder of the model control group was 1, and the bladder stone was 2. The lower dose group of Yue maidu decoction had 1 bladder abscesses and 1 bladder stones. The other groups did not find obvious bladder abscess and bladder stone. Compared with the sham operation group, the wet weight of the bladder was significantly higher in the model control group, which was very significant. Statistical significance (P0.01), compared with the model control group, the weight of bladder wet weight of the rats in the Yue maiden group was significantly reduced, with statistical significance (P0.05).
The effect of 3. drugs on bladder ICCs cells: compared with the sham group, the ICCs cells of the bladder detrusor muscle in the model group increased significantly (P0.05). Compared with the model group, the ICCs cell of the bladder detrusor was significantly reduced (P0.05) or showed a certain dose effect relationship with the model group.
4. effects of drugs on NOS:
(1) immunohistochemical staining of bladder tissue:
Compared with the sham operation group, the NOS content of the bladder in the model control group increased significantly, and had significant statistical significance (P0.01). Compared with the model control group, the NOS content of the bladder in the rats after the Yue Maidan Decoction decreased significantly (P0.01). It was suggested that the Yue Maidan decoction could obviously reduce the content of NOS in the bladder of rats.
(2) serum ELISA detection:
Compared with the sham operation group, the serum NOS content of the rats in the model control group increased significantly and had significant statistical significance (P0.01). Compared with the model control group, the serum NOS of the rats of the Yue Maidan decoction was downregulated.
5. effects of drugs on nerve growth factor (NGF):
(1) immunohistochemical staining of bladder tissue: compared with the sham group, the NGF protein content in the bladder of the model control group was significantly lower than that in the model control group (P0.01). Compared with the model control group, the content of NGF protein in the bladder of the Yue maidu decoction was significantly higher than that in the model control group, with significant difference (P0.05), indicating that the Yue maidu soup was up to up DI. The effect of NGF on the bladder tissue of rats.
(2) immunohistochemical examination of hypothalamus: compared with the sham group, the NGF protein content in the hypothalamus of the model control group was significantly lower than that in the model control group (P0.01). Compared with the model control group, the NGF protein content in the hypothalamus of the Yue maidu decoction was significantly higher than that in the model control group, with a very significant difference (P0.05). It can increase the effect of NGF in the hypothalamus of DI rats.
(3) serum ELISA detection: compared with the sham group, the serum NGF protein content in the model control group was significantly lower than that in the model control group (P0.01). Compared with the model control group, the serum NGF protein content in the serum of the Yue Maidan decoction was significantly increased (P0.05), and that the Ming Yue maidservant soup was able to increase the role of NGF in the serum of DI rats.
The conclusions are as follows:
1. Yue maidu decoction can significantly improve bladder dysfunction caused by detrusor instability.
2. detrusor instability, ICCs cells in bladder increased, bladder tissue and serum NOS content increased, serum, hypothalamus and bladder tissue NGF content decreased.
The number of bladder ICCs cells and the content of NOS in bladder tissue were down regulated by the 3. Yue maidservant Decoction and the high dose group, and the content of NGF in the serum, hypothalamus and bladder tissue was up-regulated. The mechanism of the treatment of enuresis may be related to this.
【学位授予单位】：成都中医药大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R272

【参考文献】