摘要
背景：慢性前列腺炎(Chronic prostatitis, CP)是泌尿外科常见病、多发病,其病因、病理改变和临床症状复杂多样,发病机制不明,其中自身免疫反应可能起着重要的作用。排尿功能障碍是CP主要临床表现之一,目前认为功能性后尿道梗阻是CP排尿功能障碍的主要原因。梗阻可诱发逼尿肌无抑制收缩,形成不稳定膀胱。部分CP患者有膀胱的尿动力学异常改变,但对其产生的机制相关的研究相对较少。
膀胱逼尿肌中存在的ICC样细胞与神经末梢关系密切,具有起搏功能,研究发现不稳定膀胱中ICC样细胞的数量及存在形式都发生了改变。膀胱功能异常是排尿功能障碍的常见原因,那么慢性前列腺炎的膀胱ICC样细胞的数量及存在形式都发生了怎样的改变,以及这种改变与膀胱功能异常之间的关系如何?值得深入研究。
青藤碱(SIN)有抗炎和免疫抑制以及对胃肠道平滑肌的抑制作用,而应用于临床；针灸治疗慢性前列腺炎及抑制膀胱活动亢进的作用也有明确报道。但其作用机理鲜有报道。
目的：观察慢性非细菌性前列腺炎大鼠模型膀胱功能及膀胱ICC样细胞网络的变化,研究中药青藤碱和针刺治疗对慢性前列腺炎及膀胱功能的影响,探讨青藤碱和针刺治疗慢性前列腺炎的机理。为慢性前列腺炎及其膀胱功能障碍的治疗提供新的思路。
方法：采用同种系SD大鼠前列腺蛋白提纯液,辅以双重免疫佐剂方法,建立实验性慢性非细菌性前列腺炎(CAP)大鼠动物模型。将动物随机分为模型对照组、针刺组、青藤碱大、中、小剂量组五组。青藤碱大、中、小剂量组分别为30mg/kg、20 mg/kg和10 mg/kg体重,腹腔注射,每日一次,连续14天。针刺组电针中膂俞、会阳,电针治疗参数：输出电压4～6 V,疏波4 Hz,密波20 Hz,留针20 min。1次/d,共治疗10d。治疗后尿流动力学检测描记各组膀胱内压力图,取前列腺组织,观察其病理改变,逼尿肌组织铺片,免疫荧光检测c-kit阳性细胞的表达。用用双抗体夹心.酶联免疫吸附试验(ABC-ELISA)法测定前列腺组织匀浆IL-1β、PGE2和β-EP的含量。RT-PCR检测膀胱逼尿肌中NGFmRNA的表达。
结果：与正常大鼠相比,模型组动物充盈性膀胱压力测定显示膀胱最大容量和最大逼尿肌压明显减小,残余尿量明显增加,70%的大鼠在膀胱充盈期出现程度不等的逼尿肌不稳定性收缩。模型组前列腺病理符合慢性前列腺炎改变,证实CAP可以引起膀胱功能的改变,为前列腺炎膀胱功能异常提供了功能学上的依据。但模型组慢性前列腺炎各病理分级中不稳定膀胱比率无明显差别。
与正常对照组相比,模型组Kit阳性的ICC样细胞数量增多,积分光密度值(IOD)降低,部分ICC样细胞排列紊乱,细胞间的联系散乱
与模型组相比,针刺会阳、中膂俞和青藤碱治疗后,发生逼尿肌不稳定收缩的动物数量明显减少,程度减轻。
青藤碱大剂量组和针刺组大鼠前列腺上皮组织恢复正常,管壁结构完整,间质水肿消失。青藤碱中剂量组和小剂量组对实验大鼠前列腺组织结构的破坏虽然也有一定程度的改善,但作用较轻。
与模型组相比,不同剂量青藤碱治疗组随青藤碱使用剂量增加,ICC样细胞数量逐渐减少,IOD逐渐增高,以青藤碱大剂量组最接近正常对照组。
模型组NGF mRNA表达明显升高,各青藤碱治疗组随青藤碱剂量增加NGF mRNA表达依次降低,针刺组和青藤碱大剂量组NGF mRNA表达接近正常组。各组逼尿肌组织NGF的改变与ICC样细胞的变化呈现一致性。
青藤碱大、中、小剂量组及针刺组大鼠前列腺组织内PGE2及IL-1β的水平较模型组明显降低；β-EP的水平较模型组明显升高。青藤碱大剂量组、针刺组大鼠前列腺组织匀浆中IL-1β含量与正常组比较,差异无统计学意义(P>0.05)
结论：采用自身免疫反应所造成的CAP动物模型,其病理改变与临床的病理改变较为接近,是一种较为理想的的造模方式。
慢性前列腺炎模型大鼠可以出现以逼尿肌不稳定收缩为主的膀胱功能异常,以及膀胱ICC样细胞分布和数量的异常。膀胱ICC样细胞结构和功能的变化可能与CP膀胱功能异常的产生有关。
盐酸青藤碱和针刺会阳、中膂俞能够降低大鼠前列腺组织内PGE2及IL-1β的水平,升高p-EP的水平,可减轻前列腺组织的炎症反应,改善前列腺组织结构的破坏,对前列腺组织起到一定的保护作用。
针刺会阳、中膂俞和青藤碱对前列腺组织的保护作用可以影响膀胱ICC样细胞的数量和分布,减轻继发的膀胱的肌源性损害,从而调节膀胱功能。针刺和青藤碱对慢性前列腺炎膀胱NGF产生抑制作用,可以从某种程度上说明膀胱ICC样细胞与NGF存在交互作用,提示,针刺和青藤碱可能通过免疫抑制和对细胞因子和神经递质的调节等作用参与到膀胱ICC样细胞网络的调节。
Background:Chronic non-bacterial prostatitis/chronic pelvic pain syndrome (CP/CPPS) is a common disease of male reproductive system. The etiology and the pathogenesis of CP are not yet clear so far. Because of the high incidence of the disease, lack of typical symptoms, and difficulty to treat, CP seriously impairs the quality of life in patients. Recently studies have shown that CP is the result of a variety of factors. Autoimmune reactions play an important role in the pathogenesis which is mediated by inflammatory cytokines. Pain, voiding dysfunction and sexual dysfunction are three main manifestations of CP. Many investigations ascribed the voiding dysfunction to the functional obstruction at posterior urethra which evokes non-inhibitive contraction and eventually leads to unstable bladder. However, among some patients without functional urethral obstruction, symptoms and abnormal urodynamic findings are also present.
Interstitial cells of Cajal (ICC) in the bladder act as pacemaker and have strong relationship with nerve terminal. The increase of ICC-like cells in the detrusor, which maybe lead to an enhanced spontaneous contraction of detrusor, and may be one of the important mechanisms of detrusor instability (DI). The abnormal function of bladder was the main reason for the voiding dysfunction. The question is:are there any relationship with the alteration of bladder ICC-like cells in the association between voiding dysfunction and CP? To the best of our knowledge, there were very few studies addressing this question.
Sinomenine was widely used in clinical practice because of its effects of anti-inflammatory, immunosuppression and inhibitory action to gastro-intestinal smooth muscle. It was recognized that acupuncture on relative points had therapeutic effects on CP symptoms and bladder hyperactivity. As such, elucidation of the mechanism at myogenic level of detrusor of bladder will facilitate treatment of CP and the concomitant bladder dysfunction.
Objective:To observe the alteration of bladder function and its ICC-like cells in SD rat model of experimental autoimmune prostatitis (EAP), and to investigate the effects of Sinomenine and acupuncture at Zhonglushu and Huiyang point on morphological peculiarities of experimental rats prostate tissue and the concomitant bladder dysfunction, and evaluated their efficacy.
Method:The model of experimental autoimmune prostatitis (EAP) rat was made by DPT vaccine in peritoneal injection and rat prostatein purification of FCA emulsion multiple intra-dermal injections for 45 days. Cystometries were performed in conscious rats after suprapubic bladder puncture. The bladder capacity, pressure and postvoiding volume were measured and the incidence of detrusor instability was studied. Then prostate glands were obtained to investigate the histomorphological changes. Detrusor tissue pieces were obtained for detecting c-kit positive cells by immunofluorescent staining.
Fifty SD rats were randomly divided into a control group, a model group, an acupuncture group and the groups of low, medium and high dose of Sinomenine. The low, medium and high dose group was given dose of 10, 20,30mg/kg, respectively, by intraperitoneal injection. The acupuncture group was electroacupunctured at Huiyang (BL 35) and Zhonglushu (BL 29) with output voltage of 4～6V, dense wave of 20 Hz and distant wave of 4 Hz. The above groups were subsequently treated for 14 days, qd. Finally, cystometries, histomorphological observation of prostate and detection of c-kit positive cells in bladder were performed. The bladder detursor tissues were harvested for a reverse transcription polymerase chain reaction (RT-PCR) exam of NGFmRNA.The contents of IL-1β, PGE2 and β-endorphin in prostate-homogenate were detected by enzyme linked immunosorbent assay.
Results:The incidence of DI developed in model group was 70%. Bladder pressure and capacity decreased from (27.3±1.87) cmH2O and (0.57±0.06) mL in control rats to (22.8±2.03)cmH2O and (0.39±0.05)mL in model rats, respectively (P< 0.05). The postvoiding volume increased from (0.11±0.04)mL to (0.33±0.06)mL. The model group showed chronic inflammation in prostate glandular tissue. The above results indicated that the model was built successfully and was a candidate for further study on bladder dysfunction secondary to CP.
Compared with control group, model group showed considerably increase in number of ICC-like cells and decrease of integral optical density value (IOD) in the detrusor. The ICC-like cells were arranged disordered, and the intercellular connection was scattered in model group.
After acupuncture and administration of Sinomenine, DI ratio of model group, acupuncture group, Sinomenine high dose group, medium dose group and low dose group was 60%,10%,20%,40%, and 50%, respectively. Compared with that of the model group,the bladder capacity and maximum detrusor pressure increased,and the residual volume decreased significantly in the treatment group.
In the high dose group of Sinomenine and acupuncture group, the epithelial tissues recover, the architecture of epithelial tissues were integrated, and the interstitial edemas disappeared. In the mid and low dose group, the damage of prostate gland tissues recovered slightly. According to classified statistical analysis, the differences among each group of the Sinomenine and the acupuncture group had statistical significance.
Compared with the model group, the number of ICC-like cells (c-kit positive cells) decreased significantly and ICC-like cells were arranged more orderly in the Sinomenine and acupuncture groups. The expression levels of NGF in high dose and mid dose group of Sinomenine and in acupuncture group were significantly decreased compared with that in model group, the alteration of expression levels of NGF and ICC-like cells had positive correlation
Compared with the model group, the level of PGE2 and IL-1βof prostate glandular tissues reduced in both Sinomenine and acupuncture groups, whereasβ-EP increased obviously.
Conclusion:Using the way of autoimmune reaction to induce the CP animal model, the change in pathology of the animal's prostate tissue was most similar to that of patients, and pathogenesis is relatively consistent with patients. It was a satisfactory way to induce CP animal model.
The CP animal model was present with bladder dysfunction of DI and alteration of ICC like cells in detrusor. The alteration of ICC-like cells in number and distribution was associated with the voiding dysfunction of CP rat model.
Sinomenine and acupuncture at Zhonglushu and Huiyang point can remarkably lower down the levels of PGE2 and IL-1β, increase theβ-EP level in rats with nonbacterial chronic prostatitis, reduce or eliminat inflammation of rat prostate tissue, and improve its organizational structure damage. They had an anti-fibrosis effect and play an important role in protecting prostate issue.
The results of this study were suggested that there would be interaction between ICC-like cells and NGF in detrusor, and the protective effect of Sinomenine and acupuncture on prostate tissue would influence the quantity and distribution of bladder ICC-like cells directly or by means of immunosuppression and regulation of neurotransmitters.
引文
1. Hanani M,Freund HR.Interstitial cells of Cajal--their role in pacing and signal transmission in the digestive system.Acta Physiol Scand,2000 Mov;170(3): 177-90.
2. Komuro T,Tokui K,Zhou DS.Identification of the interstitial cells of Cajal.Histol Histopathol,1996 Jul;11(3):769-86.
3. Burns AJ, Herbert TM,Ward SM,Sanders KM.Interstitial cells of Cajal in the guinea-pig gastrointestinal tract as revealed by c-Kit immunohistochemistry.Cell Tissue Res,1997 Oct;290(1):11-20.
4. Zhou DS, Komuro T.The cellular network of interstitial cells associated with the deep muscular plexus of the guinea pig small intestine. Anat Embryol,1992 Dec;186 (6):519-27.
5. Zhou DS, Komuro T.Ultrastructure of the zinc iodide-osmic acid stained cells in guinea pig small intestine.J Anat,1995 Oct;187(Pt 2):481-5.
6. Min KW,Sook S. Intestitial cells of Cajal in the human small intestine: immunochemical and ultrastructural study.Ultrastruct Pathol,2003 Mar-Apr;27(2):67-78.
7.刘奔,李兰英,庞智玲.干细胞因子和受体在神经系统中的表达及其生物学效应.生理科学进展.2001,32(2)：143-5.
8. Rich A, Miller SM, GibDIns SJ, Malysz J, Szurszewski JH, Farrugia G.Local presentation of Steel factor increases expression of c-kit immunoreactive interstitial cells of Cajal in culture. Am J Physiol Gastrointest Liver Physiol, 2003 Feb; 284(2):G313-20.
9. Maeda H,Yamagata A,Nishikawa S,Yoshinaga K,Kobayashi S,Nishi K,Nishikawa S. Requirement of c-kit for development of intestinal pacemaker system. Development,1992 Oct;116(2):369-75.
10. Kluppel M,Huizinga JD,Malysz J,Bernstein A. Developmental origin an Kit-dependent development of the interstitial cells of cajal in the mammalian small intestine. Dev Dyn,1998 Jan;211(1):60-71.
11. Komuro T,Zhou DS.Anti-c-kit protein immunoreactive cells corresponding to the interstitial cells of Cajal in the guinea-pig small intestine.J Auton Nerv Syst,1996 Nov 6;61(2):69-174.
12.Torihashi S,Nishi K,Tokutomi Y,Nishi T,Ward S,Sanders KM.Blockade of kit signaling induces transdifferentiation of interstitial cells of cajal to a smooth muscle phenotype.Gastroenterology,1999 Jul; 117(1):140-8.
13. Vannucchi MG.Receptors in interstitial cells of Cajal:identification and possible physiological roles.Microsc Res Tech,1999 Dec 1;47(5):325-35.
14.Midrio P,Vannucchi MG, Pieri L, Alaggio R, Faussone-Pellegrini MS.Delayed development of Interstitial cells of Cajal in the ileum of a human case of gastroschisis.J Cell Mol Med,2008, Apr;12(2):471-8. Epub 2008 Feb 8.
15. McCloskey KD, Gurney AM. Kit positive cells in the guinea pig bladder. J Urol, 2002 Aug;168(2):832-6.
16.Pezzone MA, Watkins SC, Alber SM, King WE, de Groat WC, Chancellor MB, Fraser MO.. Identification of c-kit-positive cells in the mouse ureter:the interstitial cells of Cajal of the urinary tract. Am J Physiol Renal Physiol,2003 May;284(5):F925-9. Epub 2003 Jan 21.
17. Metzger R, Schuster T, Till H, Franke FE, Dietz HG. Cajal-like cells in the upper urinary tract:comparative study in various species. Pediatr Surg Int,2005 Mar;21(3):169-74. Epub 2005 Jan 15.
18. Metzger R, Neugebauer A, Rolle U, Bohlig L, Till H. C-Kit receptor (CD117) in the porcine urinary tract. Pediatr Surg Int,2008 Jan; 24(1):67-76.
19. van der AA F, Roskams T, Blyweert W, Ost D, Bogaert G, De Ridder D. Identification of kit positive cells in the human urinary tract. J Urol,2004 Jun;171(6 Pt 1):2492-6.
20. Solari V, Piotrowska AP, Puri P. Altered expression of interstitial cells of Cajal in congenital ureteropelvic junction obstruction. J Urol,2003 Dec;170 (6 Pt 1): 2420-2.
21. Metzger R, Schuster T, Till H, Stehr M, Franke FE, Dietz HG. Cajal-like cells in the human upper urinary tract. J Urol,2004 Aug;172(2):769-72.
22. Blyweert W, van der Aa F, Ost D, Stagnaro M, Ridder D. Interstitial cells of the bladder:the missing link? BJOG,2004 Dec; 111(s 1):57-60.
23. David SG, Cebrian C, Vaughan ED, Herzlinger D. C-kit and ureteral peristalsis. J Urol,2005 Jan;173(1):292-5.
24. Lang RJ, Klemm MF. Interstitial cell of Cajal-like cells in the upper urinary tract. J Cell Mol Med,2005 Jul-Sep;9(3):543-56.
25. Lang RJ, Takano H, Davidson ME, Suzuki H, Klemm MF. Characterization of the spontaneous electrical and contractile activity of smooth muscle cells in the rat upper urinary tract. J Urol,2001 Jul; 166(1):329-34.
26. Sui GP, Rothery S, Dupont E, Fry CH, Severs NJ. Gap junctions and connexin expression in human suburothelial interstitial cells. BJU Int.2002 Jul; 90(1): 118-29.
27. Wiseman OJ, Fowler CJ,Landon DN. The role of the human bladder lamina propria myofibroblast. BJU Int,2003 Jan;91(1):89-93.
28. Drake MJ, Hedlund P, Andersson KE, Brading AF, Hussain I, Fowler C, Landon DN. Morphology phenotype and ultrastructure of fibroblastic cells from normal and neuropathic human detrusor:Absence of myofibroblast characteristics, J Urol. 2003 Apr; 169(4):1573-6.
29. Davidson RA, McClosky KD. Morphology and localization of interstitial cells in the guinea pig-bladder:structural relationships with smooth muscle and neurons.J Urol,2005 Apr;173(4):1385-90.
30. Shafik A, El-Sibai O, Shafik AA, Shafik I. Identification of interstitial cells of Cajal in human urinary bladder:concept of vesical pacemaker. Urology,2004 Oct; 64 (4):809-13.
31.Piaseczna Piotrowska A,Rolle U,Solari V,Puri P.Interstitial cells of Cajal in the human normal urinary bladder and in the bladder of patients with megacystis-microcolon intestinal hypoperistalsis syndrome. BJU Int,2004 Jul; 94 (1):143-6.
32.鄢俊安,卢根生,李龙坤,宋波.豚鼠膀胱组织ICCs细胞间隙连接蛋白Connexin43表达意义.第三军医大学学报,2008,30(7)：561-3.
33.沈文浩,熊恩庆,宋波,李为兵,卢根生,李龙坤,金锡御.成年豚鼠膀胱ICCs细胞起搏电流的鉴定.第三军医大学学报,2008,30(7)：567-9.
34. Exintaris B, Klemm MF, Lang RJ. Spontaneous slow wave and contractile activity of the guinea pig prostate. J Urol,2002Jul;168 (1):315-22.
35. Van der Aa F, Roskams T, Blyweert W, De Ridder D. Interstitial cells in the human prostate:A new therapeutic target? Prostate,2003 Sep 1;56(4):250-5.
36. Schwentner C, Oswald J, Lunacek A, Schlenck B, Berger AP, Deibl M, Fritsch H, Bartsch G, Radmayr C. Structural changes of the intravesical ureter in children with vesicoureteral reflux-does ischemia have a role? J Urol,2006 Nov; 176(5): 2212-8.
37.Roshani H, Dabhoiwala NF, Dijkhuis T, Lamers WH. Intraluminal pressure changes in vivo in the middle and distal pig ureter during propagation of a peristaltic wave. Urology,2002 Feb;59(2):298-302.
38. Oswald J, Schwentner C, Brenner E, Deibl M, Fritsch H, Bartsch G, Radmayr C. Extracellular matrix degradation and reduced nerve supply in refluxing ureteral endings. J Urol,2004 Sep;172(3):1099-102.
39. Schwentner C, Oswald J, Lunacek A, Fritsch H, Deibl M, Bartsch G, Radmayr C. Loss of interstitial cells of Cajal and gap junction protein connexin 43 at the vesicoureteral junction in children with vesicoureteral reflux. J Urol,2005 Nov; 174(5):1981-6.
40. Arena S, Fazzari C, Arena F, Scuderi MG, Romeo C, Nicotina PA, Di Benedetto V. Altered'active'antireflux mechanism in primary vesico-ureteric reflux:a morphological and manometric study. BJU Int,2007 Aug;100(2):407-12. Epub 2007 May 26.
41. Hashitani H, Yanai Y, Suzuki H. Role of interstitial cells and gap junctions in the transmission of spontaneous Ca2+signals in detrusor smooth muscles of the guinea-pig urinary bladder. J Physiol,2004 Sep 1;559(Pt 2):567-81.
42. Biers SM, Reynard JM, Doore T, Brading AF. The functional effects of a c-kit tyrosine inhibitor on guinea-pig and human detrusor. BJU Int,2006 Mar; 97(3):612-6.
43. Shafik A, Shafik I, el-Sibai O. Identification of c-kit-positive cells in the human prostate:the interstitial cells of Cajal. Arch Androl,2005 Sep-Oct;51(5):345-51.
44. Sergeant GP, Thornbury KD, McHale NG, Hollywood MA. Interstitial cells of Cajal in the urethra. J Cell Mol Med,2006 Apr-Jun;10(2):280-91.
1.程伟,翟君,赵养宏.慢性前列腺炎患者的尿动力学检查(附32例报告).临床泌尿外科杂志,2001,16(5)：200-1.
2.马汉彬,陈复纯,肖广潮.慢性前列腺炎患者的尿动力学检查.河北医学,2006,12(3)：205-7.
3.于洪波,孙则禹,卫中庆.尿动力学异常与慢性前列腺炎诊治关系的探讨.中华男科学2001,7(4)：240-2.
4. Chapple CR, Smith D. The pathophysiological changes in the bladder obstructed by benign prostatic hyperplasia. Br J Urol,1994 Feb;73(2):117—23.
5. Brading AF, Turner WH. The unstable bladder:towards a common mechanism. Br J Urol,1994 Jan;73(1):3—8.
6.宋波.逼尿肌兴奋性及逼尿肌不稳定.解放军医学杂志,2003,28(2)：128-30.
7.封建立,李龙坤,杨景,方强,丁砺蠡,宋波.不稳定逼尿肌中ICCs细胞与逼尿肌收缩的关系.第三军医大学学报,2008；10(30)：894-6.
8.莫志贤,贺慧洁,朱争华.青藤碱组胺释放作用与抗组胺收缩肠管作用研究.中药药理与临床,2006,22(6)：16-8.
9.陈跃来,申鹏飞,陈国美,哈力甫,宗蕾,吴晨力.电针对无菌性前列腺炎大鼠膀胱及前列腺组织病理学的影响.上海针灸杂志,2003,22(7)：6-9.
10. Theodorou C, Konidaris D, Moutzouris G, Becopoulos T.The urodynamic profile of Prostatodynia.BJU Int.1999 Sep;84(4):461-3.
11.夏同礼主编.现代泌尿病理学.北京：人民卫生出版社,第1版.2002,481-4.
12. Potts JM. Diagnosing the prostatitis patient:the dilemma continues. Curr Urol Rep,2002 Aug; 3(4):319-23.
13.周晓辉,韩蕾,周智恒,刘忠德,杨吉相,吕延伟,尤春来.免疫性慢性非细菌性前列腺炎大鼠模型的形态学与分子生物学特性.中华男科学杂志,2005,11(4)：290-5.
14.张亚强,王炎,李敏,卢建新,高筱松,刘兵,庞然.大鼠前列腺组织抗原蛋白诱导自身免疫性前列腺炎模型的建立.中国中西医结合外科杂志,2008,14(6)：584-7.
15.Keeteh DW, Humphrey P, Ratliff TL. Development of a mouse model for Nonbacterial prostatitis.J Urol,1994 Jul; 152(1):247-50.
16.Maeeioni M, Riera CM. Prostatein(or rat prostatic steroid binding protein) is a major autoantigen in experimental autoimmune prostatitis. Clin Exp Immunol, 1998 May;112(2):159-65.
17.周晓辉,周智恒,刘忠德.自身免疫因素导致慢性非细菌性前列腺炎的研究进展.中国中西医结合外科杂志,2003,9(3)：240-2.
18.叶伟成,薛慈民,徐兆东,王耀萍,肖志灏,付胜光.免疫佐剂法制作慢性非细菌性前列腺炎小鼠模型的方法.中国男科学杂志,2001,15(1)：29-32.
19. Andersson KE. Pharmacology of lower urinary tract smooth muscles and penile erectile tissues. Pharmacol Rev,1993 Sep;45(3):253-308.
20. Brading AF. Ion channels and control of contractile activity in urinary bladder smooth muscle. Jpn J Pharmacol,1992;58 Suppl 2:120-7.
21. Drake MJ, Harvey IJ, Gillespie JI. Autonomous activity in the isolated guinea pig bladder. Exp Physiol,2003 Jan;88(1):19-30.
22. Drake MJ, Mills IW, Gillespie JI. Model of peripheral autonomous modules and a myovesical plexus in normal and overactive bladder function. Lancet,2001 Aug 4;358(9279):401-3.
23.Herrera GM, Heppner TJ, Nelson MT. Regulation of urinary bladder smooth muscle contractions by ryanodine receptors and BK and SK channels. Am J Physiol Regu Integr Comp Physiol,2000 Jul;279(1):R60-8.
24.Imai T, Okamoto T, Yamamoto Y, Tanaka H, Koike K, Shigenobu K, Tanaka Y. Effects of different types of K+ channel modulators on the spontaneous myogenic contraction of guineapig urinary bladder smooth muscle.Acta Physiol Scand,2001 Nov;173(3):323-33.
25. Imai T, Tanaka Y, Okamoto T, Yamamoto Y, Horinouchi T, Tanaka H, Koike K, Shigenobu K. Evidence that action potential generation is not the exclusive determinan to trigger spontaneous myogenic contraction of guinea-pig urinary bladder smooth muscle. Acta Physiol Scand,2002 Sep;176(1):57-63.
26. Hanani M, Freund HR. Interstitial cells of Cajal—their role in pacing and signal transmission in the digestive system.Acta Physiol Scand,2000 Nov;170(3): 177-90.
27. McCloskey KD, Alison GM. Kit positive cells in the guinea pig bladder. J Urol, 2002 Aug;168(2):832-36.
28. Pezzone MA, Watkins SC, Alber SM, King WE, de Groat WC, Chancellor MB, Fraser MO. Identification of c-kit-positive cells in the mouse ureter:the interstitial cells of Cajal of the urinary tract. Am J Physiol,2003 May;284(5): F925-9. Epub 2003 Jan 21.
29. Metzger R, Schuster T, Till H, Franke FE, Dietz HG. Cajal-like cells in the upper urinary tract:comparative study in various species.Pediatr Surg Int,2005 Mar;21(3):169-74. Epub 2005 Jan 15.
30. Van Der AA F, Roskams T, Blyweert W, Ost D, Bogaert G, De Ridder D Identification of kit positive cells in the human urinary tract.J Urol,2004 Jun;171(6 Pt 1):2492-6.
31. Sergeant G P, Hollywood M A, McCloskey K D, Thornbury KD, McHale NG. Specialised pacemaking cells in the rabbit urethra.J Physiol,2000 Jul 15;526(Pt 2):359-66.
32. Kubota Y, Biers SM, Kohri K, Brading AF. Effects of imatinib mesylate (Glivec) as a c-kit tyrosine kinase inhibitor in the guinea-pig urinary bladder. Neurourol Urodyn,2006; 25 (3):205-10.
33. Biers SM, Reynard J M, Doore T, Brading AF. The functional effects of a c-kit tyrosine inhibitor on guinea-pig and human detrusor. BJU Int,2006 Mar;97 (3) 612-6.
34.路扬,李龙坤,王永权.牵张负荷对大鼠膀胱ICCs细胞数量及c-kit表达的影响.第三军医大学学报,2008,30(7)：570-3
35.丁砺蠡,方强,封建立,宋波.膀胱Cajal间质细胞对膀胱逼尿肌细胞兴奋调控的结构和功能特征的初步研究.第三军医大学学报,2008,30(12)：1118-21.
36.鄢俊安,卢根生,李龙坤,宋波.豚鼠膀胱组织ICCs细胞间隙连接蛋白Connexin 43表达意义.第三军医大学学报,2008,30(7)：561-3.
1 Nadler RB, Koch AE, Calhoun EA, Campbell PL, Pruden DL, Bennett CL, Yarnold PR, Schaeffer AJ. IL-lbeta and TNF-alpha in prostatic secretions are indicators in the evaluation of men with chronic prostatitis. J Urol,2000 Jul; 164 (1):214-8.
2 Shahed AR, Shoskes DA. Correlation of beta-endorphin and prostaglandin E2 levels in prostatic fluid of patients with chronic prostatitis with diagnosis and treatment response. J Urol,2001 Nov;166(5):1738-41.
3 王翔宇,王佩芳,金亚萍,施秀丽,蔡定芳.会阳、中膂俞在下尿路功能性疾病中的临床应用.针灸临床杂志,2004,20(11)41-2.
4 石德光,杜欣,胡森.针刺对慢性炎症疾病中炎症介质的调控及机制.中国临床康复,2002,6(20)：3040-1.
5 冀来喜.针刺秩边穴对试验性非细菌性前列腺炎大鼠前列腺微循环的影响.中国针灸,2001；21(1)：45-6.
6 张宝文,周景华,沈梅,姜松鹤,王建明,李成永,陈晓云.取热刺法对大鼠实验性前列腺炎组织学及超微结构的影响.中国中医药科技,2004,11(6)：364-5.
7 陈跃来,申鹏飞,陈国美,哈力甫,宗蕾,吴晨力.电针对无菌性前列腺炎大鼠膀胱及前列腺组织病理学的影响.上海针灸杂志,2003,22(7)：6-9.
8 张宝文,王迎新,沈梅,姜松鹤,王建明,高滨昌,李成永,陈晓云.取热刺法对大鼠实验性前列腺炎白细胞介素-2及血清睾酮含量的影响.中国中医药科技,2004,11(6)：365-36.
9 林文注,王佩.实验针灸学.第1版.上海：上海科技出版社,1999：284-8,183-7.
10 李锦宇,韩霞,汪晓斌.针灸免疫研究概况.动物医学进展,2008,29(11)：107-10.
11 Li Y, Cui S, Cheng Y, Chen X, Hu Z. Application of nonaqueous capillary electrophoresis for quantitative analysis of quinolizidine alkaloids in Chinese herbs. Anal Chim Acta,2004 Apr;508(1):17-22.
12 Liu L, Buchner E, Beitze D, Schmidt-Weber CB, Kaever V, Emmrich F, Kinne RW. Amelioration of rat experimental arthritides by treatment with the alkaloid sinomenine. Int J Immunopharmacol,1996 Oct;18(10):529-43.
13 Zhao Y, Li J, Yu K, Liu Y, Chen X. Sinomenine inhibits maturation of monocyte-derived dendritic cells through blocking activation of NF-kappa B. Int Immunopharmacol,2007 May;7(5):637-45. Epub 2007 Feb 12.
14 Shu L, Yin W, Zhang J, Tang B, Kang YX, Ding F, Hua ZC. Sinomenine inhibits primary CD4+T-cell proliferation via apoptosis. Cell Biol Int,2007 Aug; 31(8): 784-9. Epub 2007 Jan 30.
15 Feng H, Yamaki K, Takano H, Inoue K, Yanagisawa R, Yoshino S. Suppression of Thl and Th2 immune responses in mice by Sinomenine, an alkaloid extracted from the chinese medicinal plant Sinomenium acutum. Planta Med,2006 Dec; 72(15):1383-8. Epub 2006 Nov 7.
16 Liu L, Riese J, Resch K, Kaever V. Impairment of macrophage eicosanoids and nitric oxide production by alkaloid from Sinomenium acutum. Arzneimittel-forschung,1994 Nov;44(11):1223-6.
17 李晓娟,王培训,刘良,陈光星,肇静娴,曾耀英,陈纪藩.青藤碱对T淋巴细胞活化及TH1类细胞内细胞因子表达的影响.中国免疫学杂志,2004,20(4)：249-58.
18 王毅,陈正,熊烈,罗志刚,秦国庆,李建军.青藤碱对肾移植大鼠急性排斥反应及T细胞增殖的影响.中华实验外科杂志,2004,21(5)：573-4.
19 He X, Wang J, Guo Z, Liu Q, Chen T, Wang X, Cao X. Requirement for ERK activation in sinomenine-induced apoptosis of macrophages. Immunol Lett,2005 Apr 15;98(1):91-6. Epub 2004 Nov 25.
20 徐道华,周晨慧.树突状细胞CD80和CD86表达及胞外白细胞介素12分泌与不同剂量青藤碱干预的影响.中国组织工程研究与临床康复,2007,11(28)：5654-6.
21 杨承英,陈永文,傅晓岚,费蕾,靳斯,谢谆怡,汤玉渝,吴玉章.青藤碱促进树突状细胞分化抑制其成熟.免疫学杂志,2007,23(3)：265-9.
22 赵毅,余克强,李娟.青藤碱对类风湿关节炎树突状细胞核转录因子-κB活性的影响.广东医学,2006,27(1)：55-7。
23 高永翔,龚立,田艳勋,杨雪,汤洪萍.青藤碱对小鼠骨髓树突状细胞免疫功能的影响.成都中医药大学学报,2005,28(3)：13-4.
24 梁瑞燕,曹柳英,王文君,刘新迎,王培训.青藤碱抗炎作用机理研究.广州中医药大学学报,2007,24(2)：141-3.
25 王文君,王培训.青藤碱对环氧化酶2活性的选择性抑制作用.广州中医药大学学报,2002,19(1)：46-7.
26 陈炜,沈悦娣,赵光树,姚杭平.青藤碱对脂多糖诱导的神经细胞环氧化酶-2表达的影响.中国中药杂志,2004,29(9)：900-3.
27 涂胜豪,胡永红,陆付耳.青藤碱对人淋巴细胞产生IL-2,IL-2R和IL-6的影响. 中国实验临床免疫学杂志,1998,10(5)：268-70.
28 刘良,李晓娟,王培训,王文君,周联,梁瑞燕,曹柳英.青藤碱对人外周血单个核细胞IL-1β和IL-8两细胞因子基因表达的影响.中国免疫学杂志,2002,18(4)：241-4.
29 Lang MD, Nickel JC, Olson ME, Howard SR, Ceri H. Rat model of experimentally induced abacterial prostatitis. Prostate,2000 Nov 1;45(3):201-6.
30 Vykhovanets EV, Shukla S, MacLennan GT, Vykhovanets OV, Bodner DR, Gupta S. IL-1 beta-induced post-transition effect of NF-kappaB provides time-dependent wave of signals for initial phase of intrapostatic inflam-mation. Prostate,2009 May 1;69(6):633-43.
31 陈曦,李凡.Glucosamine Chondroitin与黄芩联合对佐剂性关节炎大鼠血清PGE2影响的实验研究.中国实验诊断学,2005,2(9)：15-7.
32 Takagi-Matsumoto H, Ng B, Tsukimi Y, Tajimi M. Effects of NSAIDs on bladder function in normal and cystitis rats:a comparison study of aspirin, indomethacin, and ketoprofen. J Pharmacol Sci,2004 Aug; 95(4):458-65.
33 Velasco C, Angelico P,Guarneri, L, Leonardi A, Clarke DE, Testa R. Effects of the nuclear factor-kappaB inhibitors 2-hydroxy-4-trifluoromethylbenzoic acid and aspirin on micturition in rats with normal and inflamed bladder. J Urol, 2001,166:1962-68.
34 王勒,欧可群.内阿片肽的研究进展.四川解剖学杂志,2000,4：227-30.
1.金锡御,宋波.临床尿动力学.第1版.北京：人民卫生出版社,2001.165-175.
2. Brdaing AF, Tunrer WH. The unstable bladder:towards a common mechanism. Br J Uorl,1994 Jan;73(1):3-8.
3. Harrison SCW, Hunnam GR, Farman P, Ferguson DR, Doyle PT. Bladder instability and denervation in patiens with bladder outflow obstruction.Br J Urol, 1987 Dec;60(6):519-22.
4. Emmons SL, Otto L. Acupuncture for overactive bladder:a randomized controlled trial.Obstet Gynecol.2005 Jul; 106(1):138-43.
5.陈跃来,宗蕾,陈国美,申鹏飞.针刺对28例不稳定膀胱尿流动力学影响的观察.浙江中医杂志,2002,37(11)：490-1.
6.陈跃来,岑珏,侯文光,高志强,虞先敏,马雪梅.电针对大鼠不稳定膀胱逼尿肌及膀胱颈氮能神经递质的影响.中西医结合学报,2006,4(1)：73-5.
7.王翔宇,郑蕙田,汪司右,唐燕红,李建忠,施秀丽.电针中膂俞抑制膀胱活动亢进的传入神经机制.上海针灸杂志,2003,22(1)：17-20.
8. Nishida S, Satoh H. Vascular pharmacology of mokuboito (mu-fang-yi-tang) and its constituents on the smooth muscle and the endothelium in rat aorta. Evid Based Complement Alternat Med.2007 Sep;4(3):335-41.
9. Wang CY, Mo ZX, Tu HH. Effect of sinomenine on morphine dependence in isolated guinea pig ileum. Di Yi Jun Yi Da Xue Xue Bao.2003 Apr; 23(4):329-31.
10. de Groat WC.Central neural control of the lower urinary tract. Ciba Found Symp. 1990;151:27-44; discussion 44-56.
11.Brading A F. A myogenic basis for the overactive bladder. Urology,1997 Dec;50(6A Suppl):57-67; discussion 68-73.
12. McCloskey KD, GurneyAM. Kit positive cells in the guinea pig bladder. J Urol, 2002 Aug;168 (2):832-6.
13. Metzger R, Schuster T, Till H, Stehr M, Franke FE, Dietz HG. Cajal-like cells in the human upper urinary tract. J Urol,2004 Aug;172 (2):769-72.
14. Popescu LM, Gherghiceanu M, Cretoiu D, Radu E. The connective connection: interstitial cells of Cajal (ICC) and ICC-like cells establish synapses with immunoreactive cells. Electron microscope study in situ. J Cell Mol Med,2005 Jul-Sep;9 (3):714-30.
15. Davidson RA, McCloskey KD. Morphology and localization of interstitial cells in the guinea pig bladder:Structural relationships with smooth muscle and neurons.J Urology,2005 Apr;173(4):1385-90.
16.Min KW, Sook S. Intestitial cells of Cajal in the human small intestine: immunochemical and ultrastructural study. Ultrastruct Pathol,2003 Mar-Apr;27(2):67-78.
17. McCloskey KD. Interstitial cells in the urinary bladder--localization and function. Neurourol Urodyn.2010;29(1):82-7.
18. Van Der AA F, Roskams T, Blyweert W, Ost D, Bogaert G, De Ridder D. Identification of kit positive cells in the human urinary tract.J Urol,2004 Jun;171(6 Pt 1):2492-6.
1.张海明,张映.神经生长因子对神经元作用的研究进展动物医学进展,2006,27(9)：39-41.
2. Lamb K, Gebhart GF, Bielefeldt K. Increased nervegrowth factor expression triggers bladder overactivity. J Pain,2004 Apr;5 (3):150—6.
3. Potts JM. Diagnosing the prostatitis patient:the dilemma continues. Curr Urol Rep.2002 Aug; 3 (4):319-23.
4. Brdaing AF, Tunrer WH. The unstable bladder:towards a common mechanism. Br J Uorl,1994 Jun;73(1):3-8.
5. Harrison SCW, Hunnam GR, Farman P, Ferguson DR, Doyle PT. Bladder instability and denervation in patiens with bladder outflow obstruction.Br J Urol, 1987 Dec;60(6):519-22.
6. Dvaies AM. Nerve growth factor synthesis and nerve growth factor receptor expression in neural development.Int Rev Cytol 1991; 128:109-38.
7. Theodorou V, Fioramonti J, Bueno L.Integrative neuroimmunology of the digestive tract.Vet Res.1996;27(4-5):427-42.
8. Creedon D, Tuttle JB. Nerve growth factor synthesis in vascular smooth musclel. Hypertension.1991; 18(6):730-41.
9. Spitsbergen JM, Stewart JS, Tuttle JB. Altered regulation of nerve growth factor secretion by cultured VSMCs from hypertensive rats. Am J Physiol.1995 Aug;269(2 Pt 2):H621-8.
10. Liu HT, Chancellor MB, Kuo HC. Urinary nerve growth factor levels are elevated in patients with detrusor overactivity and decreased in responders to detrusor botulinum toxin-A injection. Eur Urol.2009 Oct;56(4):700-6.
11.刘孝东,何育霖,申吉泓,王黎,张林,张建华.神经生长因子在脊髓高位损伤大鼠膀胱中表达量的变化与意义.昆明医学院学报,2009, (10)：37-41.
12. Gosling JA, Gilpin SA, Dixon JS, Gilpin CJ. Decrease in the autonomic innervation of Human detrusor muscle in the outflow obstruction. J Urol.1986 Aug;136(2):501-4.
13.刘犇,郑宝钟,周尊林,徐忠华蔡松良.前列腺增生所致膀胱出口梗阻病人逼尿肌中NGF-mNRA的变化及意义.中华外科杂志,2004；42(14)：874-6.
14. Tanner R, Chambers P, Khadra MH, Gillespie JI. The production of nerve growth factor by human bladder smooth muscle cells in vivo and in vitro. Br J Urol Int 2000 Jun;85(9):1115-9.
15.Naoki Yoshimura, Nelson E. Bennett,Yukio Hayashi, Teruyuki Ogawa, Osamu Nishizawa, Michael B. Chancellor, William C. de Groat, Satoshi Seki. Bladder overactivity and hyperexcitability of bladder afferent neurons after intrathecal delivery of nerve growth factor in rats. J. Neurosci.2006 Oct; 26(42):10847-55.
16.Metzger R, Schuster T, Till H, Stehr M, Franke FE, Dietz HG. Cajal-like cells in the human upper urinary tract. J Urol,2004 Aug;172 (2):769-72.
17. Davidson RA, McCloskey KD. Morphology and localization of interstitial cells in the guinea pig bladder:Structural relationships with smooth muscle and neurons.J of Urology,2005 Apr;173:1385-90.
18.Min KW, Sook S. Intestitial cells of Cajal in the human small intestine: immunochemical and ultrastructural study. Ultrastruct Pathol,2003 Mar-Apr;27(2):67-78.
19. McCloskey KD. Interstitial cells in the urinary bladder--localization and function. Neurourol Urodyn.2010;29(1):82-7.
20. Popescu LM, Gherghiceanu M, Cretoiu D, Radu E. The connective connection: interstitial cells of Cajal (ICC) and ICC-like cells establish synapses with immunoreactive cells. Electron microscope study in situ. J Cell Mol Med,2005 Jul-Sep;9 (3):714-30.