缩泉丸对自然衰老大鼠膀胱逼尿肌受体的调控及机制研究
摘要
在临床上,膀胱逼尿肌功能障碍引起的尿频、尿急、尿失禁和遗尿极为常见。而逼尿肌收缩性、兴奋性和顺应性是影响膀胱功能的主要因素。缩泉丸出自《妇人良方》,由乌药、益智仁、山药组成。主治尿多,尿频,遗尿。具有温化肾气,缩尿止遗的功效,其功效可概括为“补肾缩尿”。研究已经证实,直接介导膀胱逼尿肌收缩的主要受体是M3R,介导膀胱逼尿肌松弛的主要受体是β3-AR。
目的：以调节膀胱逼尿肌细胞收缩功能的M3R及其介导的信号转导通路和调节膀胱逼尿肌细胞舒张功能的p3-AR及其介导的信号转导通路为出发点,立足于逼尿肌功能异常与神经源性的相关性,以膀胱逼尿肌细胞为研究对象,阐明缩泉丸“缩尿”功效与现代医学调节逼尿肌舒缩功能的内在联系。
方法：
以自然衰老大鼠为“肾虚多尿”模型。取筛选合格的自然衰老大鼠(15月龄)70只,青年大鼠(3月龄)14只。设置组别：青年大鼠为正常对照组,14只,自然衰老大鼠分为模型对照组、金匮肾气丸组、缩泉丸低、中、高剂量组,14只/组。金匮肾气丸组的给药剂量为1.08g/kg/d;缩泉丸低、中、高剂量分别为0.293g/kg/d、0.585g/kg/d、1.170g/kg/d。采用酶消化与组织块相结合的方法原代培养自然衰老大鼠膀胱逼尿肌细胞；采用RT-PCR方法测定膀胱逼尿肌细胞M3R和β3-AR mRNA的表达；采用western blotting方法测定膀胱逼尿肌细胞M3R和β3-AR蛋白的表达；采用酶联免疫分析试剂盒测定膀胱逼尿肌细胞IP3含量；采用激光扫描共聚焦显微镜测定膀胱逼尿肌细胞Ca2+和CaM浓度。
结果：
1、缩泉丸减少自然衰老大鼠的尿量,提示缩泉丸有“缩尿”的作用；
2、成功地原代培养出自然衰老大鼠膀胱逼尿肌细胞；
3、缩泉丸能使自然衰老大鼠逼尿肌细胞M3R mRNA表达降低,β3-AR mRNA表达增加；
4、缩泉丸能使自然衰老大鼠逼尿肌细胞M3R蛋白表达降低,β3-AR蛋白表达增加；
5、缩泉丸能使自然衰老大鼠逼尿肌细胞的IP3的含量降低；
6、缩泉丸使H-89阻断β3-AR介导的信号通路的自然衰老大鼠逼尿肌细胞PKA蛋白表达增加；
7、缩泉丸使自然衰老大鼠逼尿肌细胞Ca2+和CaM浓度均降低。
结论：
缩泉丸下调M3R及其介导的信号转导通路和上调β3-AR及其介导的信号转导通路,从而维持了二者的动态平衡,缩泉丸能使自然衰老大鼠逼尿肌细胞Ca22+和CaM浓度均下调,降低逼尿肌的收缩力和兴奋性,减少逼尿肌异常收缩频率,保证了膀胱正常的生理功能。将缩泉丸“缩尿”理论结合神经源性来探讨缩泉丸作用的多靶性,为缩泉丸治疗肾虚多尿提供了确切的分子药理学基础,为中医“补肾缩尿”的理论给予新的诠释。
Frequent urination,urgency of urination,urinary incontinence and enuresis caused by Bladder detrusor dysfunction are very common in the clinic. The main factor affecting the bladder function is contractility, excitability and compliance of detrusor.SQW is reeorded in Chinese medicine book named Furenliangfang. SQW consists of three herb, Radix Linderae, Radix dioscoreae and Alinia oxyphylla Miqfrom.The function of SQW is polyuria,urinary frequency and enuresis,and is to warm kidney,stop Polyurial and hold exeessive urination.Its effect can be summarized as" tonifying kidney and shrinkaging urinary". Studies have confirmed that the bladder detrusor contraction is directly mediated by M3muscarinic acetylcholine receptor (M3R) and the detrusor relaxation is directly mediated by β3-adrenoceptor(β3-AR).
Objective:This study was intended to regard β3-AR regulating bladder detrusor relaxation and its signal transduction pathway and regard M3R regulating bladder detrusor contraction and its signal transduction pathway as cut-in point of the research. This study was based on the dependablity of dysfunction and neurogenic of the bladder detrusor and the bladder detrusor cells was regarded as the research object. The research was elucidated the immanent connection of" reducing urination " effect and regulating diastolic and systolic function on modern medical.
Methods:Natural aging rats were used as " kidney deficiency and urinary " model. After preliminary screening,70natural aging rats were15months old,14young rats were3months old. The Setting groups were as follows:young rats were regarded as normal control group and natural aging rats were divided into model control group, Jinkui Sheni Pill,SQW low dose group, SQW middle dose group and SQW high dose group on average. The dosage of Jinkui Sheni Pill was1.08g/kg/d. The dosage of Suoquan Wan low, middle and high dose group was respectively0.293g/kg/d,0.585g/kg/d,1.170g/kg/d. primary culture of natural senescent rats detrusor cell was cultured by the combination method of enzymatic digestion and tissue. M3R and β3-AR mRNA expression of urinary bladder detrusor cells were determined by RT-PCR.M3R and P3-AR protein expression of urinary bladder detrusor cells were determined by Western blotting. IP3content of urinary bladder detrusor cells was determined by enzyme-linked immunoassay reagent kit. Ca-and CaM concentration of urinary bladder detrusor cells were determined by confocal laser scanning microscope
Results:
1. Suoquan Wan reduced urine volume of natural aging rat.This prompt " reducing urination " effect of SQW.
2. Bladder detrusor cells of Natural aging rat were easily cultured.
3. SQW could make M3R mRNA expression of detrusor muscle cells of the natural aging rats decreased,and β3-AR mRNA expression of detrusor muscle cells of the natural aging rats increased.
4. SQW could make M3R protein expression of detrusor muscle cells of the natural aging rats decreased,and β3-AR protein expression of detrusor muscle cells of the natural aging rats
5. SQW could make IP3content of detrusor muscle cells of the natural aging rats decreased.
6. SQW could make PKA protcin content of blocking signal pathway by H-89increased.
7. SQW could make Ca2+and CaM concentrations of detrusor muscle cells of the natural aging rats decreased.
SQW could make M3R and its mediated signal transduction pathway down-regulated,makeβ3-AR and its mediated signal transduction pathway up-regulated and make the Ca2+and CaM concentrations of rat detrusor cells of natural aging down-regulated. SQW reduced detrusor contractility and abnormalities contraction frequency, guaranteed the normal physiological function of bladder. The " reducing urination " effect was combined with neurogenic to discuss multiple target of SQW.It provided exact molecular pharmacology basis for SQW treating "kidney deficiency and urinary " and gived "tonifying kidney and shrinkaging urinary" theory to a new interpretation
目的：以调节膀胱逼尿肌细胞收缩功能的M3R及其介导的信号转导通路和调节膀胱逼尿肌细胞舒张功能的p3-AR及其介导的信号转导通路为出发点,立足于逼尿肌功能异常与神经源性的相关性,以膀胱逼尿肌细胞为研究对象,阐明缩泉丸“缩尿”功效与现代医学调节逼尿肌舒缩功能的内在联系。
方法：
以自然衰老大鼠为“肾虚多尿”模型。取筛选合格的自然衰老大鼠(15月龄)70只,青年大鼠(3月龄)14只。设置组别：青年大鼠为正常对照组,14只,自然衰老大鼠分为模型对照组、金匮肾气丸组、缩泉丸低、中、高剂量组,14只/组。金匮肾气丸组的给药剂量为1.08g/kg/d;缩泉丸低、中、高剂量分别为0.293g/kg/d、0.585g/kg/d、1.170g/kg/d。采用酶消化与组织块相结合的方法原代培养自然衰老大鼠膀胱逼尿肌细胞；采用RT-PCR方法测定膀胱逼尿肌细胞M3R和β3-AR mRNA的表达；采用western blotting方法测定膀胱逼尿肌细胞M3R和β3-AR蛋白的表达；采用酶联免疫分析试剂盒测定膀胱逼尿肌细胞IP3含量；采用激光扫描共聚焦显微镜测定膀胱逼尿肌细胞Ca2+和CaM浓度。
结果：
1、缩泉丸减少自然衰老大鼠的尿量,提示缩泉丸有“缩尿”的作用；
2、成功地原代培养出自然衰老大鼠膀胱逼尿肌细胞；
3、缩泉丸能使自然衰老大鼠逼尿肌细胞M3R mRNA表达降低,β3-AR mRNA表达增加；
4、缩泉丸能使自然衰老大鼠逼尿肌细胞M3R蛋白表达降低,β3-AR蛋白表达增加；
5、缩泉丸能使自然衰老大鼠逼尿肌细胞的IP3的含量降低；
6、缩泉丸使H-89阻断β3-AR介导的信号通路的自然衰老大鼠逼尿肌细胞PKA蛋白表达增加；
7、缩泉丸使自然衰老大鼠逼尿肌细胞Ca2+和CaM浓度均降低。
结论：
缩泉丸下调M3R及其介导的信号转导通路和上调β3-AR及其介导的信号转导通路,从而维持了二者的动态平衡,缩泉丸能使自然衰老大鼠逼尿肌细胞Ca22+和CaM浓度均下调,降低逼尿肌的收缩力和兴奋性,减少逼尿肌异常收缩频率,保证了膀胱正常的生理功能。将缩泉丸“缩尿”理论结合神经源性来探讨缩泉丸作用的多靶性,为缩泉丸治疗肾虚多尿提供了确切的分子药理学基础,为中医“补肾缩尿”的理论给予新的诠释。
Frequent urination,urgency of urination,urinary incontinence and enuresis caused by Bladder detrusor dysfunction are very common in the clinic. The main factor affecting the bladder function is contractility, excitability and compliance of detrusor.SQW is reeorded in Chinese medicine book named Furenliangfang. SQW consists of three herb, Radix Linderae, Radix dioscoreae and Alinia oxyphylla Miqfrom.The function of SQW is polyuria,urinary frequency and enuresis,and is to warm kidney,stop Polyurial and hold exeessive urination.Its effect can be summarized as" tonifying kidney and shrinkaging urinary". Studies have confirmed that the bladder detrusor contraction is directly mediated by M3muscarinic acetylcholine receptor (M3R) and the detrusor relaxation is directly mediated by β3-adrenoceptor(β3-AR).
Objective:This study was intended to regard β3-AR regulating bladder detrusor relaxation and its signal transduction pathway and regard M3R regulating bladder detrusor contraction and its signal transduction pathway as cut-in point of the research. This study was based on the dependablity of dysfunction and neurogenic of the bladder detrusor and the bladder detrusor cells was regarded as the research object. The research was elucidated the immanent connection of" reducing urination " effect and regulating diastolic and systolic function on modern medical.
Methods:Natural aging rats were used as " kidney deficiency and urinary " model. After preliminary screening,70natural aging rats were15months old,14young rats were3months old. The Setting groups were as follows:young rats were regarded as normal control group and natural aging rats were divided into model control group, Jinkui Sheni Pill,SQW low dose group, SQW middle dose group and SQW high dose group on average. The dosage of Jinkui Sheni Pill was1.08g/kg/d. The dosage of Suoquan Wan low, middle and high dose group was respectively0.293g/kg/d,0.585g/kg/d,1.170g/kg/d. primary culture of natural senescent rats detrusor cell was cultured by the combination method of enzymatic digestion and tissue. M3R and β3-AR mRNA expression of urinary bladder detrusor cells were determined by RT-PCR.M3R and P3-AR protein expression of urinary bladder detrusor cells were determined by Western blotting. IP3content of urinary bladder detrusor cells was determined by enzyme-linked immunoassay reagent kit. Ca-and CaM concentration of urinary bladder detrusor cells were determined by confocal laser scanning microscope
Results:
1. Suoquan Wan reduced urine volume of natural aging rat.This prompt " reducing urination " effect of SQW.
2. Bladder detrusor cells of Natural aging rat were easily cultured.
3. SQW could make M3R mRNA expression of detrusor muscle cells of the natural aging rats decreased,and β3-AR mRNA expression of detrusor muscle cells of the natural aging rats increased.
4. SQW could make M3R protein expression of detrusor muscle cells of the natural aging rats decreased,and β3-AR protein expression of detrusor muscle cells of the natural aging rats
5. SQW could make IP3content of detrusor muscle cells of the natural aging rats decreased.
6. SQW could make PKA protcin content of blocking signal pathway by H-89increased.
7. SQW could make Ca2+and CaM concentrations of detrusor muscle cells of the natural aging rats decreased.
SQW could make M3R and its mediated signal transduction pathway down-regulated,makeβ3-AR and its mediated signal transduction pathway up-regulated and make the Ca2+and CaM concentrations of rat detrusor cells of natural aging down-regulated. SQW reduced detrusor contractility and abnormalities contraction frequency, guaranteed the normal physiological function of bladder. The " reducing urination " effect was combined with neurogenic to discuss multiple target of SQW.It provided exact molecular pharmacology basis for SQW treating "kidney deficiency and urinary " and gived "tonifying kidney and shrinkaging urinary" theory to a new interpretation
引文
[1]李淑雯.缩泉丸对醛固酮合成与分泌的调控机制研究[D].广州中医药大学博士研究生论文,2009年
[2]谭莹.基于逼尿肌胶原蛋白及神经受体探讨缩泉丸作用机制[D].广州中医药大学博士研究生论文,2009年.
[3]操红缨.基于AQP_2研究缩泉丸补肾缩尿的机理[D].广州中医药大学博士研究生论文,2008年
[4]王米渠,胡聪.试论肾主水司二阴并及肾体[J].片肃中医,1998,11(5)：1-4.
[5]李奕祺.“精水合一”是肾藏象的认识基础浅识[J].中医药学刊,2004,22(2)：302
[6]陈明,龙子江,卞海.补肾缩泉胶囊对老年性夜尿增多症模型及老年大鼠尿量的影响[J].中国中医药科技,2007；14(5)：337-339
[7]吕圭源.药理学(第2版)[M].北京：中国中医药出版社,2007：149.
[8]朱大年.生理学(第7版)[M].北京：人民卫生出版社,2008：212-230.
[9]操红缨,吴清和,黄萍,等.缩泉丸对肾阳虚多尿大鼠尿BUN, Cr、Na+、K+、Cl离子浓度的影响[J].中医药临床杂志,2009；21(2)：117.
[10]操红缨,吴清和,黄萍,等.缩泉丸对肾阳虚多尿大鼠肾脏病理的影响[J].中国现代药物应用.2009；3(14)：59-60.
[11]刘春蓓,胡伟新.肾素-血管紧张素系统：新成员,新效应[J].肾脏病与透析肾移植杂志,2005；14(4)：349-355.
[12]吴玉付,李醒三.肾素-血管紧张素-醛固酮系统在心血管病中的作用[J].医学文选,2007；24(6)：1023-1027.
[13]刘建萍,刘精东.肾素血管紧张素系统与糖尿病肾病相关性研究进展[J].江西医药,2008；43(8)：851-854.
[14][19] Taufiq-Ur-Rahman M,Shilpi JA,Ahmed M,et al.Preliminary Pharmacological studies on Piper chaba stem bark.J Ethnopharmacol,2005;99(2):203-209.
[15]刘进平,韩涛,浦晓东.血.管紧张素Ⅱ受体研究进展[J].福建医药杂志,2006；28(4)：97-100.
[16]叶绪英,宋卉,卢成志.葛根素注射液对自发性高血压大鼠AT1和ACE2mRNA表达的影响[J].中国中西医结合杂志,2008；28(9)：824-828.
[17]李淑雯,吴清和,黄萍,等.缩泉丸对肾虚多尿大鼠肾脏MRmRNA及蛋白表达的影响[J].中药药理与临床,2010；26(6)：8-10.
[18]SoPhie Combet,Nancy Geffroy,Veronique Berthonaud.Correction of Age-related Polyuria by dDAVP:molecular analysis of aquaporins and urea transporters.Am J Physiol Renal Physiol,2003;284:F199-F208.
[19]Katsuya Suzuki,Ryo Hatano,Mari Michimata,et al.Residual Urinary Concentrat-ing Ability and AQP2 Expression in a Rat Model for Chronie Renal Failure. Neph-ron Physiol 2005;(99):P16-P22.
[20]操红缨,吴清和,黄萍,等.缩泉丸对肾阳虚多尿大鼠肾脏AQP-2与AVPR-V2mRNA表达的影响[J].中药材,2009；2(6)：926-928.
[21]连晓媛,陈奇.应激与中枢神经递质及HPA轴功能调节[J].国外医学·生理、病理科学与临床分册,1998；18(4)：371-374.
[22]李淑雯,吴清和,黄萍,等.腺嘌呤对大鼠下丘脑-垂体-肾上腺皮质(HPA)轴的影响[J].时珍国医国药,2010；21(9)：2358.
[23]操红缨,吴清和,黄萍,等.缩泉丸对肾阳虚多尿大鼠内分泌及免疫功能的影响[J].中药新药与临床药理.2009；20(4)：323-326.
[24]Jun-Ho La,Tae-Sik Sung,Hyun-Ju Kim.Peripheral corticotropin releasing Hormone mediates Post-inflammatory visceral hypersensitivity in rats. (?)世界胃肠病学杂志《英文版》,2008；14(5)：731-736.
[25]郑小伟,包素珍,李荣群,等.二仙汤对肾阳虚大鼠垂体ACTH基因表达的影响[J].中国医药学报,2003；18(12)：716-719.
[26]张秀珍.内分泌代谢疾病与肾脏[M].上海：复旦大学出版社,2004：50.
[27]赖凌云,顾勇.醛固酮的新认识[J].国外医学·生理、病理科学与临床分册.2002；22(4)：347-349.
[28]李晓东,唐德集,刘华锋,等.醛固酮与肾脏疾病[J].国外医学·生理、病理科学与临床分册,2005；25(3)：232-236.
[29]King LS,Kozono D,Agre P:From structure to disease.The evolving tale of Aquaporin biology.Nat Rev Mol Cell Biol 2004:5:687-698.
[30]Klussmann E,Maric K,Wiesner B,et al.Protein kinase A anchoring Proteins areRequired for vasoPressin-mediated translocation of aquaporin-2 into cell membranes of renal Principal cells.J BiolCheml999:274:4934-4938.
[31]S.Rttig,J.Frokier.Basis and therapeutical rationale of the urinary Concentrating mechanism.Int J Clin Pract,2007,61(Supp1·155),2-7.
[32]操红缨,吴清和,黄萍,等.缩泉九对肾阳虚多尿模型大鼠血cAMP和ALD水平的影响[J].新中医,2009；41(10)：101.
[33]于赫,郭丽新.肾阳虚证的生物化学与分子生物学研究进展[J].中医药信息,2005；22(6)：41-44.
[34]林茂樟,临床肾脏生理学[M].北京：人民军医出版社,2004：111.
[35]吴君.基于β3-AR探讨缩泉丸补肾缩尿的作用机制[D].广州中医药大学博十研究生论文,2010年.
[36]孙大业,郭艳林,马力更.细胞信号转导[M].第二版.北京：科学出版社,2000：55-65.
[37]Krupinski J, Coussen F, Bakalyar HA, et al.Adenylyl cyclasebamino acidsequence: possible channel-or transporter like structure.Science,1989,244:1558-1564.
[38]Wallis RM, Napier CM.Muscarinic antagonists in development for disorders of smooth muscle function.Life Sci 1999; 64(6-7):395-401.
[39]Andersson KE. Neurotransmitters and neuroreceptors in the lower urinary tract [J].Curr Opin Obstet Gynecol,1996, Oct; 8(5):361-365.
[40]Eglen RM, Reddy H, Watson N, Challiss RA.Muscarinic acetylcholine receptor subtypes in smooth muscle. Trends Pharmacol Sci 1994 Apr; 15(4):114-119.
[41]吴清和,吴君,黄萍,等.缩泉丸对自然衰老大鼠逼尿肌AC-cAMP通路的影响[J].广州中医药大学学报.2011；28(5)：512-515.
[42]龚国清.小鼠衰老模型研究[J].中国药科大报,1991；22(2)：101.
[43]樊新荣,黄贵华,朱文锋.太阴病脾虚寒湿证大鼠模型的建立与机理探讨[J].中华中医药学刊,2008；26(1)：58-60.
[44]谭莹,黄萍,操红缨.缩泉胶囊对业急性衰老大鼠逼尿肌功能的影响[J].中国中药杂志.2010；35(23)：3207-3210.
[45]姜勇,罗胜秋.细胞信号转导的分子基础和功能调控[M].北京：科学出版社,2005：10.
[46]谭莹,黄萍,石孟,等.缩泉丸对业急性哀老大鼠逼尿肌Collagen I型、Ⅲ型胶原蛋白的mRNA及蛋白表达的影响[J].中国实验方剂学杂志,2010；16(15)：126
[47]宋波.有关逼尿肌功能的理解和研究[J].临床泌尿外科杂志,2004；19(11)：641-642.
[48]宋波.逼尿肌兴奋性及逼尿肌不稳定[J].第三军医大学学报,2003；25(22)：1969-1971.
[49]何地英,瞿颂义.药物对膀胱逼尿肌的影响[J].兰州医学院学报,1996；22(1)：27-28.
[50]魏保生.医学免疫学[M].北京：科学出版社,2004：120.
[51]唐旭东,麻柔,刘锋,等.再生障碍性贫血CD4+/CD8+T亚群的CD45RA+/RO+表达与中医辨证分型的相关性研究[J].四川中医.2006；24(1)：14-16.
[52]陈治水.加味缩泉饮治疗流行性出血.热多尿期患者35例[J].上海中医.1988；(5)：28.
[53]刘和义.缩泉丸加味治疗小儿尿频症30例[J].湖北中医杂志,1995；17(1)；5.
[54]单明义.桑螵蛸散加缩泉丸治疗精神紧张性尿频6例体会[J].甘肃中医,2000；13(1)：22.
[55]陈霖.缩泉固脬汤治疗小儿神经性尿频34例[J].光明中医,2010；25(6)：970.
[56]穆莉芳,马文旭,赵红立.缩泉胶囊配合心理干预治疗小儿神经性尿频40例[J].中医儿科杂志,2007；3(5)：41-42.
[57]沈振欧,李成芳.缩泉散治疗小儿神经性32例[J].吉林中医药,1998；(2)：28.
[58]陈刚,莫松柳,张浒.缩泉饮联合硝苯吡啶治疗小儿神经性尿频疗效观察[J].中国基层医药,2006；13(7)：1199-1200.
[59]陈宝生,徐佩英,应丽君,等.缩泉丸治疗功能性尿火禁临床观察[J].上海医药,1998；19(10)：12.
[60]肖宝兰,杜彩兰,袁秀琼.缩泉丸汤加味治疗老年尿失禁11例体会[J].交通医学,2001；15(3)：339-343.
[61]钱玉琴,加味缩泉丸治疗劳淋35例[J].实用中医药杂志,2002；18(11)：12.
[62]翟栋,刘真,冉云.金樱固肾缩泉汤治疗混合性尿失禁疗效观察[J].中国社区医师,2009；25(14)：42.
[63]阮时宝,魏亚东,苑述刚,等.缩泉固尿合剂治疗老年女性尿失禁28例临床观察[J].中国中西医结合杂志,2009；29(9)：852.
[64]史雷平.益气缩泉饮加功能锻炼治疗张力性尿失禁98例[J].中国民间疗法,2006；14(6)：47-48.
[65]杨廷安.桂附地黄丸合缩泉丸加减治疗顽固性遗尿例析[J].实用中医内科杂志,2002；16(3)：158.
[66]王素平.桑螵蛸散合缩泉丸治疗小儿遗尿症[J].天津中医药,2003；20(2)：13.
[67]于冬梅,王依新.缩泉丸加味治疗小儿遗尿证[J].吉林中医药,2004；24(6)：25.
[68]刁永红.缩泉丸加味方治疗小儿遗尿58例[J].江西中医药,2004；35(2)：23.
[69]赵秀艳,孙智华.桑螵蛸散合缩泉丸加减治疗小儿遗尿28例[J].中华当代医学,2005；3(1)：72.
[70]陈丁丁,蒋伟.醒脑缩泉汤配合穴位敷贴治疗小儿遗尿86例[J].河北中医,2009；31(4)：536.
[71]周怡.健脾缩泉汤治疗小儿遗尿症52例[J].浙江中医杂志,2009,44(12)：887.
[72]乔学军王海燕.缩泉丸合香砂六君子汤加减治疗小儿原发性遗尿16例[J].山东中医杂志,2010；9(2)：100-101.
[73]陈君江.自拟益肾缩泉汤治疗小儿遗尿症30例临床观察[J].中国中医药科技,2011；18(2)：133.
[74]王文才.验方缩泉止溺汤治疗小儿遗尿症的分析[J].内蒙古中医药,2011；(23)：20-21.
[75]张红旗,李全香.缩泉丸合桑螵蛸散加减治疗肾虚型小儿遗尿尿频39例[J].中国民间疗法,2011；19(11)：34-35.
[76]朱沛冉.缩泉丸加味治疗尿崩症[J].四川中医,1987；(4)：30.
[77]叶枫.六味地黄汤合缩泉丸治疗尿崩症[J].中医研究,2002；15(4)：59.
[78]杜玉斌.叶氏固精缩泉方治愈尿崩症[J].中国中西医结合肾病杂志,2003；4(1)：58-59.
[79]叶青.补肾缩泉汤治疗老年男性夜尿增多24例[J].河北中医,2006；28(3)：184-185.
[80]张四雄.加味缩泉丸(汤)药蛋治疗小儿夜尿症41例疗效观察[.J].当代医师杂志,1997；2(5)：48.
[81]陶红.缩泉丸加减治疗尿道综合征[J].四川中医,1998；16(10)：30.
[82]李兆兵.通涩并用治疗小儿尿道综合征42例[J].新中医,2004；36(11)：63-66.
[83]杨开来.缩泉丸加味治疗尿道综合征48例[J].湖北中医杂志,2006；28(4)：39-41.
[84]刘本友.缩泉丸合六一散加减治疗慢性前列腺炎24例[J].中国民间疗法,2002；10(3)：52.
[85]刘建英.金锁固金丸与缩泉九新用[J].家庭中医药,2006；10(10)：60-61.
[86]吴国君,朱丽萍,易正辉等.缩泉丸治疗氯氮平所致流涎的疗效分析[J].四川精神卫生；2002：15(2)：82-83.
[87]汤景文.缩泉丸治疗氯氮平所致流涎42例报告[J].江西中医学院学报,2000；12(2)：56.
[88]朱继先.加味缩泉丸治疗多涕症38例[J].山西中医,2001；17(3)：37.
[89]袁怀同,赵爱霞.玉屏风散合缩泉丸治疗小儿多涕27例[J].新中医,1998；30(9)：41.
[90]李东辉,姜慧强,全德君.缩泉丸配合点穴治疗老年性迎风冷泪症[J]吉林中医药,1999；(4)：25.
[91]张海.缩泉丸加味治疗冷泪症180例[J].实用中医药杂志,2001；17(10)：10.
[92]钱聚义.缩泉丸加味妇科运用举隅[J].天津中医,1998；15(4)：186.
[93]黄腾蛟,林垣垣.缩泉丸的临床应用[J].福建中医药,1991；22(5)：41.
[94]张宏俊.缩泉丸加味治滑胎[J].江苏中医,1989；(1)：38.
[95]周广田.缩泉丸临床新用[J].内蒙古中医药,2005；(1)：27-28.
[96]张忠富.加味缩泉丸治愈双侧肾积水[J].中国社医医师,2002；4(3)：38-39.
[97]蔡震宇.缩泉丸治疗逆行射精2例[J].男科学报,1999；9(5)：183-184.
[98]刘春勇.加味缩泉丸治疗便秘1例[J].河北中医,2001；7(7)：497-500.
[99]成令忠.组织学与胚胎学[M].北京：人民卫生出版社,1978：212-213.
[100]金锡御,宋波.临床尿动力学[M].北京；人民卫生出版社,2002：6-13.
[101]何地英,瞿颂义.膀胱逼尿肌的组织及生理学概述[J].西北民族大学学报(自然科学版),2005；26(60)：82-85.
[102]Dixon JS.Ultrastructure of smooth muscle cells in the urinary system.Smooth Muscle[J]. Research,1990,1(1):160.
[103]张玉海.膀胱排尿功能障碍[M].北京：人民卫生出版社,2000：19-21.
[104]Igawa Y,Yamazaki Y,Takeda H,et al. Function and molecule or biological evidc-nce For possible β3-adrenoceptor in the human detrusor musle.Br J Pharmacol,2000,126: 819-825.
[105]HegdeSS,ChoPPinA,BonhansD,et al.eral.Functional role of M2 and M3 muscarinic receptors in the urinary bladder of rats in vitro and in vivo.Br J Phar,1997; 120:1409-1411.
[106]Rozec B.and Cauthier,C.Beta3-adrenoceptors in the cardiovascular system:putative roles in human pathologies.Pharmacol Ther,2006,111(3):652-673.
[107]刘才平,邓杰,李勤耕.选择性β3肾上腺素受体激动剂研究进展[J].中国新药杂志2007；16(8)：585-591.
[108]Fujimura T, Tamura K, Tsutsumi T,et al. Expression and possible functional role of the beta3-adrenoceptor in human and rat detrusor muscle[J]. J Urol,1999,161:680-685.
[109]李振华,马智勇,李刚,等.肾上腺素能β受体在人膀胱逼尿肌中作用的年龄相关性研究[J].中华老年多器官疾病杂志,2004；3(3)：180.
[110]Hall IP.Second messenger-ion channels pharmacology of airway smooth muscle. EurRespirJ,2000;15:1120-1127.
[111]曹文峰,宋波,陈昭颉,等.β3-AR介导逼尿肌舒张信号转导机制的实验研究[J].解放军医学杂志,2004；29(6)：516-518.
[112]Takeda H, Yamazaki Y, Akahane M, et al:Role of the β3-adrenoceptor in urine storage in the rat:comparison between the selective β3-adrenoceptor agonist, CL316,243 and various smooth muscle relaxants.[J].J Pharmacol Exp Ther.,2000,293:939-945.
[113]熊恩庆,赵新鸿,宋波.老龄大鼠膀胱逼尿肌对神经递质反应的研究[J].第三军医大学学报,2003；25(22)：1979-1981.
[114]曹文峰,宋波,金锡御,等.肾上腺素β 3-受体与梗阻后逼尿肌不稳定关系的实验研究[J].中华泌尿外科杂志,2004；25(5)：325-327.
[115]Nishimoto T,Latifpour J, Wheeler MA,et al.Age-dependent alterations in beta-adrenergic responsiveness of rat detrusor smooth muscle[J].J Urol,1995,153:1701-1705.
[116]赵新鸿,熊恩庆,宋波,等.老年大鼠膀胱逼尿肌舒张功能改变的神经机制研究[J].华北国防医药,2004；16(1)：36-38.
[117]曹文峰,宋波,金锡御,等.逼尿肌不稳定大鼠肾上腺素β及β3受体密度和功能变化的研究[J].解放军医学杂志,2003；28(2)：134-136.
[118]李振华,马智勇,李刚,等.肾上腺素能β受体在人体膀胱逼尿肌中作用的年龄相关性研究[J].中华老年多器官疾病杂志,2004；3(3)：180.
[119]Hulme EC,Lu ZL,Saldanha JW,et al.Structure and activation of muscarinic acetylcholine rcceptors[J].Biochem Soc Trans.2003;31 (Pt 1):29-34.
[120]Wess J,Blin M,Mutsehler E,et al.Muscarinic acetylcholine receptors:structural basis of ligand b ingding and G protein coupling[J].Life Sci,1995;56(11/12):915-922.
[121]Sellers DJ. M3 muscarinic receptors but not M2 mediate contraction of the porcine detrusor muscle in vitro.J Auton Pharmmol,2000,20(3):171-176.
[122]左伋,医学分子细胞生物学[M].复旦大学出版社,2005：97.
[123]腾若冰.M2、M3受体与膀胱逼尿肌关系的研究进展[J].国际泌尿系统杂志,2006；26(5)：676-680.
[124]Felder CC. Muscarinie acetylcholine receptors:signal transduction through multiple effectors[J]. FASEB J,1995,9(8):619-625.
[125]孙科,郭正东.M受体-G蛋白信号转导途径研究进展[J].国际药学研究杂志,2008；35(2)：87-91.
[126]Racke K,Juergens UR,Matthiesen S.Control by cholinergic mechanisms[J]. Eur jpharm-acol,2006,533(1-3):57-68.
[127]Fleichman M,Schneider T,Fetscher C,eral.Signal Transduction Underlying Carbacholind-uced Contraction of Rat Urinary Bladder.Ⅱ.Protein Kinase.J Pharmacol Expther,2004:(308):54-58.
[128]李新,宋波,李益凡,等.M受体及其业型与逼尿肌功能的关系研究[J].第三军医大学学报.2002；24(2)：164-166.
[129]李新,宋波,金锡御,等.M受体及其业型与脊髓横断后逼尿肌反射亢进关系的研究[J].中华泌尿外科杂志,2002；23(8)：501-503.
[130]Ikeda K,Kobayashi s,Suzuki M et al. M3 receptor antagonism by the novel Antimuscarinic agent solifenacin in the urinary bladder and salivary gland.Naunyn Schmiedebergs Arch.Pham.acol 2002;366:97-103.
[131]高宏飞,王东文,双卫兵.2型糖尿病大鼠膀胱M3受体改变的实验研究[J].山西医药杂志,2006；35(6)：475-477.
[132]王新民,薛萌.胰岛素治疗对糖尿病大鼠膀胱M2、M3乙酰胆碱受体nRNA表达的影响[J].中国糖尿病杂志,2009；17(9)：705-706.
[133]卢根生,宋波,熊恩庆,等.大鼠不稳定膀胱与MR业型密度改变关系的实验研究[J].第三军医大学学报,2002；24(12)：1437-1438.
[134]Tammela TL.Temporal changes in micturition and bladder contractility after sucrose diuresis and streptozotocin-induced diabetes mellilus in rats[J].Urol,1995; 153(6):2014-2120.
[135]Lluel PM, Barras,Palea S. Cholinergic and purinergic contribution to the micturition reflex in conscious rats with long-term bladder outlet obstruction. [J].Neurourol Urodyn, 2002;21(2):142-53.
[136]Zinner N,Susset J,Gittelmanm M,Arguinzoniz M,Rekeda L,Haab F. Effieaey, tolerability and safety of darifenacin,an M3 seleetive receptor antagonist:an investigation of warning time in Patients with OAB.Int.J.Clin.Pract.2006:60:119-126.
[137]姚泰.生理学[M].北京：人民卫生出版社,2001：275-312.
[138]周逢海,王养民,宋波,等.膀胱逼尿肌细胞的培养及鉴定[J].西北国防医学杂志2006,17(1)：5-7.
[139]朱详,王平,刘屹立,等.逼尿肌细胞的原代培养及鉴定[J].临床军医杂志,2010；10(5)：699-701.
[140]宋今丹.组织和细胞培养技术[M].北京：人民卫生出版社,2004：5.
[141]章静波,徐存拴译.动物细胞培养[M].北京：科学出版社,2002：16.
[142]Wang P,Luthin GR,Ruggieri MR.Muscarinic acetylcholine receptor subtype mediation urinary bladder contractility and coupling TO GTP binding proteins.J Pharmacol Exper Ther,1995,273:959-966.
[143]Somogyi GT,De Groat WC.Function signal transduction mechanisms and plasticity of prisynaptic muscarinic receptor in the urinary bladde.Life Sci,1999,64:411-418.
[144]KriefS,Lonnqvist F,Raimbault,S,et al.Tissue distribution of beta 3-adrenergicreceptor mRNA in man.J-Clin-Invest.1993 Jan;91(1):344-9.
[145]Penelope A.Longhurst M.Pharmacological characterization of β-adrenoceptors mediating relaxation of the rat urinary bladder in vitro.Br.J Pharmacol,1999,127:1744-1750.
[146]Kolta MG,Wallace LJ,Gerald MC.Age-related changes in sensitivity of rat urinary bladder to autonomic agents.Mech Ageing Dev,1984,27(2):183-188.
[147]Wide E,Lehto M,Kanninen T,et al: Association of a polymorphism in the P3-adrenergic-receptor gene with features of the insulin resistance syndrome in Finns.N Engl J Med,1995;333:348-51.
[148]Konkar AA, Zhu Z, and Granneman JG:Aryloxypropanolamine and catccholamine ligand interactions with the (β1-adrenergic receptor:evidence for interaction with distinct conformations of β2-adrenergic receptors. J Pharmacol Exp Ther,2000; 294:923-932.
[149]卢根生,宋波,金锡御,等.M3R介导逼尿肌细胞收缩与信使分子IP3关系的实验研究[J].中国现代医学杂志,2002；12(24)：33-34.
[150]卢根生,宋波,金锡御,等.毒蕈碱受体亚型介导逼尿肌细胞收缩与IP3关系的实验研究[J].2003；24(10)：678-680.
[151]王军,梁颖,庄仕华,等.膀胱醒阻大鼠逼尿肌不稳定与三磷酸肌醇关系的实验研究[J].新疆医科大学学报,2007；30(8)：803-806.
[152]林明群,张宗梁.蛋白激酶A(PKA)介导的信号通路：正调节还是负调节[J].科学通报,1999；44(17)：1793-1803.
[153]张向阳.激光共聚焦扫描显微镜与流式细胞仪在医学研究中的比较[J].邯郸医学高等专科学校学报,2002；15(6)：668-670.
[154]黄娅林,赵鹏,程介士.共聚焦激光扫描显微镜技术在研究大鼠海马神经元胞内钙超载中的应用[J].复旦学报(医学版),2005,32(1)：98-99.
[155]曹文峰.钙离子在逼尿肌收缩机制中的作用[J],西南国防医药,2005；15(1)：103-105.
[156]卢根生,宋波,金锡御,等.M3R介导逼尿肌细胞收缩与信使分子IP3关系的实验研究[J].中国现代医学杂志.2002；12(24)：33-34.
[157]王军,梁颖,庄仕华,等.膀胱梗阻大鼠逼尿肌不稳定与三磷酸肌醇关系的实验研究[J].新疆医科大学学报.2007；30(8)：803-806.
[158]卢根生,任静,宋波,等Ca2+-CaM在M3R介导逼尿肌细胞收缩中作用的实验研究[J].第三军医大学学报,2003；25(23)：2123.
[159]李家旭,孙大业.生物细胞中钙调素分布研究及其意义[J].细胞生物学杂志,1991；1：1-3.
[160]王军,梁颖,庄仕华,等.三磷酸肌醇对大鼠逼尿肌细胞内钙影响的激光共聚焦显微镜观察[J].新疆医科大学学报.2007；30(8)：809-812.
[161]陆金宝,周如倩.老年人肾虚及其证型的调查研究[J].上海中医药大学学报,2002；16(1)：22-23.
[162]冯淑香,刘耀明,董俊兴.中药益智仁化学成分与药理研究进展[J].现代中药研究与实践,2003；17(5)：58-62
[163]王军伟,阮冰.乌药的植化及药理研究概况[J].浙江中医杂志,2006；41(11)：675-678
[164]何海玲,单承莺,张卫明,等.山药研究进展[J].中国野生植物资源,2006；25(6)：1-7
[2]谭莹.基于逼尿肌胶原蛋白及神经受体探讨缩泉丸作用机制[D].广州中医药大学博士研究生论文,2009年.
[3]操红缨.基于AQP_2研究缩泉丸补肾缩尿的机理[D].广州中医药大学博士研究生论文,2008年
[4]王米渠,胡聪.试论肾主水司二阴并及肾体[J].片肃中医,1998,11(5)：1-4.
[5]李奕祺.“精水合一”是肾藏象的认识基础浅识[J].中医药学刊,2004,22(2)：302
[6]陈明,龙子江,卞海.补肾缩泉胶囊对老年性夜尿增多症模型及老年大鼠尿量的影响[J].中国中医药科技,2007；14(5)：337-339
[7]吕圭源.药理学(第2版)[M].北京：中国中医药出版社,2007：149.
[8]朱大年.生理学(第7版)[M].北京：人民卫生出版社,2008：212-230.
[9]操红缨,吴清和,黄萍,等.缩泉丸对肾阳虚多尿大鼠尿BUN, Cr、Na+、K+、Cl离子浓度的影响[J].中医药临床杂志,2009；21(2)：117.
[10]操红缨,吴清和,黄萍,等.缩泉丸对肾阳虚多尿大鼠肾脏病理的影响[J].中国现代药物应用.2009；3(14)：59-60.
[11]刘春蓓,胡伟新.肾素-血管紧张素系统：新成员,新效应[J].肾脏病与透析肾移植杂志,2005；14(4)：349-355.
[12]吴玉付,李醒三.肾素-血管紧张素-醛固酮系统在心血管病中的作用[J].医学文选,2007；24(6)：1023-1027.
[13]刘建萍,刘精东.肾素血管紧张素系统与糖尿病肾病相关性研究进展[J].江西医药,2008；43(8)：851-854.
[14][19] Taufiq-Ur-Rahman M,Shilpi JA,Ahmed M,et al.Preliminary Pharmacological studies on Piper chaba stem bark.J Ethnopharmacol,2005;99(2):203-209.
[15]刘进平,韩涛,浦晓东.血.管紧张素Ⅱ受体研究进展[J].福建医药杂志,2006；28(4)：97-100.
[16]叶绪英,宋卉,卢成志.葛根素注射液对自发性高血压大鼠AT1和ACE2mRNA表达的影响[J].中国中西医结合杂志,2008；28(9)：824-828.
[17]李淑雯,吴清和,黄萍,等.缩泉丸对肾虚多尿大鼠肾脏MRmRNA及蛋白表达的影响[J].中药药理与临床,2010；26(6)：8-10.
[18]SoPhie Combet,Nancy Geffroy,Veronique Berthonaud.Correction of Age-related Polyuria by dDAVP:molecular analysis of aquaporins and urea transporters.Am J Physiol Renal Physiol,2003;284:F199-F208.
[19]Katsuya Suzuki,Ryo Hatano,Mari Michimata,et al.Residual Urinary Concentrat-ing Ability and AQP2 Expression in a Rat Model for Chronie Renal Failure. Neph-ron Physiol 2005;(99):P16-P22.
[20]操红缨,吴清和,黄萍,等.缩泉丸对肾阳虚多尿大鼠肾脏AQP-2与AVPR-V2mRNA表达的影响[J].中药材,2009；2(6)：926-928.
[21]连晓媛,陈奇.应激与中枢神经递质及HPA轴功能调节[J].国外医学·生理、病理科学与临床分册,1998；18(4)：371-374.
[22]李淑雯,吴清和,黄萍,等.腺嘌呤对大鼠下丘脑-垂体-肾上腺皮质(HPA)轴的影响[J].时珍国医国药,2010；21(9)：2358.
[23]操红缨,吴清和,黄萍,等.缩泉丸对肾阳虚多尿大鼠内分泌及免疫功能的影响[J].中药新药与临床药理.2009；20(4)：323-326.
[24]Jun-Ho La,Tae-Sik Sung,Hyun-Ju Kim.Peripheral corticotropin releasing Hormone mediates Post-inflammatory visceral hypersensitivity in rats. (?)世界胃肠病学杂志《英文版》,2008；14(5)：731-736.
[25]郑小伟,包素珍,李荣群,等.二仙汤对肾阳虚大鼠垂体ACTH基因表达的影响[J].中国医药学报,2003；18(12)：716-719.
[26]张秀珍.内分泌代谢疾病与肾脏[M].上海：复旦大学出版社,2004：50.
[27]赖凌云,顾勇.醛固酮的新认识[J].国外医学·生理、病理科学与临床分册.2002；22(4)：347-349.
[28]李晓东,唐德集,刘华锋,等.醛固酮与肾脏疾病[J].国外医学·生理、病理科学与临床分册,2005；25(3)：232-236.
[29]King LS,Kozono D,Agre P:From structure to disease.The evolving tale of Aquaporin biology.Nat Rev Mol Cell Biol 2004:5:687-698.
[30]Klussmann E,Maric K,Wiesner B,et al.Protein kinase A anchoring Proteins areRequired for vasoPressin-mediated translocation of aquaporin-2 into cell membranes of renal Principal cells.J BiolCheml999:274:4934-4938.
[31]S.Rttig,J.Frokier.Basis and therapeutical rationale of the urinary Concentrating mechanism.Int J Clin Pract,2007,61(Supp1·155),2-7.
[32]操红缨,吴清和,黄萍,等.缩泉九对肾阳虚多尿模型大鼠血cAMP和ALD水平的影响[J].新中医,2009；41(10)：101.
[33]于赫,郭丽新.肾阳虚证的生物化学与分子生物学研究进展[J].中医药信息,2005；22(6)：41-44.
[34]林茂樟,临床肾脏生理学[M].北京：人民军医出版社,2004：111.
[35]吴君.基于β3-AR探讨缩泉丸补肾缩尿的作用机制[D].广州中医药大学博十研究生论文,2010年.
[36]孙大业,郭艳林,马力更.细胞信号转导[M].第二版.北京：科学出版社,2000：55-65.
[37]Krupinski J, Coussen F, Bakalyar HA, et al.Adenylyl cyclasebamino acidsequence: possible channel-or transporter like structure.Science,1989,244:1558-1564.
[38]Wallis RM, Napier CM.Muscarinic antagonists in development for disorders of smooth muscle function.Life Sci 1999; 64(6-7):395-401.
[39]Andersson KE. Neurotransmitters and neuroreceptors in the lower urinary tract [J].Curr Opin Obstet Gynecol,1996, Oct; 8(5):361-365.
[40]Eglen RM, Reddy H, Watson N, Challiss RA.Muscarinic acetylcholine receptor subtypes in smooth muscle. Trends Pharmacol Sci 1994 Apr; 15(4):114-119.
[41]吴清和,吴君,黄萍,等.缩泉丸对自然衰老大鼠逼尿肌AC-cAMP通路的影响[J].广州中医药大学学报.2011；28(5)：512-515.
[42]龚国清.小鼠衰老模型研究[J].中国药科大报,1991；22(2)：101.
[43]樊新荣,黄贵华,朱文锋.太阴病脾虚寒湿证大鼠模型的建立与机理探讨[J].中华中医药学刊,2008；26(1)：58-60.
[44]谭莹,黄萍,操红缨.缩泉胶囊对业急性衰老大鼠逼尿肌功能的影响[J].中国中药杂志.2010；35(23)：3207-3210.
[45]姜勇,罗胜秋.细胞信号转导的分子基础和功能调控[M].北京：科学出版社,2005：10.
[46]谭莹,黄萍,石孟,等.缩泉丸对业急性哀老大鼠逼尿肌Collagen I型、Ⅲ型胶原蛋白的mRNA及蛋白表达的影响[J].中国实验方剂学杂志,2010；16(15)：126
[47]宋波.有关逼尿肌功能的理解和研究[J].临床泌尿外科杂志,2004；19(11)：641-642.
[48]宋波.逼尿肌兴奋性及逼尿肌不稳定[J].第三军医大学学报,2003；25(22)：1969-1971.
[49]何地英,瞿颂义.药物对膀胱逼尿肌的影响[J].兰州医学院学报,1996；22(1)：27-28.
[50]魏保生.医学免疫学[M].北京：科学出版社,2004：120.
[51]唐旭东,麻柔,刘锋,等.再生障碍性贫血CD4+/CD8+T亚群的CD45RA+/RO+表达与中医辨证分型的相关性研究[J].四川中医.2006；24(1)：14-16.
[52]陈治水.加味缩泉饮治疗流行性出血.热多尿期患者35例[J].上海中医.1988；(5)：28.
[53]刘和义.缩泉丸加味治疗小儿尿频症30例[J].湖北中医杂志,1995；17(1)；5.
[54]单明义.桑螵蛸散加缩泉丸治疗精神紧张性尿频6例体会[J].甘肃中医,2000；13(1)：22.
[55]陈霖.缩泉固脬汤治疗小儿神经性尿频34例[J].光明中医,2010；25(6)：970.
[56]穆莉芳,马文旭,赵红立.缩泉胶囊配合心理干预治疗小儿神经性尿频40例[J].中医儿科杂志,2007；3(5)：41-42.
[57]沈振欧,李成芳.缩泉散治疗小儿神经性32例[J].吉林中医药,1998；(2)：28.
[58]陈刚,莫松柳,张浒.缩泉饮联合硝苯吡啶治疗小儿神经性尿频疗效观察[J].中国基层医药,2006；13(7)：1199-1200.
[59]陈宝生,徐佩英,应丽君,等.缩泉丸治疗功能性尿火禁临床观察[J].上海医药,1998；19(10)：12.
[60]肖宝兰,杜彩兰,袁秀琼.缩泉丸汤加味治疗老年尿失禁11例体会[J].交通医学,2001；15(3)：339-343.
[61]钱玉琴,加味缩泉丸治疗劳淋35例[J].实用中医药杂志,2002；18(11)：12.
[62]翟栋,刘真,冉云.金樱固肾缩泉汤治疗混合性尿失禁疗效观察[J].中国社区医师,2009；25(14)：42.
[63]阮时宝,魏亚东,苑述刚,等.缩泉固尿合剂治疗老年女性尿失禁28例临床观察[J].中国中西医结合杂志,2009；29(9)：852.
[64]史雷平.益气缩泉饮加功能锻炼治疗张力性尿失禁98例[J].中国民间疗法,2006；14(6)：47-48.
[65]杨廷安.桂附地黄丸合缩泉丸加减治疗顽固性遗尿例析[J].实用中医内科杂志,2002；16(3)：158.
[66]王素平.桑螵蛸散合缩泉丸治疗小儿遗尿症[J].天津中医药,2003；20(2)：13.
[67]于冬梅,王依新.缩泉丸加味治疗小儿遗尿证[J].吉林中医药,2004；24(6)：25.
[68]刁永红.缩泉丸加味方治疗小儿遗尿58例[J].江西中医药,2004；35(2)：23.
[69]赵秀艳,孙智华.桑螵蛸散合缩泉丸加减治疗小儿遗尿28例[J].中华当代医学,2005；3(1)：72.
[70]陈丁丁,蒋伟.醒脑缩泉汤配合穴位敷贴治疗小儿遗尿86例[J].河北中医,2009；31(4)：536.
[71]周怡.健脾缩泉汤治疗小儿遗尿症52例[J].浙江中医杂志,2009,44(12)：887.
[72]乔学军王海燕.缩泉丸合香砂六君子汤加减治疗小儿原发性遗尿16例[J].山东中医杂志,2010；9(2)：100-101.
[73]陈君江.自拟益肾缩泉汤治疗小儿遗尿症30例临床观察[J].中国中医药科技,2011；18(2)：133.
[74]王文才.验方缩泉止溺汤治疗小儿遗尿症的分析[J].内蒙古中医药,2011；(23)：20-21.
[75]张红旗,李全香.缩泉丸合桑螵蛸散加减治疗肾虚型小儿遗尿尿频39例[J].中国民间疗法,2011；19(11)：34-35.
[76]朱沛冉.缩泉丸加味治疗尿崩症[J].四川中医,1987；(4)：30.
[77]叶枫.六味地黄汤合缩泉丸治疗尿崩症[J].中医研究,2002；15(4)：59.
[78]杜玉斌.叶氏固精缩泉方治愈尿崩症[J].中国中西医结合肾病杂志,2003；4(1)：58-59.
[79]叶青.补肾缩泉汤治疗老年男性夜尿增多24例[J].河北中医,2006；28(3)：184-185.
[80]张四雄.加味缩泉丸(汤)药蛋治疗小儿夜尿症41例疗效观察[.J].当代医师杂志,1997；2(5)：48.
[81]陶红.缩泉丸加减治疗尿道综合征[J].四川中医,1998；16(10)：30.
[82]李兆兵.通涩并用治疗小儿尿道综合征42例[J].新中医,2004；36(11)：63-66.
[83]杨开来.缩泉丸加味治疗尿道综合征48例[J].湖北中医杂志,2006；28(4)：39-41.
[84]刘本友.缩泉丸合六一散加减治疗慢性前列腺炎24例[J].中国民间疗法,2002；10(3)：52.
[85]刘建英.金锁固金丸与缩泉九新用[J].家庭中医药,2006；10(10)：60-61.
[86]吴国君,朱丽萍,易正辉等.缩泉丸治疗氯氮平所致流涎的疗效分析[J].四川精神卫生；2002：15(2)：82-83.
[87]汤景文.缩泉丸治疗氯氮平所致流涎42例报告[J].江西中医学院学报,2000；12(2)：56.
[88]朱继先.加味缩泉丸治疗多涕症38例[J].山西中医,2001；17(3)：37.
[89]袁怀同,赵爱霞.玉屏风散合缩泉丸治疗小儿多涕27例[J].新中医,1998；30(9)：41.
[90]李东辉,姜慧强,全德君.缩泉丸配合点穴治疗老年性迎风冷泪症[J]吉林中医药,1999；(4)：25.
[91]张海.缩泉丸加味治疗冷泪症180例[J].实用中医药杂志,2001；17(10)：10.
[92]钱聚义.缩泉丸加味妇科运用举隅[J].天津中医,1998；15(4)：186.
[93]黄腾蛟,林垣垣.缩泉丸的临床应用[J].福建中医药,1991；22(5)：41.
[94]张宏俊.缩泉丸加味治滑胎[J].江苏中医,1989；(1)：38.
[95]周广田.缩泉丸临床新用[J].内蒙古中医药,2005；(1)：27-28.
[96]张忠富.加味缩泉丸治愈双侧肾积水[J].中国社医医师,2002；4(3)：38-39.
[97]蔡震宇.缩泉丸治疗逆行射精2例[J].男科学报,1999；9(5)：183-184.
[98]刘春勇.加味缩泉丸治疗便秘1例[J].河北中医,2001；7(7)：497-500.
[99]成令忠.组织学与胚胎学[M].北京：人民卫生出版社,1978：212-213.
[100]金锡御,宋波.临床尿动力学[M].北京；人民卫生出版社,2002：6-13.
[101]何地英,瞿颂义.膀胱逼尿肌的组织及生理学概述[J].西北民族大学学报(自然科学版),2005；26(60)：82-85.
[102]Dixon JS.Ultrastructure of smooth muscle cells in the urinary system.Smooth Muscle[J]. Research,1990,1(1):160.
[103]张玉海.膀胱排尿功能障碍[M].北京：人民卫生出版社,2000：19-21.
[104]Igawa Y,Yamazaki Y,Takeda H,et al. Function and molecule or biological evidc-nce For possible β3-adrenoceptor in the human detrusor musle.Br J Pharmacol,2000,126: 819-825.
[105]HegdeSS,ChoPPinA,BonhansD,et al.eral.Functional role of M2 and M3 muscarinic receptors in the urinary bladder of rats in vitro and in vivo.Br J Phar,1997; 120:1409-1411.
[106]Rozec B.and Cauthier,C.Beta3-adrenoceptors in the cardiovascular system:putative roles in human pathologies.Pharmacol Ther,2006,111(3):652-673.
[107]刘才平,邓杰,李勤耕.选择性β3肾上腺素受体激动剂研究进展[J].中国新药杂志2007；16(8)：585-591.
[108]Fujimura T, Tamura K, Tsutsumi T,et al. Expression and possible functional role of the beta3-adrenoceptor in human and rat detrusor muscle[J]. J Urol,1999,161:680-685.
[109]李振华,马智勇,李刚,等.肾上腺素能β受体在人膀胱逼尿肌中作用的年龄相关性研究[J].中华老年多器官疾病杂志,2004；3(3)：180.
[110]Hall IP.Second messenger-ion channels pharmacology of airway smooth muscle. EurRespirJ,2000;15:1120-1127.
[111]曹文峰,宋波,陈昭颉,等.β3-AR介导逼尿肌舒张信号转导机制的实验研究[J].解放军医学杂志,2004；29(6)：516-518.
[112]Takeda H, Yamazaki Y, Akahane M, et al:Role of the β3-adrenoceptor in urine storage in the rat:comparison between the selective β3-adrenoceptor agonist, CL316,243 and various smooth muscle relaxants.[J].J Pharmacol Exp Ther.,2000,293:939-945.
[113]熊恩庆,赵新鸿,宋波.老龄大鼠膀胱逼尿肌对神经递质反应的研究[J].第三军医大学学报,2003；25(22)：1979-1981.
[114]曹文峰,宋波,金锡御,等.肾上腺素β 3-受体与梗阻后逼尿肌不稳定关系的实验研究[J].中华泌尿外科杂志,2004；25(5)：325-327.
[115]Nishimoto T,Latifpour J, Wheeler MA,et al.Age-dependent alterations in beta-adrenergic responsiveness of rat detrusor smooth muscle[J].J Urol,1995,153:1701-1705.
[116]赵新鸿,熊恩庆,宋波,等.老年大鼠膀胱逼尿肌舒张功能改变的神经机制研究[J].华北国防医药,2004；16(1)：36-38.
[117]曹文峰,宋波,金锡御,等.逼尿肌不稳定大鼠肾上腺素β及β3受体密度和功能变化的研究[J].解放军医学杂志,2003；28(2)：134-136.
[118]李振华,马智勇,李刚,等.肾上腺素能β受体在人体膀胱逼尿肌中作用的年龄相关性研究[J].中华老年多器官疾病杂志,2004；3(3)：180.
[119]Hulme EC,Lu ZL,Saldanha JW,et al.Structure and activation of muscarinic acetylcholine rcceptors[J].Biochem Soc Trans.2003;31 (Pt 1):29-34.
[120]Wess J,Blin M,Mutsehler E,et al.Muscarinic acetylcholine receptors:structural basis of ligand b ingding and G protein coupling[J].Life Sci,1995;56(11/12):915-922.
[121]Sellers DJ. M3 muscarinic receptors but not M2 mediate contraction of the porcine detrusor muscle in vitro.J Auton Pharmmol,2000,20(3):171-176.
[122]左伋,医学分子细胞生物学[M].复旦大学出版社,2005：97.
[123]腾若冰.M2、M3受体与膀胱逼尿肌关系的研究进展[J].国际泌尿系统杂志,2006；26(5)：676-680.
[124]Felder CC. Muscarinie acetylcholine receptors:signal transduction through multiple effectors[J]. FASEB J,1995,9(8):619-625.
[125]孙科,郭正东.M受体-G蛋白信号转导途径研究进展[J].国际药学研究杂志,2008；35(2)：87-91.
[126]Racke K,Juergens UR,Matthiesen S.Control by cholinergic mechanisms[J]. Eur jpharm-acol,2006,533(1-3):57-68.
[127]Fleichman M,Schneider T,Fetscher C,eral.Signal Transduction Underlying Carbacholind-uced Contraction of Rat Urinary Bladder.Ⅱ.Protein Kinase.J Pharmacol Expther,2004:(308):54-58.
[128]李新,宋波,李益凡,等.M受体及其业型与逼尿肌功能的关系研究[J].第三军医大学学报.2002；24(2)：164-166.
[129]李新,宋波,金锡御,等.M受体及其业型与脊髓横断后逼尿肌反射亢进关系的研究[J].中华泌尿外科杂志,2002；23(8)：501-503.
[130]Ikeda K,Kobayashi s,Suzuki M et al. M3 receptor antagonism by the novel Antimuscarinic agent solifenacin in the urinary bladder and salivary gland.Naunyn Schmiedebergs Arch.Pham.acol 2002;366:97-103.
[131]高宏飞,王东文,双卫兵.2型糖尿病大鼠膀胱M3受体改变的实验研究[J].山西医药杂志,2006；35(6)：475-477.
[132]王新民,薛萌.胰岛素治疗对糖尿病大鼠膀胱M2、M3乙酰胆碱受体nRNA表达的影响[J].中国糖尿病杂志,2009；17(9)：705-706.
[133]卢根生,宋波,熊恩庆,等.大鼠不稳定膀胱与MR业型密度改变关系的实验研究[J].第三军医大学学报,2002；24(12)：1437-1438.
[134]Tammela TL.Temporal changes in micturition and bladder contractility after sucrose diuresis and streptozotocin-induced diabetes mellilus in rats[J].Urol,1995; 153(6):2014-2120.
[135]Lluel PM, Barras,Palea S. Cholinergic and purinergic contribution to the micturition reflex in conscious rats with long-term bladder outlet obstruction. [J].Neurourol Urodyn, 2002;21(2):142-53.
[136]Zinner N,Susset J,Gittelmanm M,Arguinzoniz M,Rekeda L,Haab F. Effieaey, tolerability and safety of darifenacin,an M3 seleetive receptor antagonist:an investigation of warning time in Patients with OAB.Int.J.Clin.Pract.2006:60:119-126.
[137]姚泰.生理学[M].北京：人民卫生出版社,2001：275-312.
[138]周逢海,王养民,宋波,等.膀胱逼尿肌细胞的培养及鉴定[J].西北国防医学杂志2006,17(1)：5-7.
[139]朱详,王平,刘屹立,等.逼尿肌细胞的原代培养及鉴定[J].临床军医杂志,2010；10(5)：699-701.
[140]宋今丹.组织和细胞培养技术[M].北京：人民卫生出版社,2004：5.
[141]章静波,徐存拴译.动物细胞培养[M].北京：科学出版社,2002：16.
[142]Wang P,Luthin GR,Ruggieri MR.Muscarinic acetylcholine receptor subtype mediation urinary bladder contractility and coupling TO GTP binding proteins.J Pharmacol Exper Ther,1995,273:959-966.
[143]Somogyi GT,De Groat WC.Function signal transduction mechanisms and plasticity of prisynaptic muscarinic receptor in the urinary bladde.Life Sci,1999,64:411-418.
[144]KriefS,Lonnqvist F,Raimbault,S,et al.Tissue distribution of beta 3-adrenergicreceptor mRNA in man.J-Clin-Invest.1993 Jan;91(1):344-9.
[145]Penelope A.Longhurst M.Pharmacological characterization of β-adrenoceptors mediating relaxation of the rat urinary bladder in vitro.Br.J Pharmacol,1999,127:1744-1750.
[146]Kolta MG,Wallace LJ,Gerald MC.Age-related changes in sensitivity of rat urinary bladder to autonomic agents.Mech Ageing Dev,1984,27(2):183-188.
[147]Wide E,Lehto M,Kanninen T,et al: Association of a polymorphism in the P3-adrenergic-receptor gene with features of the insulin resistance syndrome in Finns.N Engl J Med,1995;333:348-51.
[148]Konkar AA, Zhu Z, and Granneman JG:Aryloxypropanolamine and catccholamine ligand interactions with the (β1-adrenergic receptor:evidence for interaction with distinct conformations of β2-adrenergic receptors. J Pharmacol Exp Ther,2000; 294:923-932.
[149]卢根生,宋波,金锡御,等.M3R介导逼尿肌细胞收缩与信使分子IP3关系的实验研究[J].中国现代医学杂志,2002；12(24)：33-34.
[150]卢根生,宋波,金锡御,等.毒蕈碱受体亚型介导逼尿肌细胞收缩与IP3关系的实验研究[J].2003；24(10)：678-680.
[151]王军,梁颖,庄仕华,等.膀胱醒阻大鼠逼尿肌不稳定与三磷酸肌醇关系的实验研究[J].新疆医科大学学报,2007；30(8)：803-806.
[152]林明群,张宗梁.蛋白激酶A(PKA)介导的信号通路：正调节还是负调节[J].科学通报,1999；44(17)：1793-1803.
[153]张向阳.激光共聚焦扫描显微镜与流式细胞仪在医学研究中的比较[J].邯郸医学高等专科学校学报,2002；15(6)：668-670.
[154]黄娅林,赵鹏,程介士.共聚焦激光扫描显微镜技术在研究大鼠海马神经元胞内钙超载中的应用[J].复旦学报(医学版),2005,32(1)：98-99.
[155]曹文峰.钙离子在逼尿肌收缩机制中的作用[J],西南国防医药,2005；15(1)：103-105.
[156]卢根生,宋波,金锡御,等.M3R介导逼尿肌细胞收缩与信使分子IP3关系的实验研究[J].中国现代医学杂志.2002；12(24)：33-34.
[157]王军,梁颖,庄仕华,等.膀胱梗阻大鼠逼尿肌不稳定与三磷酸肌醇关系的实验研究[J].新疆医科大学学报.2007；30(8)：803-806.
[158]卢根生,任静,宋波,等Ca2+-CaM在M3R介导逼尿肌细胞收缩中作用的实验研究[J].第三军医大学学报,2003；25(23)：2123.
[159]李家旭,孙大业.生物细胞中钙调素分布研究及其意义[J].细胞生物学杂志,1991；1：1-3.
[160]王军,梁颖,庄仕华,等.三磷酸肌醇对大鼠逼尿肌细胞内钙影响的激光共聚焦显微镜观察[J].新疆医科大学学报.2007；30(8)：809-812.
[161]陆金宝,周如倩.老年人肾虚及其证型的调查研究[J].上海中医药大学学报,2002；16(1)：22-23.
[162]冯淑香,刘耀明,董俊兴.中药益智仁化学成分与药理研究进展[J].现代中药研究与实践,2003；17(5)：58-62
[163]王军伟,阮冰.乌药的植化及药理研究概况[J].浙江中医杂志,2006；41(11)：675-678
[164]何海玲,单承莺,张卫明,等.山药研究进展[J].中国野生植物资源,2006；25(6)：1-7