Based On Detrusor Receptor To Research Regulation And Mechanism Of Suoquan Wan (SQW)

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 microscopeResults: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 rats5. 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...