Based On The Collagen Protein And Neuroreceptor Of Detrusor To Investigate Mechanisms Of SQW

The low-tract urinary symptoms(LTUS) like nocturia,urinary frequency,enuresis,urinary incontinence,etc have a highly incidence rate in the elder people.Substituted heart and cerebrovascular system,it become the critical reason to effect physical and mental health and life quality of the aged people. According study,we find out the cause of the LTUS in the aged people not only the reason of urethra,like nerve system dysfunction,blood-vascular system affection and bladder outflow obstruction,but also the age itself.The function of capsular bladder is to urinate and store.This physiological function depend detrusor of bladder,a kind of main smooth muscle of bladder, to contract and relax.Because of physical and neurogenic changes,the senescent detrusor muscle occur functional impairment of contraction and relaxation.This is the main cause of elder LTUS.The formula of Suoquanwan(SQW) is recorded in a Chinese medicine book named Furenliangfang,which collected hundreds of formulas of Chinese medicine combinations.SQW consists of three herb medicines,Radix Linderae,Radix dioscoreae,and alpinia oxyphylla Miq. The function of SQW is to warm kidney,stop polyuria and hold excessive urination.This research duplicate the animal model of senile hyperdiuresis and observes the effect of arresting polyuria,collagen protein of musculus detrusor vesicae,neuroceptor related gene and regulatory protein related expression.This study from different Angles,like the molecular mechanism and the occurrence and development of polyuria,search for the mechanism of the formula SQW.Objective:On the basis of duplicating D-galactose mimetic aging animal model and exploring its mechanism of hyperdiuresis,observed the pharmacological action and molecular mechanism of SQW.Methods:Investigated the urination of D-galactose mimetic aging animal model, including total volume and ionic concentration of urine during 24h;We also observed the incubation period and frequency of urine within 6h after water load.The pharmacological mechanisms of SQW were investigated in following aspects:Investigated the hormones(including:ADH,ALD,ANP) and enzymes (including:ATPase,SDH) which influence the production and excretion of urine;the detrusor strips of each rat was suspended in a thermostatically controlled organ bath.And their spontaneous contraction frequency,reaction tension to stress and response intensity to electric stimuli were determined and statistically analyzed;The reactivity of urinary detrusor strips of each rat to different concentrations of ISO and ATP was measured respectively using an in vitro tissue bath technique;The pathological changes of kidney and bladder in rats were observed with HE,Masson,PAS dying method before and after SQW administration.The influence of SQW on expression of collagenâ… mRNA,collagenâ…¢mRNA,P2X₂ mRNA,β₃-AR mRNA in urinary detrusor strips of O-galactose mimetic aging animal model by RT-PCR;The influence of SQW on the in situ and protein expressions of collagenâ… ,collagenâ…¢,P2X₂,β₃-AR in urinary detrusor strips of polyuria model by immunohistochemistry method.Results:1,The mechanism of polyuria for D-galactose mimetic aging modelThe animal models have characteristics of polyuria.Total volume of urine during 24h increased.The incubation periods shorten and the frequency of urine increased within 6h after water load.There are about some reasons that caused these symptoms.(1) The synthesis and externalization of ADH and ALD decreased, at the same time,the level of ANP and AQP-2 increased.(2) The activities of SOD,Na⁺-k⁺-ATPase,Ca²⁺-Mg²⁺-ATPase and SDH in urinary detrusor strips are reduced.The frequency of spontaneous contraction was elevated significantly; The complaisance and elasticity of bladder were both attenuated.Compared with the young adult rats,significantly decreased relaxation response to high concentration of ISO and increased contractile response to ATP were found in aged model.Some histological changes were observed in aging model:the quantity of nephron was decreased,such as nephric tubule;glassy degeneration appeared in proximal convoluted tubule;the alinement of muscle fiber which in tunica muscularis vesicae urinariae was chaotic;fibrosis for vessel wall; the percentage of collagen fibers are increased.2,The pharmacologic action of SQWThe results show that SQW reduce significantly the urine and the concentration of sodium,chlorine in urine,and elevate the excretion of urine potassium in aging rats.It also can improve the frequency of urine and extend incubation period of urine.SQW can increase the releasing of ALD,ADH and the concentration of AQP-2 in urine,meanwhile,it can reduce the content of ANP.SQW can delay the proceeding of aging in detrusor muscle.The activity of SOD,Na⁺-k⁺-ATPase,Ca²⁺-Mg²⁺-ATPase,SDH and complaisance and elasticity of bladder increased after SQW administration in aging rats.The frequency of spontaneous contraction was cut down significantly.The decreased relaxation response to ISO and increased contractile response to ATP were also changed after SQW administration.The pathological characteristics of kidney and bladder function were switched:nephric tubule analosis was improved;the hyperplasia of collagen fibers which in tunica muscularis vesicae urinariae and thickening,chaotic muscle fiber was decreased. 3,The molecular pharmacology mechanism of SQWThe results showed that both mRNA and protein expressions of collagenâ… , collagenâ…¢,P2X₂ were decreased and mRNA and protein expressions ofβ₃-AR was increased in D-galactose mimetic aging model with polyuria afer SQW administration.SQW can recover the normal physiologic function of detrusor of bladder by myogenic and neurogenic ways.Conclusion:Sucessful duplication of subacute aging animal model with polyuria provided a reliable and effective model for our study.SQW reduced significantly the urine through effecting the production and excretion of urine;it can recover the normal physiologic function of detrusor of bladder. This recovery might be through up-regulatingβ3-AR mRNA and its protein expression and down-regulating collagenâ… ,collagenâ…¢,P2X2 mRNA and their protein expression.