The Effects Of Endothelin And Nitrogen Monoxidum On Neurogenic Baldder

PART ONETHE CLINICAL SIGNICANCE OF ENDOTHELIN AND NITROGEN MONOXIDUM DETECTED IN PLASMAAND URINARYObjective: To explore the quantity of endothelin and nitrogen monoxidum in blood plasm and urinary and it's significance in neurogenic bladder diseases.Methods: The 22 patients of neurogenic bladder were from the period during 2003.10²004.8, 10 months¹4 years old, the average was 6.28±3.12 years old. All patients went gone urodynamic study, nervous electromyogram, voiding cysto urethro graphy, intravenous pyelography(IVP). Using radio-immunity, we detected the contents of endothelin and nitrogen monoxidum blood of empty stomach and urina sanguinis collected in the morning. And using chrome disoxidation biochemistry, we detected nitrogen monoxidum from blood and urinary, and used control group age match 10 cases (oblique inguinal hernia and funicular hydrocele). Using color Doppler, we detect renal perfusion speeds and renal impedance (RI), and calculate renal blood perfusion, using control group. We use computing ellipsoid cubic method to calculate renal cubic( 4/3×π×d₁/2×d₂/2×d₃/2)(d₁ ,d₂,d3,three diamensions diameter of kidney), and calculate renal cubic of unit volume and unit time, and calculate average urinary volume after keeping 3 days urinary, and measure blood pressure.Results: all patients suffered to hyperreflexia. The endothelin-1 in plasma was 0.478±0.039 fmol/ml, and in the urinary was 0.453±0.029 fmol/ml in neurogenic bladder. Nitrogen monoxidum in plasma was 69.57±36.68μmol/, and in the urinary was 392.36±121.1 μmol/L in neurogenic bladder. The endothelin-1 in plasma was 0.478±0.039 fmol/ml, and in the urinary was 0.548±0.073 fmol/ml in normal children. Nitrogenmonoxidum in plasma was 74.56±11.39 umol/, and in the urinary was 268.92±88.38 umol/L in normal children. Renal cubic of unit volume and unit time was 8.37±2.26ml/min.cm3, and renal impedance ( RI) was 0.667±0.049, and average urinary was 1000±156ml/24h, and blood pressure was 85⁹5/60⁷5mmHg. Endothein-1 in plasma was higher than in urinary(p<0.05), and NO in the urinary was higher than that of normal children(p<0.05). Endothelin-1 and nitrogen monoxidum of plasma in neurogenic bladders were not significant with normal children(p>0.05). Renal blood perfusion had no relationship to renal impedance and vesical pressure of filling of bladder, also had no relationship to the content of endothelin-1 in plasma.Conclusions: Endothelin and nitrogen monoxidum in plasma were at the normal level in hyperreflexia patients, but in the urinary lied disproportion of endothelin and nitrogen monoxidum, and endothelin was higher, and nitrogen monoxidum was lower than normal. Mucosal barrier functional defect in bladder and urethra lied in neurogenic bladder patients, and the contents of urinary may interdiffused with tissues of bladder wall. The less endothelin in urinary occurred when endothelin of urinary perfused bladder tissues, of tissues produced even more less endothelin, or endothelin was used more in bladder tissues. Mechanism of nitrogen monoxidum augmented externalization may occurred in urinary system, and the theory of effects of augmented nitrogen monoxidum on urinary passage which were passive or positive, and the relationship with endothelin which were opposition or antagonism was not determined. Endothelin and nitrogen monoxidum in plasma were at the normal level in hyperreflexia patients, and had no effects on haemodynamics all over the body, but it's effects on kidney and bladder can't be confirmed using simple clinic examination methods.PART TWOTHE BIOLOGICAL SIGNICANCE OF EXPRESSION OFENDOTHELIN AND IT'S RECEPTORS INNEUROGENIC BLADDER TISSUESObjective :to explore the expression of endothelin-123 ( ET-123), endothelin receptor A (ETA) and endothelin receptor B(ETB) in neurogenic bladder tissues, and analyze it's producing feature and significance.Method: there were 30 cases with neurogenic bladder, and malecases, and 12 female cases. The average 6.1±1.91 years old. All patients appeared with myelodysplasia: tethered cord syndrome, cleftspine, meningomyelocele. 22 cases with caudex dorsalis lysis, and 8 cases with no surgical treatment. All patients suffered from urinar y and fecal incontinence in different degree. All patients went goneurodynamic study, nervous electromyogram, voiding cysto urethro g raphy, intravenous pyelography(IVP). All patients were manifested w ith hyperreflexia bladders, and treated with surgical procedures. Thesurgical procedure were suspension of pelvic floor, suspension of b ladder neck, bladder augmentation, Malone procedure, and anti-ureter al reflux. During operation, collected bladder tissue samples includin g tissues of apex vesicae and tissues of bladder neck, and all tissue s were enveloped with mineral wax. All tissues were detected for e ndothelin-123, endothelin receptor A and endothelin receptor B respe ctively in tissues of apex vesicae and tissues of bladder neck, and with normal bladder tissues as control group (bladder tissues of hyp ospadia, 10 cases), and according to clinical features, to explore theexpression of endothelin-123, endothelin receptor A and endothelin receptor B, and to analyze the relationship among them.Results: there were 13 cases complicatin nephrydrosis, with lateral nephrydrosis 3 cases and bilateral 10 cases, bilateral ureteral ref lux 10 cases, and 2 cases with lateral ureteral reflux, with hyperrefl exia 30 cases, outlet obstruction 22 cases, detrusor of bladder and musculus sphincter urethrae membranaceae mis- match 22 cases, andmatch 5 cases. Nervous electromyogram showed muscle of pelvic fl oor in denervation and myoparalysis. Nervous lesion involved in late ral extremitas inferior or bilateral extremitas inferior, and showing t alipes cavus, claudication, decreasing of muscles of lower limb. 28 cases with small bladder in voiding cysto urethro graphy, and irregu lar bladder, and diverticula, and bladder neck below articulation of pubis. The surgical procedures suspension of pelvic floor, suspensionof bladder neck, bladder augmentation, Malone procedure, and anti-ureteral reflux were used in those patients, with 4 types of surgicalprocedures 10 cases(33.3%), and 3 types of that 17 cases(56.6%), 2 types of that 3 cases(10%), and the arverage is 3.23 (97/30) kind s of surgical procedures. There were 30 cases apex vesicae tissues a nd 10 cases bladder neck tissues, and normal apex vesicae tissues 1 0 cases, and normal bladder neck tissues 10 cases. In normal bladde r tissues, smooth mscules were stained with endothelin-123 ( ET-12 3), endothelin receptor A (ETA) and endothelin receptor B(ETB), an d there was no significance between endothelin receptor A and endo thelin-123(p>0.05), and much more than endothelin receptor B, and endothelin receptor B distributed in bladder tissues had no differenc e between tissues of apex vesicae and tissues of bladder neck..Therewere less interstitial tissue of urinary bladder, and there had no en dothelin receptor A and endothelin-123 expression, or light stained i n collagen fibers( CF) and collagenoblast, and had no expression ofendothelin receptor B there. In neurogenic bladder tissues, much m ore interstitial tissue of urinary bladder over there, there were muchmore collagen fibers( CF) and collagenoblast, and the expression ofendothelin receptor A and endothelin receptor B higher than normal tissues(p<0.05), especially in bladder neck tissues, the most expres sion of endothelin receptor A occurred(p<0.05), and endothelin recep tor B had no expression in interstitial tissues. The expression of en dothelin-123 ( ET-123) in smooth muscles are stained, and less expr ession in tissues of apex vesicae than normal, and more expression in tissues of bladder neck, and there was no or less expression in i nterstitial tissues.Conclusions: In normal bladder tissues, endothelin-123 may adjust vesical tensile force through combining with endothelin receptor A (ETA) and endothelin receptor B(ETB), and smooth muscle cells can produce endothelin, and smooth muscle cells have characteristic of autocrine and paracrine, and the main action was caused by endothelin A, not only maintain vesical tensile force, but also may sustain the quantity and composition of bladder base materials, and endothelin receptor A may hold normal urethra pressure in bladder neck, and endothelin receptor B may only sustain vesical tensile force and had no effects on base materials. In neurogenic bladder, much more endothelin receptor A expressed than in normal tissues, and may play very important role in smooth muscles proliferation, and in the bladder neck tissues may play important part in bladder outlet, and in the base material may promote fibroblastic proliferation and collagen formation. The expression of endothelin receptor B may hold vesical tensile force of smooth muscle, and had no effects on base materials. The decrease of endothelin-123 in tissues of apex vesicae may related to the quantity of smooth muscle cells or apomorphosis of cells, or less blood supply.PART THREETHE BIOLOGICAL SIGNIFICANCE OF EXPRESSION OF NITRIC OXIDE SYNTHASES IN NEUROGENICBLDDER TISSUESObjective :to explore the expression of nitric oxide synthases including neuronal nitric oxide synthases(nNOS), inducible nitric oxide synthases(iNOS), endothelial nitric oxide synthases (eNOS) in neurogenic bladder tissues, and analyze it's producing feature and significance.Method: there were 30 cases with neurogenic bladder, and malecases, and 12 female cases. The average 6.1±1.91 years old. All patients appeared with myelodysplasia: tethered cord syndrome, cleftspine, meningomyelocele. 22 cases with caudex dorsalis lysis, and 8 cases with no surgical treatment. All patients suffered from urinar y and fecal incontinence in different degree. All patients went goneurodynamic study, nervous electromyogram, voiding cysto urethro g raphy, intravenous pyelography(IVP). All patients were manifested w ith hyperreflexia bladders, and treated with surgical procedures. Thesurgical procedure were suspension of pelvic floor, suspension of b ladder neck, bladder augmentation, Malone procedure, and anti-ureter al reflux. During operation, collected bladder tissue samples includin g tissues of apex vesicae and tissues of bladder neck, and all tissue s were enveloped with mineral wax. All tissues were detected for n euronal nitric oxide synthases, inducible nitric oxide synthases and e ndothelial nitric oxide synthases respectively in tissues of apex vesi cae and tissues of bladder neck, and with normal bladder tissues ascontrol group (bladder tissues of hypospadia, 10 cases), and accord ing to clinical features, to explore the expression of neuronal nitric oxide synthases, inducible nitric oxide synthases and endothelial nitric oxide synthases, and to analyze the relationship among them.Results: there were 13 cases complicatin nephrydrosis, with late ral nephrydrosis 3 cases and bilateral 10 cases, bilateral ureteral ref lux 10 cases, and 2 cases with lateral ureteral reflux, with hyperrefl exia 30 cases, outlet obstruction 22 cases, detrusor of bladder and musculus sphincter urethrae membranaceae mis- match 22 cases, andmatch 5 cases. Nervous electromyogram showed muscle of pelvic fl oor in denervation and myoparalysis. Nervous lesion involved in late ral extremitas inferior or bilateral extremitas inferior, and showing t alipes cavus, claudication, decreasing of muscles of lower limb. 28 cases with small bladder in voiding cysto urethro graphy, and irregu lar bladder, and diverticula, and bladder neck below articulation of pubis. The surgical procedures suspension of pelvic floor, suspensionof bladder neck, bladder augmentation, Malone procedure, and anti-ureteral reflux were used in those patients, with 4 types of surgicalprocedures 10 cases(33.3%), and 3 types of that 17 cases(56.6%), 2 types of that 3 cases(10%), and the arverage is 3.23 (97/30) kind s of surgical procedures. There were 30 cases apex vesicae tissues a nd 10 cases bladder neck tissues, and normal apex vesicae tissues 1 0 cases, and normal bladder neck tissues 10 cases. In normal apexvesicae tissues, all cases stained with neuronal nitric oxide synthas es(nNOS), and distributed among bundles of smooth muscles, and su rface of smooth muscles and interstitial tissue, histochemical score( HS) 2.8-4.0 and 1.2-2.7. There were no stained cells in bladder tis sues of inducible nitric oxide synthases(iNOS), and histochemical sc ore( HS) was very lower, HS: 0-0.4 and 0-0.1; Endothelial nitric o xide synthases (eNOS) mainly distributed in interstitial tissues in rar efaction manners, and mainly in vascular endothelial cell (VEC), an d smooth muscles had no staining; the most expression among themwas nNOS, and mainly distributed in bladder neck tissues. In neurogenic bladder tissues, the main expression of NOS type was iNOS, and nNOS decreased significantly, the trend of NOS type was iNO S>nNOS>eNOS. And eNOS mainly expressed in vascular endothelial cell (VEC) among interstitial tissues, and had no staining in smoot h muscle cells and collagenoblast and rarefaction of microvessel in bladder tissues, and microvessel density decreased significantly than normal bladder tissues. Microvessal density(MVD) in bladder tisssus (6.8±3.2/100 per square) was lesser than that in normal tissues (616. 7±6.28/100 per square).Conclusions: In normal bladder tissues, neuronal nitric oxide synthases(nNOS) mainly distributed in bladder neck and urethra, and nitric oxide mainly derived from nNOS. Much more matrix fibers was seen in neurogenic bladder interstitial tissue, and less nitrogenergic nerve, and less nNOS expression. Those change may have important role in outlet obstruction of bladder. There were much more inducible nitric oxide synthases(iNOS) expression in bladder tissues, and NO mainly derived from iNOS, and it may play important in pathological bladder tissues, especially in fibrosis of bladder wall. NO from iNOS may block P-adrenergic receptor, inhibit atony of bladder, and affect bladder storing. Endothelial nitric oxide synthases (eNOS) may be considered as angiopoietic labeling, and may evaluate blood supply of bladder. The tissues in bladder had less blood supply than that in normal bladder tissues.