Basic Research Of Transcutaneous ENS And Coffee Or Caffeine On Bladder Dysfunction In Diabetic Cystopathy

Diabetes mellitus is a common endocrine and metabolic disease. The diabetic neuropathy is one of the most common chronic complications of the diabetes. Diabetic cystopathy (DCP) is one of diabetic peripheral nerve pathological changes, belonging to neuropathic dysfunction of bladder (NB).Being one of the complications of urinary system caused by diabetes, its incidence occupies about 50% of all diabetic patients. Clinical researches illustrate that even if those whose blood glucose is controlled steadily, the incidence of DCP is still 25%.The definite pathogenesis of DCP is not completely known until now. Present research considers that the pathogenesis principally concludes two aspects: peripheral and autonomic nerve pathological changes, myopathic abnormal factors and so on.To seek effective methods, to prevent actively as early as possible, and to treat diabetic cystopathy pathological changes, to improve lower urinary tract symptom are important components for diabetic patients combined treatment and to raise quality of life. At present therapy of diabetic cystopathy is mainly drug therapy. Drug therapy has some effect, but side effect should not be ignored. Invasive treatment contains intermittent clean catheterization, cystostomy, transurethral resection bladder neck, and they can relieve much residual urine. Surgical invasive treatment can relieve urinary tract infection and upper urinary tract damage, but it cannot resolve fundamental problem. It may cause further infection if usage is incorrectness. How to find an effective and safe method to prevent and treat DCP pathological changes, to relieve detrusor lacking strength and much residual urine is a problem people have being explored. To prevent the DCP develops in early stage is the key to combined modality therapy and improve quality of life to diabetic patients.The reports of using Transcutaneous Electrical Nerve Stimulation(TENS) are few, and there are no researches for urodynamics and pathogenesis. Another research discovers that frequent drinking coffee can decrease the incidence of 2-diabetes.The effect of coffee for DCP is still unknown.This research is to investigate the effects of TENS on the urinary bladder voiding dysfunction in diabetic cystopathy rats by SD rat model of diabetic cystopathy rat was established. And using coffee or caffeine to perfuse diabetic cystopathy rat bladder with nasogastric tube caffeine aqueous solution(10mg·kg-1·d-1)and Nestle coffee aqueous solution. Through urodynamics, cytobiology, biochemistry and molecular biology, investigating pathogenesis and effect that TENS and coffee or caffeine have on DCP in neuropathic and myopathic aspects. It is an simple, convenient and economical way for diabetic patients to apply TENS technology and drink proper coffee or caffeine every day. It would have a great significance in economic benefit and social benefit.PartⅠⅠ:Urodynamics and pathology study in diabetic cystopathy rat Objective:To observe the abnormalities of urodynamics and histopathology in the early stage of DCP. To find a relationship between the changes in the bladder function and detrusor structure and study the onset mechanism and pathology procedure in order to provide scientific bases for early stage diagnosis and treatment.Materials and Method:66 Sprague-Dawley male rats were randomly divided into 2 groups:NC group (normal control, n=30), DM group (diabetes mellitus, n=36),. Diabetic rat model was induced by injection of streptozotocin(STZ)into peritoneal cavity. Diabetic rats were successfully accepted as diabetes by measuring fasting serum glucose concentration more than 12mmol·L-1 continually three times. In both group: the bladder resting pressure, the bladder maximum capacity, the maximum bladder pressure and bladder compliance were performed. The bladder weights were taken, bladder wall thicknesses were measured and the histopathologic structures of bladder walls were observed by optical and electric microscopy.Results: In the diabetic rats, the the maximum bladder capacity (P<0.01), compliance (P<0.05) and resting pressure (P<0.01) were increased, however the maximum bladder pressure decreased compared to the control group (P<0.01). The weight of diabetic rat bladder was increased compared to the control bladders (P<0.05). In the diabetic rats, not only the total bladder wall thickness was greater but also the muscle bundles appeared to be increased in size. In contrast to the control group, the diabetes rats showed in the detrusor cell obvious karyopyknosis, a number of compact bodies decreased and cell junction fibrosis between the detrusor cells, fibroblast degeneration and collagen fibers array disorders.Conclusions: In this study, the urodynamics in the diabetic bladders showed significant differences compared to the normal bladders. The detrusor microstructure showed to be abnormal in the early stage of diabetes. It is suggested that the bladder function is impaired obviously in the early stage of diabetes. The result of the study can be helpfully to fumble diabetic progress of bladder dysfunction in patient and therapeutic results.PartⅡ:Effects of TENS on urinary blaader voiding dysfunction in diabetic cystopathy ratsObjective:To investigate the effects of Transcutaneous Electrical Nerve Stimulation (TENS) on the urinary bladder voiding dysfunction in diabetic cystopathy rats.Methods:SD rat model of diabetic cystopathy rat was established by the injection of STZ in abdominal cavity. The rats were randomized into the DM ES group(DM with electrical stimulation,n=18), DM(diabetes mellitus,n=18) group and normal control group(n=14). TENS was performed on rats in the DM ES group in the 10th week after the establishment of diabetic rats model (stimulation parameter: intensity--31V, density—31Hz;two pairs of electrode was place on the projective area on the abdomen of the bladder respectively) , but those in the control group underwent shamoperation . After the stimulation was continuing for 3 weeks, The bladder wet weight, volume threshold for micturition, post-voided residual volume, bladder residual volume/bladder capacity (R%),the max micturition pressure, the changes of the detrusor strip contractility and bladder muscle ultrastructure were detected . The expression of CGRP mRNA in each rats'DRG was measured by RT-PCR,The expression levels of CGRP of Bladder and DRG were detected by western blot.Results: Compared with DM group , the R% and volume threshold for micturation were significantly reduced while the contractility of the detrusor muscle strip were significantly increased in DM ES group after electrical stimulation. Histopathological studies showed that the detrusor cells expressed compensatively hypertrophy, intracellular collagen fibers increased obviously and mitochondra swelled in the bladder muscle unltrastructure of diabetic rats. Electrical stimulation could reduced markedly the compensated hypertrophic cells, intercellular collagen gibers and protect mitochondria in the bladder muscle ultrastructure of DM ES group, The content of the mRNA of CGRP in DRG of DM ES group is enhanced than that in DM group. The expression levels of CGRP of Bladder and DRG in DM ES group is increased than that in DM group.Conclusion: These experiment results demonstrate that the TENS can promote the voiding in the DCP(diabetic cystopathy)rats, TENS may enhance the expression levels of CGRP of DRG and bladder in DCP, Which react in the bladder through the effective conduction, improve the bladder fullness, restore the initiate factor of the micturition. Otherwise TENS also ameliorate the configuration and function of ultrastructure, regulate the intracellular Ca2+ concentration to facilitate bladder contractility. TENS maybe accelerate the restoration of the nerver through enhace. The expression levels of NGF or the injury/regeneration related gene.PartⅢ:Effects of coffee or caffeine on DCP ratsObjective: To observe histomorphological abnormality and functional alterations of diabetic cystopathy in the streptozotocin-induced diabetic rat and explore the effects and mechanisms of caffeine and coffee on the diabetic bladder dysfunction Materials and Methods: 75 Sprague-Dawley male rats were randomly divided into 4 groups: NC group (normal control, n=15), DM group (diabetes mellitus, n=20), Coffee group (DM with coffee treatment, n=20) and Caffeine group (DM with caffeine treatment,n=20). Diabetic rat model was induced by injection of streptozotocin(STZ)into peritoneal cavity. Diabetic rats were successfully accepted as diabetes by measuring fasting serum glucose concentration more than 12mmol·L-1 continually three times. Caffeine dissolved in distilled water were administered orally (10mg/kg/day, p.o.) to diabetic rats of caffeine group for 7 weeks. According to this concentration, diabetic rats of coffee group received corresponding coffee treatment by the same method. The rest two groups were given orally the same volume of distilled water every day. After 7 weeks, the bladder wet weight,volume threshold for micturation,post-voided residual volume (PVR), V%(voided volume/bladder capacity),cAMP content of bladder, the changes of the detrusor strip contractility and bladder muscle ultrastructure were measured and compared among the groups by the methods of urodynamics, detrusor strip contractile studies, radioimmunoassay and histopathology.Results: After 7 weeks, the bladder wet weight, volume threshold for micturition and PVR were significantly higher in the diabetic rats of DM group than those in NC group(P<0.01), while V% and cAMP content of bladder were significantly lower in DM group(P<0.01). Coffee or caffeine treatment significantly reduced the bladder wet weight, bladder capacity and PVR as well as increased V% and cAMP content of bladder in the diabetic rats(P<0.05). In addition, coffee and caffeine tended to normalize the altered detrusor contractile responses to electrical field stimulation and acetylcholine in the diabetic rats. Histopathological studies showed that the detrusor cells expressed compensatively hypertrophy, intracellular sarcoplasmic reticulum and intercellular collagen fibers increased obviously and mitochondria swelled in the bladder muscle ultrastructure of diabetic rats. Coffee or caffeine treatments reduced significantly the compensated hypertrophic cells, intracellular sarcoplasmic reticulum and intercellular collagen fibers and protect mitochondria in the bladder muscle ultrastructure of diabetic rats.Conclusions: These results indicate that the bladder functions were impaired obviously in the early stage of diabetic rats. Caffeine and coffee may have beneficial effects on bladder dysfunction in the early stage of diabetic rats by up-regulation of cAMP content in the lower urinary tract mediating cell-signal transduction pathways, improvement of the bladder fullness sensation, recovery of the micturition reflex and protection of bladder ultrastructure regulating intracellular Ca2+ concentration to potentiate bladder contractility.