Effect And Mechanism Of Electroacupuncture On Diabetic Neurogenic Bladder

Background: Diabetic neurogenic bladder(DNB)is one of the common chronic complications of type 2 diabetes mellitus(T2DM).As a characteristic therapy of traditional Chinese medicine(TCM),acupuncture has been proven effective in improving urinary system diseases and neurological dysfunction.Because the pathogenesis of DNB has not been fully elucidated and the factors affecting its prognosis are complex,despite the multitude of modern medical methods for DNB treatment,the efficacy is not satisfactory.As a combination of traditional acupuncture and modern technology,EA has become a commonly used clinical treatment method in TCM.EA has advantages in the treatment of diabetic neuropathy,however,its clinical efficacy and mechanism of action in patients with DNB are still uncertain.Objective: The clinical trials were conducted to observe the therapeutic effect of EA on patients with DNB,evaluate the effectiveness of EA in improving patients’ urinary system symptoms and quality of life,and explore the safety and clinical promotion value of EA in the treatment of DNB.To further observe the effect of EA on the bladder function of streptozotocin-high-fat diet induced DNB model rats,and to explore the mechanism of EA on the phosphorylation level of myosin system in the PACAP/c AMP/PKA signal transduction pathway of bladder detrusor in type 2 DNB model rats.Methods:(1)Sixty participants who met the criteria were randomly divided into the control group(n=30)and EA group(n=30).Both groups were given individualized basic interventions,including healthy lifestyle education,hypoglycemic treatment,neurotrophic treatment,and water intake planning.The control group only received basic intervention measures,and the EA group was treated with EA therapy in addition to basic intervention.Shenshu,Pangguangshu,Zhongliao,and Sanyinjiao were treated with EA(density wave,10/50 Hz).EA was performed for 30 min each time,5 times a week,for 8 consecutive weeks.The clinical efficacy of EA for DNB was evaluated by observing the changes in the TCM syndrome score,bladder residual urine volume,maximum urine flow rate,AUA-SI score,SF-Qualiveen score,and glucose metabolism-related indexes in the two groups before and after treatment.The adverse events during the trial were recorded and statistically analyzed.(2)A total of 50 female rats were randomly divided into the normal group(n =10)and model group(n = 40).The normal group was fed with an ordinary diet,and the model group was fed with a high-fat diet combined with a single intraperitoneal injection of STZ 35 mg/kg to induce type 2 DNB.After 8 weeks,blood glucose was measured,and the moisture of the padding in the feeding cage was observed to evaluate whether the DNB rat model was successfully constructed.The DNB model rats were randomly divided into 3 groups(EA group,sham EA group and the model group),with 10 rats in each group.In the EA group,EA(density wave,10/50 Hz)at Shenshu,Pangguangshu,Zhongliao,and Sanyinjiao points was administered for 30 min each time,5 times a week,for consecutive 8 weeks.In the sham EA group,the acupuncture needles were superficially punctured and fixed at the same position on the body surface of the EA group.However,the acupuncture needles did not penetrate the acupoints,and the electrostimulation apparatus was turned off.After the intervention,the body mass of all rats was measured,and the 24-h urine volume was recorded by the metabolic cage.An Intraperitoneal glucose tolerance test was performed in each group.The insulin levels in each group were detected by ELISA.Urodynamics was used to detect the leakage point pressure,maximum bladder capacity,bladder base pressure,and bladder compliance of all rats.Bladder wall thickness was measured by ultrasonography.After the bladder wet weight test,histopathological changes in the bladder detrusor were observed by hematoxylin–eosin(HE)staining.Ultrastructural changes in the bladder detrusor of each group were observed by transmission electron microscopy(TEM).The expressions of PACAP38,c AMP,and PKA in the bladder detrusor were detected by ELISA.RT-PCR was used to detect the PACAP38 m RNA expression in the bladder detrusor muscle of all rats.The PACAP38,p-MLCK,p-MYPT1,and p-MLC expression in the bladder detrusor muscle were detected by Western blot.Results:(1)After treatment,the levels of Hb A1 c,FBG and 2h PBG in the two groups were significantly lower than those before intervention(P < 0.05).However,no significant difference was found between the two groups.After treatment,the residual urine volume of the bladder in the two groups decreased significantly(P <0.05).The improvement in the residual urine volume of the EA group was better than that of the control group(P < 0.05).The maximum urinary flow rate of the two groups was higher than that before treatment(P < 0.05).The improvement degree of maximum urine flow rate in the EA group was better than that in the control group(P< 0.05).The TCM syndrome score of both groups decreased after treatment compared with that before treatment,and the score of the EA group was significantly lower than that of the control group(P < 0.05).After treatment,the AUA-SI score in the EA group decreased,and the decreasing range was higher than that in the control group(P< 0.05).The SF-Qualiveen score in the EA group decreased after treatment,and the change range in the SF-Qualiveen score of the EA group was higher than that of the control group(P < 0.05).No adverse events were observed during the trial.(2)Compared with the normal group,the serum INS,HOMA-IR,24-h urine volume,and bladder wet weight in the model group significantly increased,and the body mass significantly decreased(P < 0.05).The urodynamic test showed that the maximum bladder capacity and bladder compliance of the model group were significantly increased(P < 0.05)and the leak point pressure decreased(P < 0.05)compared with those in the normal group.Ultrasonic examination showed that the bladder wall thickness of the model group was significantly higher than that of the normal group(P < 0.05).After 8 weeks of EA treatment,the wet bladder weight of the EA group was significantly lower than that of the sham EA group.In the EA group,the bladder wall thickness decreased,the maximum bladder capacity and bladder compliance decreased,and the leakage point pressure increased(P < 0.05).No significant difference was found between the sham EA and model groups.HE staining showed that EA could reduce the inflammatory cell infiltration and relieve hyperemia and edema in the bladder detrusor muscle of type 2 DNB model rats.The TEM test showed that EA could repair the damaged mitochondria of bladder detrusor cells.The ELISA showed that the levels of PACAP38,c AMP,and PKA in the bladder detrusor of the model group were significantly lower than those of the normal group(P < 0.01).EA can upregulate the c AMP and PKA levels in DNB model rats(P < 0.05).RT-PCR results revealed that the bladder detrusor PACAP38 m RNA expression in the model group was significantly lower than that in the normal group(P < 0.01),and EA treatment could upregulate the PACAP38 m RNA expression in the type 2 DNB rat model(P < 0.05).Western blot results showed that compared with the normal group,the levels of PACAP38 and p-MLCK in the bladder detrusor of the model group were significantly decreased,and the p-MLC level was significantly increased(P < 0.01).EA treatment could significantly upregulate PACAP38 and p-MLCK levels in type 2DNB rat models(P < 0.01)and downregulate p-MLC levels(P < 0.05).Conclusions:(1)The clinical trials have demonstrated that EA can empty the bladder by reducing bladder residual urine volume and increasing the maximum urine flow rate in patients with DNB.EA can improve TCM syndrome and AUA-SI scores in patients with DNB,relieve urinary symptoms,and improve bladder function.EA can improve the quality of life of patients with DNB,and its curative effect is accurate and safe.(2)Animal experiments demonstrated that EA can activate the PACAP/c AMP/PKA signal transduction pathway in the DNB model rat detrusor to regulate the phosphorylation level of myosin system-related contraction elements.EA can repair the damaged mitochondria of detrusor cells to improve energy metabolism,thus relieving the pathological degree of inflammatory cell infiltration,hyperemia,and edema of the bladder detrusor.EA can reverse bladder remodeling to improve bladder hypertrophy,regulate bladder pressure and compliance,and repair the damaged bladder function of type 2 DNB model rats.