Study Of The Effect And Mechanism Of Suo Quan Wan On Overactive Bladder In Diabetic Mice

ObjectiveEstablish an animal model of diabetes,observe the development of the bladder in the course of diabetes,and explore related molecular mechanisms.The effect of Suo Quanwan(SQW)on diabetic bladder was observed by intragastric administration,and explored its possible molecular mechanism.Methods1.Establishment and dynamic observation of diabetic bladder model(1)Establishment of animal modelsDiabetes models were established by STZ+ high-fat diet in C57BL/6J male mice.Normal group was fed normal feed,and model group for high-fat diet.Model group was injected streptozotocin(STZ)(100 mg)four times to model diabetic mice.After successful modeling,the mice were tested at 0W,3W,6W,9W and 12W.(2)Observation of general testsAfter successful animal modeling,the mice were tested for body weight,water intake,urine output,frequency of urination,fasting blood glucose and glucose tolerance at 0W,3W,6W,9W and 12W.(3)Observation of urinary dynamicThe urodynamic method dynamically measures the maximum bladder pressure,maximum bladder capacity,non-voiding contraction frequency,residual urine volume,urinary efficiency,and bladder compliance.(4)Observation pathologyThe changes of bladder structure were observed dynamically by HE andMASSON staining.(5)Observation of bladder detrusor functionThe in vitro tissue perfusion system dynamically detected the autonomic contraction of isolated bladder muscle strips and the effect of isolated muscle strips on ?,?-meATP,Carbachol,KC1,EFS-induced isolated muscle strips.(6)Changes in expression of Myosin Va and SLC17A9 in bladderThe mouse bladder was obtained,and the expression of Myosin Va and SLC17A9 protein in the bladder of mice was detected by western blot and RT-PCR.2.Observation of the effect of SQW on diabetic bladder(1)Observation of drug administration and general indicatorsAccording to the results of 1 to determine the modeling method and the progression of the disease to determine the time of occurrence of diabetes overactive bladder.The same method was used to mold.After successful modeling,it was divided into model group,positive group(tolterodine),SQW high-dose group,SQW medium-dose group,and SQW low-dose group.The model was successful 3W.At the beginning of the oral administration,when the model was successfully 6W,SQW were observed on the diabetic bladder.(2)Effect of SQW on detection of urodynamics indexUrodynamics was used to determine the effects of SQW on maximal bladder pressure,maximum bladder volume,number of non-urinary contractions,residual urine volume,urinary efficiency,and bladder compliance.(3)Effect of SQW on bladder structure The effects of SQW on bladder structure were observed by HE and MASSON staining.(4)Effect of SQW on detrusor function of bladder in vitro In vitro tissue perfusion system was used to observe the effect of shrinkage pills on bladder detrusor function.(5)Effect of SQW on the expression of Myosin Va and SLC17A9 Western blot and RT-PCR were used to observe the effect of SQW on its expression.Results1.Dynamic observation of diabetic bladderThe number of non-urination contractions in the model group increased first and then decreased,and 3-9W was significantly higher than the normal group,reaching a peak at 6W.The maximal pressure of the bladder in the model group increased first and then decreased,and was significantly lower than the normal group at 12W.Pathological results showed that the bladder weight and bladder weight to body weight ratio in the model group were significantly higher than those in the normal group at 3-12 weeks.The bladder wall thickness of the model group was significantly higher than that of the normal group and continued to increase;there was no significant difference in the smooth muscle-collagen ratio.The in vitro results showed that the model group showed a significant increase in the contraction amplitude of 3W and 6W compared with the normal group,and 12W significantly decreased.After adding ATP,the model group 6W,9W increased significantly compared with the normal group,and decreased significantly at 12W.When the model group was 6W,the concentration was significantly increased at 10-6M,3*10-6 M,and 1*10-5 M,and at 9 W,the concentration was 3*10-6 M,and the concentration of 1*10-5 M was significantly increased.At 12W,the concentration was 3*10-6 M,and 1*10-5 M was significantly decreased.Others showed no significant difference.In the model group,the contraction tension of the muscle strips at 16,32,and 64 Hz was significantly higher than that in the normal group;the contraction tension of the muscle strips at 8,16,32,and 64 Hz was significantly higher in the 9W model group than in the normal group;The contraction tension of the muscle strips at 8,16,32,and 64 Hz was significantly lower than that of the normal group.Myosin Va protein and mRNA levels were significantly higher in the model group than in the normal group at 6W,and significantly lower than the normal group at 12W.The model group SLC17A9 was significantly higher than the normal group at 6-12W,and its mRNA expression was significantly higher than that of the normal group at 6W and 9W.2.The effect of Suo Quan Wan(SQW)on diabetic bladderCompared with the normal group,the weight and urination efficiency of the model group were significantly reduced.The model group received 24 hours of drinking water,fasting blood glucose,glucose tolerance,area under the curve,non-urinary contraction frequency,maximum bladder volume,residual urine volume,and bladder compliance were significantly increased.Compared with the model group,the number of non-urinary contractions in the high-,medium-,and low-dose SQW groups and the positive group were significantly reduced;the maximum bladder capacity in the middle group of SQW was significantly reduced;the residual urine in the positive group and medium SQW groups were significantly reduced;the bladder compliance in medium SQW group was significantly decreased;the urinary efficiency of the positive group and was significantly increased.Compared with the normal group,the bladder muscle strip contraction response in model group was significantly increased when given ATP,32 Hz,64 Hz,10-7-10-5 M.Compared with the model group,the contraction response of muscle strips was significantly reduced in the positive group and SQW high,medium dose groups after adding ATP;the contraction response was significantly reduced after adding KC1 in the positive group and medium SQW dose group;32Hz,64Hz stimulation,the middle SQW group was significantly reduced;the contraction of the positive was significantly reduced at 10-5 M,and the contraction response of the muscle strips in the low-dose group of the SQW was significantly reduced at the concentration of 10-7-10-5 M.Compared with the normal group,the expression of Myosin Va and SLC17A9 protein and mRNA in the model group were significantly increased.Compared with the model group,the expression and expression of Myosin Va protein in the group were significantly decreased.The SMC17A9 in the dose group of the shrinking spring pill was significantly reduced.conclusionSQW can significantly improve the excessive activity of diabetes,which is related to the regulation of Myosin Va and SLC17A9.The mechanism may be to reduce the contraction of Myosin Va and SLC17A9 in the bladder to reduce the contraction of smooth muscle caused by the release of ATP,thereby improving the unstable contraction of diabetic overactive period and improving its symptoms.