Exploring The Therapeutic Effect Of Methylcobalamin Ciliao Acupoint Injection Therapy On Neurogenic Bladder In Rats With Suprasacral Spinal Cord Injury Based On The SCF/C-kit Pathway

ObjectiveObserving the morphological changes of bladder and kidney tissue,as well as the protein and m RNA changes of SCF,C-kit,JAK2,STAT3,M2 in bladder tissue of rats with neurogenic bladder after suprasacral spinal cord injury treated with methylcobalamin Ciliao point injection,and exploring the possible mechanism of methylcobalamin Ciliao point injection in treating neurogenic bladder rats after suprasacral spinal cord injury.MethodsPurchase 36 SPF grade SD female rats for 6-8 weeks,weighing(220 ± 10)g.After one week of normal feeding,they were randomly divided into three groups:Sham surgery group,model group and acupoint injection group,with 12 rats in each group.The rats in the Sham surgery group only chewed out the vertebral lamina,and the rats in the model group and the acupoint injection group made T10 spinal cord injury models through the improved Allen’s method.After the rats had passed through the spinal cord shock period,the Sham surgery group and the model group received routine nursing care,without drug treatment.The acupoint injection group directly punctured about 15 mm at Ciliao,injected 0.1m L of mecobalamin every time,once a day,alternately,for 28 days.Each group of rats underwent manual urination daily and recorded their urination volume,3 times per day.After 28 days of treatment,specimens were euthanized for bladder weighing and bladder tissue collection for testing.Calculate the ratio of bladder weight to body weight in each group of rats,observe the pathological changes of bladder and kidney tissue with H&E staining,detect the protein expression of SCF,C-kit,JAK2,STAT3,M2 in bladder tissue with immunohistochemical method,detect the protein expression of SCF,C-kit,JAK2,STAT3,M2 in bladder tissue with Western blot,and detect the m RNA expression of SCF,C-kit,JAK2,STAT3,M2 in bladder tissue with RT q PCR method.Results1.Daily average manual urination: the daily average manual urination of rats in the model group and the acupoint injection group was higher than that in the sham operation group(P<0.05),and the daily average manual urination of rats in the acupoint injection group was lower than that in the model group(P<0.05),the difference was statistically significant2.Changes in body weight and bladder index of rats in each group: After treatment,the body weight of rats in the model group and acupoint injection group was significantly lower than that in the sham operation group(P<0.01),and the difference was statistically significant.There was no significant difference in body weight between model group and acupoint injection group.The bladder index of rats in the model group and acupoint injection group was higher than that in the sham operation group(P<0.05),and the bladder index of rats in the acupoint injection group was lower than that in the model group(P<0.05),the difference was statistically significant.3.H&E staining was used to observe the morphological changes of the bladder: in the sham operation group,the epithelial cells in the mucosa layer were arranged regularly,the cell structure and morphology were normal,the lamina propria contained more elastic fibers,and the muscular layer was thick;In model group,compared with sham operation group,more epithelial cells degenerated,cytoplasm vacuolized,more capillary congestion and dilation in lamina propria,accompanied by lymphocyte and neutrophil infiltration,disordered arrangement of muscle cells,and widened gap were seen in mucosa layer;Point injection group: compared with the model group,epithelial cell degeneration and cytoplasmic vacuolation can be seen locally in the mucosa layer,mild neutrophil and punctate lymphocyte infiltration can be seen in the lamina propria,and slight hypertrophy of muscle cells can be seen.4.H&E staining was used to observe the morphological changes of kidney: in the sham operation group,the cortical medulla was clearly demarcated,the glomerulus in the cortex was evenly distributed,the number of cells and matrix in the glomerulus were uniform,the epithelial cells of renal tubules were round and plump,the brush border was orderly arranged,there was no obvious abnormality in the medulla,there was no obvious hyperplasia in the interstitial,and there was no obvious inflammatory change;Model group: compared with sham operation group,multiple renal tubular atrophy,accompanied by connective tissue hyperplasia and a large number of lymphocyte infiltration can be seen in the renal cortex.Point injection group:compared with the model group,the degree of renal cortex atrophy decreased,the connective tissue slightly proliferated,and a small amount of lymphocytes infiltrated.5.The expression of SCF,C-kit,JAK2,STAT3,M2 protein in bladder tissue of rats in each group was detected by immunohistochemistry: the expression of SCF,C-kit,JAK2,STAT3,M2 protein in model group and acupoint injection group was higher than that in sham operation group(P<0.05),and the expression of SCF,C-kit,JAK2,STAT3,M2 protein in acupoint injection group was lower than that in model group(P<0.05),with statistical significance.6.The expression of SCF,C-kit,JAK2,STAT3 and M2 m RNA in bladder tissue of rats in each group was detected by reverse transcription quantitative polymerase chain reaction: the expression of SCF,C-kit,JAK2,STAT3 and M2 m RNA in bladder of model group and acupoint injection group was higher than that of sham operation group(P<0.05),and the expression of SCF,C-kit,JAK2,STAT3 and M2 m RNA in acupoint injection group was lower than that of model group(P<0.05),with statistical significance.7.The expression of SCF,C-kit,JAK2,STAT3,M2 protein in bladder tissue of rats in each group was detected by Western blot: the expression of SCF,C-kit,JAK2,STAT3,M2 protein in bladder of rats in model group and acupoint injection group was higher than that in sham operation group(P<0.05),and the expression of SCF,C-kit,JAK2,STAT3,M2 protein in bladder of rats in acupoint injection group was lower than that in model group(P<0.05),with statistical significance.ConclusionMethylcobalamin Ciliao acupoint injection has a significant therapeutic effect on neurogenic bladder in rats with suprasacral spinal cord injury,which may improve bladder dysfunction and reduce residual urine volume through the SCF/C-kit and JAK2/STAT3 signaling pathways.This study provides a theoretical basis for the clinical application of acupoint injection in the treatment of neurogenic bladder after spinal cord injury.