| Objective: To investigate the effects of electroacupuncture(EA)on neurogenic bladder(NB)after complete suprasacral spinal cord injury(SSCI)in rats and its mechanism by observing the changes of urodynamics(including maximum bladder capacity,bladder base pressure,leak point pressure and bladder compliancy),morphological changes of the detrusor muscle,the expression of PACAP and its mRNA,cAMP,PKA,p-MLCK,p-MLCP and p-MLC in detrusor muscle,meanwhile blocking PACAP-cAMP-PKA signaling pathway.Methods:This study is composed of three parts.In the first part,SD rats were randomly divided into 4 groups,normal control group,sham operated control group,model group and EA treatment group.The spinal cord injury model on T10/T11 was made with modified spinal cord transaction.Urodynamics before and after treatment were examined,such as maximum bladder capacity,bladder base pressure,leak point pressure and bladder compliancy.At the end of experiment,the morphological changes of the detrusor muscle were examined by light microscopy.In the second part,the SD rats were randomly divided into 5 groups,normal control group,sham operated control group,model group,EA treatment group and EA +PACAP6-38 treatment group.The methods of model making,EA and urodynamic tests were the same as the first part.The expression of PACAP38,PACAP38 m RNA,and cAMP in detrusor muscle were determined by immunohistochemistry,semi-quantitative reverse transcription polymerase chain reaction(RT-PCR)and western blot.In the third part,the SD rats were randomly divided into 5 groups,normal control group,sham operated control group,model group,EA treatment group and EA + H89 treatment group.The methods of model making,EA and urodynamic tests were the same as the first part.The expression of PKA,p-MLCK,p-MLCP and p-MLC in detrusor muscle was determined by western blot and ELISA.Results:(1)Compared with the control group,maximum bladder capacity significantly decreased(P<0.01),bladder base pressure and leak point pressure obviously increased(P<0.01),and bladder compliancy decreased(P<0.01)in the model group.After EA treatment,maximum bladder capacity and bladder compliancy significantly decreased(P<0.01),bladder base pressure and leak point pressure obviously increased(P<0.01)in EA group(P<0.05 or P<0.01).(2)Compared with the EA group,maximum bladder capacity and bladder compliancy significantly increased(P<0.05)both in EA+ PACAP6-38 group and EA+ H89 group after intraperitoneal injection of PACAP6-38 or H89 in SD rats.(3)Compared with the model group,Zhongji-Ciliao-Sanyinjiao-Dazhui EA can significantly up-regulate the expression of PACAP38,incresease the content of cAMP in the detrusor of rats(P<0.05 or P<0.01).Compared with the control group and the EA group,the content of cAMP decreased slightly(P > 0.05)after intraperitoneal injection of PACAP6-38 in EA+ PACAP6-38 group rats.(4)Compared with the model group,Zhongji-Ciliao-Sanyinjiao-Dazhui EA can significantly up-regulate the expression of PKA,p-MLCK,p-MLCP,decresease the expression of p-MLC in the detrusor of rats(P<0.05 or P<0.01).Compared with the EA group,the expression of PKA and p-MLCK obviously(P>0.05)decreased after intraperitoneal injection of H89 in EA+ H89 group rats,nearly the levels of the control group.Conclusion:(1)EA can significantly improve the maximum bladder capacity and bladder compliancy,downgrade the bladder base pressure,leak point pressure of rats with NBafter SSCI.(2)Electroacupuncture can promote detrusor relaxation and improve bladder function by increasing the expression of PACAP and up-regulating the content of cAMP in detrusor of rats with NB after SSCI.(3)Electroacupuncture can be beneficial for bladder fuction reconstruction after SSCI via increasing the expression of PKA,promoting the phosphorylation of MLCK and MLCP,increasing the phosphorylation of MLC.(4)EA may be an effective treatment for NB after SSCI via activation of PACAP-cAMP-PKA signaling pathway. |
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