益元灸通过AMPK-Cx43途径改善脊髓损伤后神经源性膀胱大鼠储尿功能的机制研究OA北大核心CSTPCD

Objective To investigate the effect and mechanism of Yiyuan moxibustion on urinary storage function in rats with neurogenic bladder(NB)after suprasacral spinal cord injury(SCI)based on adenosine activated protein kinase(AMPK)-connexin 43(Cx43)pathway.Methods Twelve female Sprague-Dawley rats were selected as the sham operation group using the random number table method,and 68 rats were used to prepare the suprasacral SCI model according to the modified Hassan Shaker spinal cord transection method.After the SCI model was stable,the standard NB model after SCI was screened out.The rats with successful modeling were divided into the model group(n=14),the Yiyuan moxibustion group(n=14),the inhibitor group(n=14),and the Yiyuan moxibustion+inhibitor group(n=14)according to the random number table method.The rats in the Yiyuan moxibustion group were treated with Yiyuan moxibustion.The rats in the inhibitor group were injected with AMPK inhibitor dorsomorphin(0.2 mg/kg)by tail vein.The rats in the Yiyuan moxibustion+inhibitor group were treated with Yiyuan moxibustion combined with tail vein injection of dorsomorphin(0.2 mg/kg).The sham operation group and the model group did not receive any intervention for 14 days.After the intervention,the bladder function of rats was evaluated by urodynamics.HE staining was used to observe morphological changes of the bladder tissue;the content of adenosine triphosphate(ATP)in bladder tissue was detected by colorimetry.The positive expressions of phosphorylated AMPK(p-AMPK),Cx43 and tyrosine protein kinase receptor(C-kit)in bladder tissue were detected by immunohistochemistry.The protein expressions of AMPK,p-AMPK,Cx43 and C-kit in bladder tissue were detected by Western blotting.Results Compared with the model group,the maximum bladder capacity and bladder compliance of the rats in the Yiyuan moxibustion group were increased,the leak point pressure was decreased,the cells in the Yiyuan moxibustion group were relatively regular and orderly,the vacuolar cells were reduced,and the tissue edema was reduced;the ATP content in the bladder tissue was decreased,the positive expression rate of p-AMPK was increased,the positive expression rate of Cx43 and C-kit were decreased,the protein expressions of p-AMPK protein was increased,the expression of Cx43 and C-kit were decreased,and the mRNA expression of Cx43 and C-kit were decreased(P<0.05).Compared with the Yiyuan moxibustion+inhibitor group,the maximum bladder capacity and bladder compliance of the rats in the Yiyuan moxibustion group increased,the leak point pressure decreased,the cells in the Yiyuan moxibustion group were arranged more closely and the tissue edema was reduced;the ATP content in the bladder tissue decreased,the positive expression rate of p-AMPK increased,the positive expression rates of Cx43 and C-kit decreased,the protein expression of p-AMPK increased,the protein expressions of Cx43 and C-kit decreased,and the mRNA expressions of Cx43 and C-kit decreased(P<0.05).Conclusion Yiyuan moxibustion can improve the urinary storage function of NB rats after suprasacral SCI,and its mechanism may be related to activating the AMPK-Cx43 pathway in bladder tissue,reducing the excitation transmission between bladder detrusor cells,thereby reducing the frequency of smooth muscle contraction.