Research On The Mechanism Of Jia Ji Electroacupuncture To Inhibit Axonal Growth Of Rho-ROCKⅡ Signaling Pathway Based On ASCI Rats Micro Environment

Object:To investigate the therapeutical mechanism of Jiaji points electro-acupuncture after acute spinal cord injury,we used rat acute spinal cord injury(ASCI)model to observe the expression and variation of NogoA,RhoA,RockⅡ and MLC,which are the axonal regeneration of nerve cells and related inhibitory factors,through the Rho/RockⅡ signaling pathway perspective.Method:The rats were randomly divided into five groups:control group(only used for western blot),model control(ASCI)group,the sham operation control group,Jiaji points electroacupuncture(EA)group and inhibitor group.ASCI rats model was established by the Allen’s method.The BBB scores were record to measure the motor function after the acute spinal cord injury and the pathologic features of spinal cord injury were assessed by the hematoxylin-eosin stain.We use the nerve tracer BDA to assess the variation of the neuron on the neural pathways after acute spinal cord injury.The method of immunohistochemistry and western blot are used to measure the expression of NogoA,RhoA,RockⅡ and MLC.Results:1.It was observed that in the 14 days and 28days after operation,the motor function of the rat in EA group was better than the model control group.And there is a significantly improvement of the BBB scores in the 28 days compared with the 14 days(P<0.05).2.Compared with the model control group,the EA group and inhibitor group had better structure and morphous of neural cellular of rats,especially in the 28 days after operation.3.In the brain and T8-T9 plane of spinal,there were massive staining cells with brownish yellow particles in each group,but there was no significant difference between them(P>0.05),In the T12-L1 plane of spinal,the IOD between the ASCI group,the sham operation control group,EA group and inhibitor group were significant.There is still large of BDA tagged neural cells in the sham operation group,but there was no significant difference(P>0.05).In the ASCI group,compared with the sham operation group,there was significant decrease of the BDA tagged neural cells(P<0.05).Compared with the inhibitor group,there was a significant increase of the regeneration of neurons tagged by the BDA in EA group(P<0.05).4.Immunohistochemical results showed that compare to the ASCI group,there was a significant decrease of the expression of NogoA,RhoA,RockⅡand MLC in the EA group and inhibitor group,respectively(P<0.05).And compare to the 14 days,there was a significant decrease of the expression of NogoA,RhoA,RockⅡ and MLC in the 28 days after operation(P<0.05).5.Compare to the model control group measured by western blot,there was a significant decrease of the expression of NogoA,RhoA,RockⅡ and MLC in the EA group and inhibitor group respectively(P<0.05).Compare to the 14 days,there was a significant decrease of the expression of NogoA,RhoA,ROCKⅡ and MLC in the 28 days after operation(P<0.05).Conclusion:1.The rats cute spinal cord injury could induce the release of the myelin associated inhibitory factor and inhibit the axonal regeneration in the microenvironment of the injury area,finally contribute to the secondary spinal cord injury.2.In the injury area of ASCI,myelin associated inhibitory factor were activated and promoted the expression of RockⅡ,then inhibit the axonal regeneration through the Rho/RockⅡ signal pathway.3.The RockⅡ inhibitor Fasudil could reduce the disorder of the axonal regeneration and the pathological lesion,and promote the functional rehabilitation of neurons by inhibiting the activity of the RockⅡ and its substrate.4.Electroacupuncture can inhibit the expression of the inhibitor like RhoA,NogoA,RockⅡ and MLC in the microenvironment of acute spinal cord injury,and promote the regeneration of axons,relieve the secondary pathological damage of spinal cord,fininally promote the recovery of damaged nerve function.