Mechanism Of Jiaji Electroacupuncture Regulating ASC To Improve Inflammatory Response In Rats With Spinal Cord Injury

Objective:To investigate the effects of Jiaji electroacupuncture on motor function and spinal cord histomorphology in rats with acute spinal cord injury(ASCI);To investigate the effects of Jiaji electroacupuncture on the expression of Apoptosis-associated speck-like protein containing a CARD(ASC)in the spinal cord of rats with ASCI;To investigate whether Jiaji electroacupuncture regulates the degree of inflammatory injury after ASCI in rats through the NOD-like receptor protein 3(NLRP3)signaling pathway.In order to provide theoretical basis for clinical practice.Methods:Experiment 1: Effects of Jiaji electroacupuncture on motor function and spinal cord histomorphology in rats with ASCIThirty adult healthy Sprague Dawley(SD)female rats were randomly divided into five groups:(1)sham-operated group(Sham group,n=6)(2)model group(Model group,n=6)(3)Jiaji electroacupuncture group(EA group,n=6)(4)monosodium urate group(MSU group,n=6)(5)monosodium urate + Jiaji electroacupuncture group(MSU + EA group,n=6).Each group was measured at 3d and 7d after the intervention,and the subgroup at each time point was 3 rats.Sham group: Only the spinal cord tissue was exposed and sutured immediately without any other disposition,and the rats were kept routinely after the operation;Model group: After successful modeling,they were kept routinely without any other disposition;EA group: 3 hours after successful modeling,the rats were treated with electroacupuncture intervention after awakening,once a day until completion of the assay at each time point.MSU group: Rats were injected with 200μg/m L/Kg MSU intraperitoneally once at 5min,6h and 24 h after successful modeling;MSU+EA group: Rats were injected with 200μg/m L/Kg MSU intraperitoneally once at 5min,6h and 24 h after successful modeling,and were also treated with electroacupuncture.(MSU is NLRP3 activator)Experiment 2: Immunohistochemical method(IHC)was used to detect the content of ASC protein in spinal cord tissue of rats in each groupThirty adult SD rats were randomly divided into five groups: sham-operated group,model group,Jiaji electroacupuncture group,monosodium urate group and monosodium urate agent + Jiaji electroacupuncture group.The modeling method and intervention method were the same as in Experiment 1.After the rats in each group were taken,the ASC protein content in the spinal cord tissue was detected by IHC.Results:1.BBB score:The BBB scores of rats in Sham group were 21 at 3d and 7d,and no motor dysfunction was observed.3d and 7d postoperatively,the BBB scores of rats in Model,EA,MSU and MSU+EA groups decreased significantly compared with Sham group,among which the hind limb dysfunction of rats in Model and MSU groups was serious,and the difference was statistically significant(P < 0.05).The BBB scores of rats in the EA group and MSU+EA group showed an increasing trend at 3d and 7d postoperatively,and compared with the Model group,the BBB scores of rats in the EA group increased significantly,while the BBB scores of rats in the MSU group decreased significantly,and the BBB scores of rats in the MSU+EA group increased significantly compared with the MSU group,and the differences were statistically significant(P < 0.05).It showed a trend of increasing the score with longer treatment time.2.HE results:After HE staining,under electron microscopic observation,the spinal cord tissue sections of the Sham group at each time point had clear gray and white matter boundaries,intact neuronal cell structure,dense arrangement,uniform cell distribution,no obvious hemorrhage,no neuronal cell swelling and tissue degeneration,no neuroedema,and clearly visible cell nuclei located in the center of the cells and rounded morphology.At 3d postoperatively,the Model group showed tissue congestion,blurred gray and white matter boundaries,inflammatory infiltration,neuronal cell loss and crinkling,and formation of vacuoles.Compared with the Model group,the EA group had more intact nerve cell structure,less inflammatory cells,a small amount of congestion,and less neuronal crinkling,while the MSU group had severe hemorrhage,a large amount of inflammatory cell infiltration,and more severe neuronal crinkling loss.the MSU+EA group had congested spinal cord tissue,inflammatory cell infiltration,and neuronal crinkling,and the degree of injury tended to be similar to that of the Model group.At 7d after surgery,the rats in the Model and MSU groups had sparse edema and still had a large amount of inflammatory infiltration.Compared with them,the cell structure and morphology of the EA group partially tended to be normal,the area of inflammatory infiltration was reduced,and the neuronal cells were obviously increased,but their morphological structure was irregular;the MSU+EA group had more complete cell structure and morphology than the MSU group,but the area of inflammatory infiltration was larger than that of the EA group.3.IHC:The IHC results showed that compared with the Sham group,the expression of ASC in the spinal cord tissues of rats in the Model,EA,MSU and MSU+EA groups was increased at3 d and 7d.The MSU group showed the most significant increase,followed by the Model group,and the difference was statistically significant(P < 0.05);at 3d and 7d after surgery,the expression of ASC in the EA group decreased compared with the Model group,and the expression of ASC in the MSU+EA group decreased compared with the MSU group.The differences were statistically significant(P < 0.05).Conclusion:1.Jiaji electroacupuncture can improve the motor function of the limbs and promote the recovery of motor function of the lower limbs in ASCI rats.2.Jiaji electroacupuncture may reduce ASC expression in spinal cord tissues by regulating the NLRP3 signaling pathway,thereby reducing histopathological inflammatory changes in the spinal cord of ASCI rats.