针刺研究2024，Vol.49Issue(2)：110-118,9.DOI:10.13702/j.1000-0607.20230701
夹脊电针对急性脊髓损伤大鼠脊髓组织NOD样受体热蛋白结构域相关蛋白3介导的细胞焦亡的影响
Effect of electroacupuncture of"Jiaji"(EX-B2)on NOD-like receptor protein 3 mediated pyroptosis in spinal cord tissue of rats with acute spinal cord injury
摘要
Abstract
Objective To observe the effect of electroacupuncture(EA)stimulation of"Jiaji"(EX-B2)on motor function,histomorphology,and expression of NOD-like receptor protein 3(NLRP3)and N-terminal domain of gasdermin D(GSDMD-N)in the spinal cord tissue of rats with spinal cord injury(SCI),so as to explore its mecha-nism underlying improvement of SCI.Methods Forty eight female SD rats were randomly divided into sham surgery(sham),SCI model(model),EA,and NLRP3 agonist(monosodium urate,MSU)combined with Jiaji EA(MSU+EA)groups,with 12 rats in each group which were further divided into 3 d and 7 d subgroups,with 6 rats at each time point.Two EA groups received EA stimulation of EX-B2 with a frequency of 100 Hz,electrical current of 1-2 mA for 30 min,once a day for 3 or 7 days.After 5 min,6 h,and 24 h of modeling,rats of the MSU+EA group received intraperitoneal injection of MSU(200 μg/kg,200 μg/mL).The motor function was evaluated using Basso-Beattie-Bresnahan(BBB)scale,the morphological structure of rat spinal cord tissue was observed by H.E.staining.The expression of pyroptosis related factors NLRP3,cleaved Caspase-1 and GSDMD-N of the spinal cord was observed by using immunohisto-chemistry and Western blot separately,the expression and localization of Iba-1 and GSDMD-N in the spinal cord tissue were observed using immunofluorescence double staining method.Results Compared with the sham group,the BBB scores after modeling and on day 3 and 7 were decreased(P<0.05),while the average OD values(immunoactivity)and expression levels of NLRP3,cleaved Caspase-1 and GSDMD-N proteins,and the immunofluorescence intensity of Iba-1/GSDMD-N(co-expression)of the spinal cord tissues on day 3 and 7 were significantly increased in the model group(P<0.05).In comparison with the model group,the BBB scores on day 3 and 7 were obviously increased(P<0.05),while the immunoactivity and expression levels of NLRP3,cleaved Caspase-1 and GSDMD proteins,and the immunofluorescence intensity of Iba-1/GSDMD-N on day 3 and 7 significantly down-regulated in the EA group(P<0.05)but not in the MSU+EA group(P>0.05),suggesting an elimination of the effects of EA after administration of NLRP3 agonist(MSU).H.E.staining showed obvious bleeding area in the spinal cord tissue,loose tissue and inflammatory cell infiltration on day 3 after modeling,and basic absorption of the bleeding,loose tissue,obvious vacuolar changes of the white matter area,loss and contraction of neurons with infiltration of a large number of inflammatory cells,which was milder in the EA group but not in the MSU+EA group.Conclusion EA of EX-B2 can improve the motor function of SCI rats,which may be related to its functions in inhibiting pyroptosis of microglia mediated by NLRP3/Caspase-1 signaling pathway.关键词
电针/夹脊/脊髓损伤/NOD样受体热蛋白结构域相关蛋白3/细胞焦亡/小胶质细胞Key words
Electroacupuncture/Jiaji(EX-B2)/Spinal cord injury/NLRP3/Pyroptosis/Microglia引用本文复制引用
梅继林,杨志新,李晓宁..夹脊电针对急性脊髓损伤大鼠脊髓组织NOD样受体热蛋白结构域相关蛋白3介导的细胞焦亡的影响[J].针刺研究,2024,49(2):110-118,9.基金项目
黑龙江省头雁团队项目基金(No.2019-9) （No.2019-9）
黑龙江中医药大学校级科技创新研究平台项目(No.2018pt03) （No.2018pt03）
