交叉电项针对脑出血延髓损伤模型豚鼠延髓组织cAMP-PKA通路的影响  

Effect of Cross Electro-nape-acupuncture on cAMP-PKA Pathway in Medullary Tissue of Cerebral Hemorrhage Bulbar Injury Model Guinea Pigs

作　　者：贾坤平 梁群 刘关平[1,2] 蔡国锋 吴建丽[1] JIA Kunping;LIANG Qun;LIU Guanping;CAI Guofeng;WU Jianli(Heilongjiang University of Chinese Medicine,Harbin,150040;Hanan Branch of the Second Affiliated Hospital of Heilongjiang University of Chinese Medicine;The First Affiliated Hospital of Heilongjiang University of Chinese Medicine)

机构地区：[1]黑龙江中医药大学,黑龙江省哈尔滨市150040 [2]黑龙江中医药大学附属第二医院哈南分院 [3]黑龙江中医药大学附属第一医院

出　　处：《中医杂志》2023年第11期1157-1164,共8页Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金(82105030);教育部“春晖计划”合作科研项目(HLJ2019031);黑龙江中医药大学科研基金(2019MS19)。

摘　　要：目的探讨交叉电项针对脑出血延髓损伤豚鼠神经功能的影响及可能作用机制。方法240只豚鼠随机分为空白组、模型组、针刺组、交叉电项针组和电项针组,每组又分为干预1、3、7、14天4个时间点,每个时间点各12只。除空白组外,其余各组豚鼠采用自体血注入法制备脑出血延髓损伤模型。造模12 h后开始干预,针刺组豚鼠选取双侧“风池”“翳风”针刺,每次20 min,每日1次。交叉电项针组豚鼠以电针仪电极在项部交叉连接双侧“风池”“翳风”,电项针组豚鼠电针电极不交叉连接双侧“风池”“翳风”,均每次20 min,每日1次。空白组、模型组豚鼠给予同样的抓取固定,但不干预。分别于干预1、3、7、14天采用改良神经功能缺损量表(mNSS)评定豚鼠神经功能缺损程度;各时间点取豚鼠延髓组织免疫组织化学法检测突触生长相关蛋白43(GAP-43)、突触素(SYN)阳性表达,ELISA法检测环磷酸腺苷(cAMP)、蛋白激酶A(PKA)含量,Western blot法检测cAMP蛋白及磷酸化环磷酸腺苷反应元件结合蛋白(p-CREB)表达。结果空白组豚鼠无神经损伤。与空白组同时间比较,其余各组豚鼠在各时间点mNSS评分均显著增加(P<0.01);与模型组同时间比较,针刺组、电项针组和交叉电项针组豚鼠在干预3、7、14天mNSS评分均有不同程度降低(P<0.01),各干预组间各时间点比较差异无统计学意义(P>0.05)。与空白组同时间比较,模型组豚鼠延髓组织各时间点GAP-43阳性表达升高,SYN阳性表达降低,cAMP、PKA含量及cAMP、p-CREB蛋白表达均降低(P<0.05或P<0.01)。与模型组同时间比较,各干预组在干预3、7、14天GAP-43、SYN阳性表达,cAMP、PKA含量及cAMP、p-CREB蛋白表达均升高(P<0.05或P<0.01),且交叉电项针组在干预7、14天时各指标改善优于针刺组和电项针组(P<0.05或P<0.01)。结论交叉电项针可改善脑卒中延髓损伤豚鼠神经功能缺损情况,其机制可能与Objective To observe the effect and possible mechanism of cross electro-nape-acupuncture(CENA)on neurological function of guinea pigs with cerebral hemorrhage bulbar injury.Methods A total of 240 guinea pigs were randomly divided into blank group,model group,acupuncture group,CENA group and electro-nape-acupuncture(ENA)group,and each group was further divided into 1-,3-,7-and 14-day intervention timepoints,with 12 guinea pigs at each timepoint.The model of cerebral hemorrhage bulbar injury was established by injection of the guinea pigs'autologous blood in all the groups except for the blank group.Twelve hours after modelling,the guinea pigs in the acupuncture group were needled at bilateral“Fengchi(GB 20)”and“Yifeng(SJ 17)”,20 minutes each time,once a day.In CENA group,the positive pole of the needle was connected to“Fengchi(GB 20)”,while the negative pole was connected to the opposite side of“Yifeng(SJ 17)”;in ENA group,the positive pole of the needle was connected to“Fengchi(GB 20)”,while the negative pole was connected to“Yifeng(SJ 17)”on the same side;the intervention was applied 20 minutes each time,once a day.The guinea pigs in the blank group and the model group were given the same grabbing and fixation without intervention.After 1-,3-,7-and 14-day intervention,the neurologic function was assessed by using modified neurological severity scale(mNSS).Positive expression of synapse growth-associated protein 43(GAP-43)and synaptophysin(SYN)in guinea pig medullary tissue were detected by immunohistochemistry at each timepoint.Contents of cyclic adenosine monophosphate(cAMP)and protein kinase A(PKA)were detected by ELISA.The protein expressions of cAMP and phosphorylated cyclic AMP reponse response element binding protein(p-CREB)were detected by Western blot.Results The guinea pigs in the blank group had no neurological deficit.Compared to those in the blank group,the mNSS score at different timepoints in the other groups significantly increased(P<0.01).The mNSS score showed different de

关 键 词：脑出血 延髓损伤 交叉电项针 突触可塑性 cAMP-PKA通路 

分 类 号：R245[医药卫生—针灸推拿学]