夹脊电针刺激通过PKA/RhoA通路治疗大鼠急性脊髓损伤  

Treatment of acute spinal cord injury in rats with Jiaji electro-acupuncture stimulation via PKA/RhoA pathway

作　　者：周孝敏 鱼丽萍 李卓伦 丁天红[3] ZHOU Xiaomin;YU Liping;LI Zhuolun;DING Tianhong(The Second Clinical Medical Collgeg,Shaanxi University of Traditional Chinese Medicine,Xianyang 712000,China;Department of Neurology,the First People's Hospital Xianyang City,Xianyang 712000,China;The Second Affiliated Hospital of Shaanxi University of Chinese Medicine,Xianyang 712000,China)

机构地区：[1]陕西中医药大学第二临床医学院,咸阳712000 [2]咸阳市第一人民医院神经内科,咸阳712000 [3]陕西中医药大学第二附属医院,咸阳712000

出　　处：《神经解剖学杂志》2023年第3期295-302,共8页Chinese Journal of Neuroanatomy

基　　金：陕西省科技厅重点研发计划项目(2017SF-304);陕西省咸阳市科技局科研项目(2017KO2-78)。

摘　　要：目的:探究电针(EA)夹脊穴对脊髓损伤(SCI)大鼠后肢运动功能及蛋白激酶A(PKA)/Ras同源基因家族成员A(RhoA)通路表达的影响,并从PKA/RhoA通路初步探讨其机制。方法:将36只雌性SD大鼠随机分为假手术组(Sham)、SCI模型组(SCI)、电针干预组(SCI+EA)。运用钳夹法建立SCI模型,SCI+EA组电针干预T_(9)和T_(11)两对夹脊穴。利用脊髓功能评分(BBB量表)检测大鼠的后肢运动功能,HE染色观察脊髓组织形态学改变,同时运用免疫荧光染色、Western Blot及real time RT-PCR检测脊髓组织中蛋白激酶A(PKA)、Ras同源基因家族成员A(RhoA)及生长相关蛋白-43(GAP-43)的表达。结果:SCI模型大鼠后肢运动功能发生严重障碍,脊髓组织结构紊乱,伴较多出血点,而电针刺激后SCI大鼠脊髓组织结构较完整,出血点减少,后肢运动功能也得到明显改善。此外,夹脊电针刺激可显著提高SCI大鼠脊髓组织中PKA mRNA表达,(P<0.05)。同时,夹脊电针刺激后SCI大鼠脊髓组织中PKA、GAP-43蛋白以及磷酸化蛋白p-RhoA(ser188)表达均显著升高(P<0.05)。结论:夹脊电针能改善SCI大鼠后肢运动功能,减轻脊髓组织的病理学损伤;可能是通过调节PKA/RhoA信号通路实现的。Objective:To investigate the effect of electro-acupuncture(EA) at Jiaji points on motor function of hind limbs and expression of protein kinase A(PKA)/Ras homolog gene family member A(RhoA) pathway in rats with spinal cord injury(SCI),and to explore the molecular mechanism of EA treatment on spinal cord injury from PKA/RhoA pathway.Methods:Thirty-six female SD rats were randomly divided into sham group,SCI group,SCI+EA group.The SCI model was established by clamp method,and the rats in the SCI+EA group were treated with EA at T_(9) and T_(11) Jiaji points.Behavioral recovery of rats was scored according to the BBB(Basso,Beattie,Bresnahan) scale,and HE staining was performed to observe pathological changes of injured spinal cord.At the same time,immunofluorescence staining,Western Blot and real time RT-PCR was used to detect the expression of PKA,RhoA and GAP-43.Results:SCI model rats had severe hind limbs motor dysfunction,and the spinal cord tissue structure was disordered with many hemorrhagic spots.However,after treatment with EA,the spinal cord tissue structure was more complete with fewer hemorrhagic spots,and the motor function of the hind limbs was significantly improved.In addition,EA treatment can promote the expression of PKA mRNA in spinal cord tissue of SCI rats(P<0.05).At the same time,the expressions of PKA,GAP-43 protein and phosphorylated protein p-RhoA(ser188) in spinal cord tissue of SCI rats were significantly increased after EA treatment(P<0.05).Conclusion:EA at Jiaji points can improve the motor function of hind limbs and pathological injury of spinal cord tissue in SCI rats,which may be achieved by regulating PKA/RhoA signaling pathway.

关 键 词：脊髓损伤 夹脊穴 电针 蛋白激酶A Ras同源基因家庭成员A 大鼠 

分 类 号：R245[医药卫生—针灸推拿学]