通利枢机针刺法通过Notch通路对脑卒中后肢体痉挛大鼠肢体功能及神经功能的影响

Effect of Unchoking Pivot Acupuncture on Limb and Nerve Function Recovery in Rats with Limb Spasticity After Stroke Through Notch Pathway

目的:探讨通利枢机针刺法通过Notch通路对脑卒中后肢体痉挛大鼠肢体功能及神经功能恢复的影响。方法:从80只SPF级雄性SD大鼠中随机选取24只大鼠作为对照组(n=12)和假手术组(n=12),其余大鼠采用Zea Longa线栓法与内囊注射NMDAR法建立脑卒中后肢体痉挛大鼠模型。造模成功的46只大鼠随机分为模型组(n=11)、DAPT组(n=11)、针刺组(n=12)、针刺+DAPT组(n=12),造模成功后开始给予对应干预,针刺组大鼠取左侧天井、正营、环跳穴位进行针刺,留针15 min,每5 min施展1次平补平泻手法,每次10 s,每日1次;DAPT组大鼠以0.1 mg·kg^(-1)剂量腹腔注射DAPT溶液,每日1次;针刺+DAPT组干预手段同上述针刺组、DAPT组;假手术组、模型组大鼠在相同时间仰卧位固定于鼠板上,不进行针刺治疗,腹腔注射生理盐水;对照组不捆绑、不治疗,持续2周。Zea Longa评分评估各组大鼠神经功能;采用Feeney平衡木评分评价大鼠肢体功能;使用电生理描记结果评估肌张力;Western Blot测定脑组织Notch1、Jagged1、γ-氨基丁酸a型(γ-aminobutyricacid type A,GABAa)、γ-氨基丁酸转运体(γ-aminobutyric acid transporter-1,GAT-1)的蛋白表达水平;RT-PCR检测Notch1 mRNA、Jagged1 mRNA、GABAa mRNA、GAT-1 mRNA表达水平。结果:通利枢机针刺法能显著降低脑卒中后肢体痉挛大鼠Zea Longa评分、Feeney平衡木评分(P<0.05),显著提高电生理描记结果(P<0.05),显著升高Notch1、Jagged1、GABAa蛋白及mRNA表达水平(P<0.05),显著降低GAT-1蛋白及mRNA表达水平(P<0.05)。结论:通利枢机针刺法可提高脑卒中后肢体痉挛大鼠神经功能,改善肢体功能,其作用可能与调控Notch通路有关。

Objective: To explore the effect of unchoking pivot acupuncture on limb spasticity in rats after stroke through Notch pathway Influence of body function and nerve function recovery.Methods: 24 rats were randomly selected from 80 SPF grade male SD rats as control group(n=12) and sham operation group(n=12).The rest rats were used ZeaLonga suture method and internal capsule injection of NMDAR method to establish limb spasm after stroke.46 rats were randomly divided into model group(n=11)(3,5-difluorophenylacetyl)-L-alanyl-L-2-phenylglycine tert-butyl ester[(3,5-Difluorophenacetyl)-L-alanyl-S-phenylglycine-2-butylEster, DAPT] group(n=11),acupuncture group(n=12),acupuncture+DAPT group(n=12),the corresponding intervention began on the first day after the success of the model.The rats in the acupuncture group took the left Tianjing, Zhengying and Huantiao acupoints, left the needle for 15 minutes, and applied the method of flat-reinforcing and flat-reducing once every 5 minutes, 10 seconds each time, once a day;Rats in the DAPT group were intraperitoneally injected with DAPT solution at a dose of 0.1 mg·kg^(-1),once a day;The intervention methods of acupuncture+DAPT group were the same as those of acupuncture group and DAPT group;The sham operation group and the model group were fixed on the rat board in supine position at the same time, without acupuncture treatment, and injected with saline intraperitoneally;The control group did not bind or treat for 2 weeks.ZeaLonga score was used to evaluate the nerve function of rats in each group;Feeney balance beam score was used to evaluate the limb function of rats;The results of electrophysiology were used to evaluate muscle tension;The brain tissue Notch1,Jagged1 γ-Aminobutyric acid type a(GABAa) and γ-Aminobutyric acid transporter(GAT-1) protein expression level.The expression levels of Notch1 mRNA,Jagged1 mRNA,GABA amRNA and GAT-1 mRNA were detected by RT-PCR.Results: Results: Unchoking pivot acupuncture can significantly reduce the ZeaLonga score and Feeney balance beam score of rats with limb spasm after stroke(P<0.05),significantly improved(P<0.05),the expression levels of Notch1,Jagged1,GABAa protein and mRNA were significantly increased(P<0.05),and the expression levels of GAT-1 protein and mRNA were significantly decreased(P<0.05).Conclusion: Unchoking pivot acupuncture can improve the nerve function and limb function of rats with limb spasm after stroke, which may be related to the regulation of Notch pathway.