电针联合丰富康复训练对缺血性脑卒中大鼠的神经保护作用：基于NLRC4炎症通路的研究

关键词：缺血性脑卒中  大鼠  NLRC4炎症通路  电针  丰富康复训练  神经细胞  凋亡

Keywords:ischemic stroke  rat  NLRC4 inflammatory pathway  acupuncture  enriched rehabilitation training  nerve cells  apoptosis

摘要：

      摘要 目的：观察电针联合丰富康复训练对缺血性脑卒中（MCAO）大鼠NLRC4/Caspase-1/GSDMD炎症通路的影响，探讨其改善缺血性脑卒中后神经功能的机制。 方法：实验设计采用60只SD雄性大鼠，实验将大鼠分为5组，每组12只：假手术组（Sham组）、大脑中动脉闭塞组（MCAO组）、电针刺激组（EA组）、丰富康复训练组（RT组）及联合治疗组（EA+RT组）。Sham组仅进行颈总动脉分离，其余4组进行大脑中动脉闭塞处理。Sham组和MCAO组不予治疗，EA组分别在“百会”穴和“大椎”穴位给予电针刺激，RT组接受丰富康复训练治疗，EA+RT组则同时接受电针和丰富康复训练治疗。在建模后24h、48h、7d对各组进行Longa、mNSS神经功能评分；实验结束后，使用TUNEL、ELISA、Western Blot和RT-qPCR等方法检测缺血侧脑组织神经细胞凋亡情况、炎症因子IL-18、IL-1β表达水平、炎症通路相关因子NLRC4、GSDMD、Caspase-1的mRNA及蛋白的表达情况。 结果：相比于Sham组，其余4组建模后各时期Longa、mNSS评分和细胞凋亡率均升高，相比于MCAO组，各治疗组评分和神经细胞凋亡率均明显下降（P<0.05），且EA+RT组下降更为显著（P<0.05）。与Sham组相比，MCAO组NLRC4、Caspase-1、GSDMD的mRNA和蛋白表达以及IL-18、IL-1β表达均明显升高（P<0.05）；与MCAO组相比，各治疗组上述指标均明显下降（P<0.05），其中EA+RT组下降更为显著（P<0.05）。 结论：电针联合丰富康复训练可以抑制缺血区脑组织NLRC4/Caspase-1/GSDMD炎症通路的表达从而改善运动功能障碍。

Abstract:

      Abstract Objective：To observe the effect of electroacupuncture (EA) combined with enriched rehabilitation training (RT) on the NLRC4/Caspase-1/GSDMD inflammatory pathway in rats with ischemic stroke （MCAO）， and to explore the underlying mechanisms of neurological recovery. Method：Sixty male SD rats were randomly assigned to five groups (n=12 in each group)： Sham operation group （Sham group）， middle cerebral artery occlusion group （MCAO group）， electroacupuncture stimulation group （EA group）， rich rehabilitation training group （RT group）， and combined treatment group （EA+RT group）. Only common carotid artery separation was performed in the Sham group，while the other four groups were treated with middle cerebral artery occlusion. The Sham group and the MCAO group were not treated. The EA group was given electroacupuncture stimulation at the "Baihui" and "Dazhui" acupoints respectively. The RT group received enriched rehabilitation treatment. The EA+RT group received both electroacupuncture and rich rehabilitation treatment simultaneously. The Longa and mNSS neurological function scores of each group were conducted at 24 hours， 48 hours and 7 days after modeling. After the experiment， TUNEL， ELISA and Western Blot were used. The apoptosis of nerve cells in the ischemic brain tissue，the expression levels of inflammatory factors IL-18 and IL-1β， and the mRNA and protein expression of inflammatory pathway-related factors NLRC4， gasdermin D (GSDMD)， and Caspase-1 were analyzed by RT-qPCR and Western Blot. Result：Compared with the Sham group， MCAO induced significant increases in Longa and mNSS scores and neuronal apoptosis at all time points. Compared with the MCAO group， all treatment groups showed significant reductions in neurological deficit scores and neuronal apoptosis （P<0.05）， with the EA+RT group demonstrating the greatest improvement（P<0.05）. Compared with the Sham group， expression levels of NLRC4, Caspase-1, GSDMD, IL-18, and IL-1β were significantly elevated in the MCAO group（P<0.05）. These markers were significantly reduced in all treatment groups compared with the MCAO group (P<0.05), with the most pronounced reductions observed in the EA+RT group (P<0.05). Conclusion： Electroacupuncture combined with enriched rehabilitation training can inhibit the expression of the NLRC4/Caspase-1/GSDMD inflammatory pathway in ischemic brain tissue， thereby improving motor dysfunction.