摘要
目的:观察针康法对缺氧缺血性脑损伤(HIBD)新生大鼠神经元NLRP3炎症小体及细胞焦亡的影响,探讨其改善缺氧缺血性脑损伤后运动功能缺损的可能机制。方法:选择120只7 d龄大鼠分为正常对照组(A组)、模型对照组(B组)、头穴丛刺组(C组)、康复训练组(D组)和针康组(E组),再均分为24 d、7 d和14 d 3个时间点。结扎颈总动脉构建HIBD动物模型。C组针刺百会以及左右各旁开2 mm处,1次/d,每次留针2 h;D组进行转笼转棒训练,1次/d,10 min/次;E组在头穴丛刺的基础上进行转笼转棒训练。在各时间点,采用网屏实验评定运动功能缺损,TUNEL+Caspase-1免疫荧光双标法观察海马区神经细胞焦亡的情况,蛋白印迹法检测海马区NLRP3、Caspase-1和GSDMD-N端蛋白表达。结果:相较于A组,经缺氧缺血造模处理的4组在各时间点网屏实验评分和海马区神经细胞焦亡率均显著升高,NLRP3、Caspase-1和GSDMD-N端蛋白的表达显著增加,差异具有统计学意义(P<0.05);相比较B组,各治疗组在各时间点网屏实验评分和海马区神经细胞焦亡率均显著减少,NLRP3、Caspase-1和GSDMD-N端蛋白的表达水平降低,其中E组的疗效最明显,差异具有统计学意义(P<0.05)。结论:针康法可改善HIBD新生大鼠的运动功能缺损,且疗效优于单纯针刺或康复疗法,其机制可能与抑制NLRP3炎症小体介导的细胞焦亡有关。
Objective:To observe the effects of the acupuncture-rehabilitation therapy on NLRP3 inflammasome and pyroptosis of neurons in neonatal rats with hypoxic-ischemic brain injury,and to explore its possible mechanism in improving motor function deficit after hypoxic-ischemic brain injury.Methods:120 rats(7 d)were divided into the normal control group(group A),the model control group(group B),the scalp clustering needling group(group C),the rehabilitation training group(group D)and the acupuncture-rehabilitation group(group E),and all groups were subdivided into three time points of 24 d,7 d and 14 d.The model of HIBD was constructed by ligating the common carotid artery.Group C was treated with needling Baihui(DU20)and 2 mm apart on the left and right sides,once a day,and the needle was left for 2 h each time;group D underwent cage rotation and stick rotation training,10 min/time,once a day;group E was treated with scalp clustering needling on the treatment basis of group D.At each time point,the motor function deficit was assessed by the screen test,the focal neuronal pyroptosis in the hippocampal region was observed by the TUNEL+Caspase-1 immunofluorescence double-labeling method,and the expressions of NLRP3,Caspase-1 and GSDMD-N-terminal protein in the hippocampal region were detected by Western blot method.Results:Compared with those in group A,the screen test scores,the neuronal pyroptosis rate in hippocampal region,and the expressions of NLRP3,Caspase-1 and GSDMD-N-terminal protein were all increased significantly at each time point in the other four groups(P<0.05);compared with those in group B,the screen test scores,the neuronal pyroptosis rate in hippocampal region,and the expressions of NLRP3,Caspase-1 and GSDMD-N-terminal protein were decreased in group C,group D and group E at each time point,and the decreases were more significant in group E(P<0.05).Conclusion:The acupuncture-rehabilitation therapy can improve the motor function deficits in HIBD neonatal rats,the efficacy is better than that of acupuncture or rehabilitation therapy alone,and the mechanism may be related to the inhibition of NLRP3 inflammasome-mediated pyroptosis.
作者
刘波
丁小连
单昊宇
康玉莹
白露
LIU Bo;DING Xiaolian;SHAN Haoyu;KANG Yuying;BAI Lu(The Second Affiliated Hospital of Heilongjiang University of Chinese Medicine,Harbin 150001,China;Heilongjiang University of Chinese Medicine,Harbin 150040,China;Qiqihar Medical University,Qiqihar 161003,China)
出处
《针灸临床杂志》
2024年第1期83-89,共7页
Journal of Clinical Acupuncture and Moxibustion
基金
黑龙江中医药大学附属第二医院孙申田青年人才基金,编号:2020KY-14
黑龙江中医药大学研究生创新基金,编号:2022yjscx052。
