摘要
目的:探讨小续命汤联合超早期针刺督脉对于脑缺血再灌注模型大鼠自噬相关蛋白核转录因子-κB p65(NF-κB p65)的影响,研究自噬相关蛋白NF-κB p65与脑保护机制的关系,寻找针刺最佳干预时间点。方法:将152只成年SD雄性大鼠随机分为假手术组、模型组、小续命汤高剂量组(药高组)、小续命汤低剂量组(药低组)、针刺组、小续命汤高剂量+针刺组(针高组)和小续命汤低剂量+针刺(针低组)7组。模型组、药高组、药低组、针刺组根据缺血再灌注30 min,2,4,6 h各分为4个亚组,每组6只。造模成功后,根据Zea Longa的神经功能评分,将符合标准大鼠收入相应组别。假手术组仅做颈动脉剥离;模型组仅造模不做任何治疗;小续命汤高低剂量组按照动物体表面积计算,分别给予60 g·kg-1·d-1和15 g·kg-1·d-1灌胃治疗,针刺治疗采用通督调神针刺。连续治疗14 d后进行神经功能评分;运用蛋白免疫印迹法(Western blot)检测大鼠脑组织中自噬相关蛋白NF-κB p65的表达。结果:与假手术组比较,模型组、药高组、药低组、针刺组、针高组和针低组各时间点神经功能缺损评分显著升高(P<0.01);与模型组比较,药高组、药低组、针刺组、针高组和针低组各时间点神经功能缺损评分显著降低(P<0.01);与假手术组比较,模型组、药高组、药低组、针刺组、针高组和针低组各时间点脑组织中NF-κB p65显著升高(P<0.01);与模型组比较,药高组、药低组、针刺组、针高组和针低组各时间点脑组织中NF-κB p65蛋白表达降低(P<0.05),其中以针高组最为显著(P<0.01)。结论:小续命汤联合超早期针刺督脉可以显著改善脑缺血再灌注模型大鼠神经功能缺损情况;小续命汤联合超早期针刺督脉可以抑制脑缺血再灌注模型大鼠自噬相关蛋白NF-κB p65活性,保护脑功能;小续命汤联合超早期针刺督脉在6 h内各组脑保护效应没有显著差异。
Objective:To investigate the effect of Xiao Xumingtang combined with super-acupuncture along governor meridian on autophagy-related protein nuclear factor-kappa B p65(NF-κB p65)in cerebral ischemia-reperfusion model rats,so as to study the relationship between autophagy-related protein NF-κB p65 and brain protection mechanism,and look for the best intervention time point of acupuncture.Method:A total of 152 adult SD male rats were randomly divided into sham operation group,model group,high-dose Xiao Xumingtang group(high-dose drug group),low-dose Xiao Xumingtang group(low-dose drug group)and acupuncture group.There were seven groups including high-dose Xiao Xumingtang+acupuncture group(high acupuncture group)and low-dose Xiao Xumingtang+acupuncture(low acupuncture group).Model group,highdose drug group,low-dose drug group,and acupuncture group were divided into 4 subgroups according to 30 minutes,2,4,6 h of ischemia-reperfusion,with 6 animals in each group.After successful modeling,according to Zea Longa’s neural function score,eligible rats were included into the corresponding groups.The sham operation group only received carotid artery dissection,the model group was only modeled without any treatment,high and low-dose Xiao Xumingtang groups were calculated based on the body surface area of the animal and given 60 g·kg-1·d-1 and 15 g·kg-1·d-1 drug by gavage for treatment,acupuncture was performed to smooth governor meridian and regulate the mind.After 14 days of consecutive treatment,neurological function was scored.Western blot was used to detect the expression of autophagy-related protein NF-κB p65 in rat brain tissue.Result:Compared with the sham operation group,the neurological impairment scores of the model group,the high-dose drug group,the low-dose drug group,the acupuncture group,the high acupuncture group,and the low acupuncture group were significantly increased at each time point(P<0.01).The neurological impairment scores were significantly lower at each time point than those of the high-dose drug group,low-dose drug group,acupuncture group,high acupuncture group,and low acupuncture group(P<0.01),compared with the sham operation group,NF-κB p65 in model group,high-dose drug group,low-dose group,acupuncture group,high acupuncture group and low acupuncture group was significantly increased in the brain tissue at each time point(P<0.01),compared with the model group,the expression of NF-κB p65 protein in the brain tissue of model group,high-dose drug group,low-dose group,acupuncture group,high acupuncture group and low acupuncture group was decreased at each time point(P<0.05),particular in the high acupuncture group(P<0.01).Conclusion:Xiao Xumingtang combined with ultra-early acupuncture along governor meridian can significantly alleviate neurological impairment in rats with cerebral ischemia-reperfusion model.Xiao Xumingtang combined with ultra-early acupuncture along governor meridian can inhibit cerebral ischemiareperfusion model rats.The activity of autophagy-related protein NF-κB P65 protects the brain function.There is no significant difference in the brain protective effect of Xiao Xumingtang combined with ultra-early acupuncture along governor meridian within 6 hours.
作者
吴海洋
王颖
WU Hai-yang;WANG Ying(The Second Affiliated H ospital of A nhui University ofTraditional Chinese Medicine,Hefei 230000,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2020年第18期30-35,共6页
Chinese Journal of Experimental Traditional Medical Formulae
基金
安徽高校自然科学研究项目(KJ2019A0475)。