针灸与康复疗法干预脑缺血模型大鼠神经功能及肠道菌群的变化

Acupuncture and moxibustion combined with rehabilitation therapy improve neurological function and intestinal flora following cerebral ischemia in rats

背景:针灸与康复训练均能够有效缓解脑缺血患者症状,目前有关两者联合治疗是否对脑缺血患者神经功能障、肠道菌群失调具有调控作用尚不明确。目的:探究针灸联合康复疗法对脑缺血大鼠神经功能、肠道菌群的影响。方法:将60只SD大鼠按随机数表法随机分为假手术组、模型组、针刺组、康复组和针康组(n=12)。除假手术组外,其余各组建立脑缺血大鼠模型;假手术组仅分离左侧颈总动脉;针刺组用头穴丛刺干预,康复组进行任务导向性跑台训练,针康组同时给予头穴丛针刺和任务导向性跑台训练干预,干预14 d。观察造模后4 h及1,7,14 d各组大鼠神经功能情况,检测干预14 d后大鼠脑含水量变化,Western blot检测缺血半暗带神经元生长相关蛋白(生长相关蛋白43、神经丝蛋白200、排斥指导分子a)表达,试剂盒检测脑组织乳酸脱氢酶、超氧化物歧化酶、丙二醛水平,酶联免疫吸附法检测脑组织肿瘤坏死因子α、白细胞介素1β质量浓度,实时荧光定量PCR检测大鼠粪便中大肠杆菌、双歧杆菌、乳酸杆菌和肠球菌变化。结果与结论:①造模后4 h及1,7,14 d,假手术组无神经功能障碍,模型组大鼠出现明显神经功能障碍(P<0.05);②与同时段模型组相比,针刺组和康复组大鼠神经功能缺损随着干预时间延长有所改善(P<0.05),而针康组大鼠神经功能改善更显著(P<0.01);③与模型组比较,针刺组、康复组大鼠缺血半暗带生长相关蛋白43、神经丝蛋白200表达上调(P<0.05),排斥指导分子a表达下调(P<0.05);大鼠脑组织含水量和脑组织中乳酸脱氢酶、丙二醛、肿瘤坏死因子α、白细胞介素1β质量浓度降低(P<0.05),超氧化物歧化酶活性升高(P<0.05);粪便中大肠杆菌、肠球菌数量明显增多(P<0.05),双歧杆菌、乳酸菌数量明显降低(P<0.05);且针康组上述指标改善更为显著(P<0.01);④提示针灸联合康复疗法可通过刺激缺血半暗带区神经元再生改善脑缺血大鼠神经功能,减轻氧化应激反应和炎症反应,调节肠道菌群紊乱,对缺血性脑卒中具有保护作用。

BACKGROUND:Both acupuncture and moxibustion and rehabilitation training can effectively alleviate the symptoms of patients with cerebral ischemia.At present,it is unclear whether the combined treatment of the two has a regulatory effect on neurological dysfunction and intestinal flora imbalance in patients with cerebra ischemia and its role mechanism.OBJECTIVE:To explore the effect of acupuncture and moxibustion combined with rehabilitation therapy on neurological function and intestinal flora in rats with cerebral ischemia.METHODS:Sixty Sprague-Dawley rats we re randomly divided into sham ope ration group,model group,acupuncture group,rehabilitation group,and acupuncture+rehabilitation group(n=12 per group).Animal models of cerebral ischemia were prepared in all groups except for the sham operation group.In the sham operation group,the left common carotid artery was only separated but not ligated.The acupuncture group was treated with scalp cluster acupuncture intervention,the rehabilitation group was given task-oriented treadmill training,and the acupuncture+rehabilitation group was given scalp cluster acupuncture and task-oriented treadmill training intervention.Treatments in each group lasted for 14 days.The sham operation and model groups were not inte rvened.The neurological function was assessed by Longa score at 4 hours,1,7,and 14 days after modeling.Changes in brain water content were detected after 14 days of intervention.Western blot was applied to detect the expressions of neuron growth-related proteins(growth-associated protein43,neurofilament 200,repulsive guidance molecule a) in the ischemic penumbra.Kit detection was used for detecting the levels of lactate dehydrogenase,superoxide dismutase,and malondialdehyde in brain tissue.ELISA was applied to detect the levels of tumor necrosis factor-α and inte rleukin-1β in brain tissue.Rea l-time fluorescent qua ntitative PCR was used to detect the changes of Escherichia coli,Bifidobacterium,Lactobacillus,and Enterococcus in rat feces.RESULTS AND CONCLUSION:At 4 hours,1,7,and 14 days after modeling,there was no neurological dysfunction in the sham operation group but obvious neurological dysfunction in the model group(P<0.05).Compared with the model group,the neurological deficits of rats in the acupuncture and rehabilitation gro ups we re significantly improved with the prolonged inte rvention time(P<0.05),and the neurological function of rats in the acupuncture+rehabilitation group improved more significantly(P <0.01).Compared with the model group,the expressions of growth-associated protein 43 and neurofilament 200 in the ischemic penumbra were significantly up-regulated in the acupuncture and rehabilitation groups(P<0.05),the expression of repulsive guidance molecule a was significantly down-regulated(P <0.05),the brain water content and levels of lactate dehydrogenase,malondialdehyde,tumor necrosis factor-α and interleukin-1β in brain tissue were reduced(P<0.01),the superoxide dismutase level was raised(P<0.01),the counts of Escherichia coli and Enterococcus in feces were significantly increased(P<0.01),and the counts of Bifidobocterium and Lactobacillus were significantly decreased(P<0.01),The above-mentioned indicators were improved more significantly in the acupuncture+rehabilitation group(P <0.01).To conclude,acupuncture and moxibustion combined with rehabilitation therapy can improve neurological function,relieve oxidative stress and inflammatory responses,and regulate intestinal flora disorders in rats with cerebral ischemia by stimulating the regeneration of neurons in the ischemic penumbra.The combine therapy has a protective effect against ischemic stroke.