基于SIRT1/PGC-1α通路探讨电针联合丰富康复训练对脑缺血大鼠氧化应激的影响
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国家自然科学基金项目(81874500);安徽省教育厅自然科学基金重点项目(KJ2019A0430);安徽省自然科学研究青年基金项目(1808085QH268)


Effect of Electroacupuncture Combined with Enriched Rehabilitation Training on Oxidative Stress in Rats with Cerebral Ischemia Based on the SIRT1/PGC-1α Pathway
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    目的 探讨电针联合丰富康复训练对脑缺血大鼠急性期神经损伤的保护机制。方法 随机选取15只SD大鼠为假手术组,另将模型复制成功的大脑中动脉闭塞(middle cerebral artery occlusion,MCAO)模型大鼠60只随机分为模型组、电针组、丰富康复训练组(康复组)和电针联合丰富康复训练组(联合组),每组15只。假手术组和模型组不进行干预,电针组和联合组于“百会”“大椎”进行电针治疗;康复组和联合组予以丰富环境与康复训练,每次30 min,每日1次,连续治疗3 d。观察大鼠缺血侧皮质区脑血流量、组织形态学、阳性细胞率的动态变化,检测缺血侧皮质中丙二醛(malonic dialdehyde,MDA)、超氧化物歧化酶(superoxide dismutase,SOD)含量;RT-PCR检测沉默信息调节因子1(silent information regulator 1,SIRT1)、过氧化物酶体增殖物激活受体γ共激活因子1α(peroxisome proliferator-activated receptor γ coactivator-1α,PGC-1α)、B淋巴细胞瘤-2(B-cell lymphoma-2,Bcl-2)、半胱氨酸蛋白酶-3(cysteinyl aspartate specific proteinase-3,Caspase-3)及半胱氨酸蛋白酶-9(cysteinyl aspartate specific proteinase-9,Caspase-9)mRNA的表达水平。结果 分组因素对缺血侧皮质中MDA、SOD水平和SIRT1、PGC-1α、Caspase-3、Caspase-9、Bcl-2 mRNA表达水平的主效应均有统计学意义(P<0.05)。与假手术组比较,模型组大鼠脑血流量,SOD水平,SIRT1、PGC-1α、Bcl-2 mRNA表达水平均显著降低(P<0.05),阳性细胞率,MDA水平,Caspase-3、Caspase-9 mRNA表达水平均显著升高(P<0.05);与模型组比较,各治疗组SOD水平,SIRT1、PGC-1α、Bcl-2 mRNA表达水平显著上升(P<0.05),MDA水平和Caspase-3、Caspase-9 mRNA表达水平显著降低(P<0.05);联合组与电针组、康复组比较,上述各指标表达水平均有统计学意义(P<0.05);电针联合丰富康复训练对缺血侧皮质MDA、SOD水平和SIRT1、PGC-1α mRNA表达水平的影响具有交互作用(P<0.05),对Caspase-3、Caspase-9、Bcl-2 mRNA表达水平的影响无交互作用(P>0.05)。结论 电针联合丰富康复训练可通过调控SIRT1/PGC-1α通路,提高抗氧化能力,改善神经细胞损伤,发挥神经保护作用。

    Abstract:

    Objective To investigate the protective mechanism of electroacupuncture combined with enriched rehabilitation training against acute nerve injury in rats with cerebral ischemia. Methods A total of 15 Sprague-Dawley rats were randomly selected as sham-operation group, and 60 rats used to establish a model of middle cerebral artery occlusion (MCAO) were randomly divided into model group, electroacupuncture group, enriched rehabilitation training group (rehabilitation group), and electroacupuncture+enriched rehabilitation training group (combination group), with 15 rats in each group. The rats in the sham-operation group and the model group were not given any intervention; the rats in the electroacupuncture group and the combination group were given electroacupuncture at Baihui and Dazhui; the rats in the rehabilitation group and the combination group were given enriched environmental and rehabilitation training for 30 minutes each time, once a day for 3 consecutive days. The dynamic changes of cerebral blood flow, histomorphology, and apoptosis rate in the ischemic cortex were observed, and then the content of malondialdehyde (MDA) and superoxide dismutase (SOD) was measured. RT-PCR was used to measure the mRNA expression levels of silent information regulator 1 (SIRT1), peroxisome proliferator-activated receptor gamma coactivator-1 alpha (PGC-1α), B-cell lymphoma-2 (Bcl-2), Caspase-3,and Caspase-9. Results Grouping factors had a statistically significant main effect on the levels of MDA and SOD in ischemic cortex and the mRNA expression levels of SIRT1,PGC-1α,Caspase-3,Caspase-9,and Bcl-2 (P<0.05). Compared with the sham-operation group,the model group had significant reductions in cerebral blood flow,the level of SOD, and mRNA expression levels of SIRT1, PGC-1α,and Bcl-2 (P<0.05) and significant increases in apoptosis rate, the level of MDA, and mRNA expression levels of Caspase-3 and Caspase-9 (P<0.05). Compared with the model group, the treatment groups had significant increases in the level of SOD and mRNA expression levels of SIRT1, PGC-1α, and Bcl-2 (P<0.05) and significant reductions in the level of MDA and mRNA expression levels of Caspase-3 and Caspase-9 (P<0.05). There were significant differences in the above indices between the combination group and the electroacupuncture group or rehabilitation group (P<0.05). Electroacupuncture combined with enriched rehabilitation training had synergistic interaction on the levels of MDA, SOD and mRNA expression levels of SIRT1 and PGC-1α (P<0.05), but had no interaction on mRNA expression levels of Caspase-3,Caspase-9 and Bcl-2 (P >0.05). Conclusion Electroacupuncture combined with enriched rehabilitation training can improve antioxidant capacity,alleviate nerve cell injury,and exert a neuroprotective effect by regulating the SIRT1/PGC-1α pathway.

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姚晓雯,唐 巍,李梦醒,兰 崴,王 玉,高云云,金子开.基于SIRT1/PGC-1α通路探讨电针联合丰富康复训练对脑缺血大鼠氧化应激的影响[J].安徽中医药大学学报,2021,40(2):47-52

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  • 在线发布日期: 2021-04-14