三亚市中医院针灸科,海南 三亚 572000
R-33
Department of Acupuncture and Moxibustion, Sanya Hospital of Traditional Chinese Medicine, Sanya 572000
目的 探究电针对局灶性脑缺血大鼠突触可塑性及小胶质细胞极化,并检测 miR-21 / 信号转导和转录激活因子 3(STAT3)通路表达情况。 方法 颈外动脉远端用细线双重结扎 60 min 构建局灶性脑缺血大鼠模型, 神经功能缺损评分评价模型制备情况,模型成功大鼠分为模型组、电针组、尼莫地平组,同时假手术组对照处理。 治疗 3 周结束后神经功能缺损评分评估各组大鼠脑损伤程度;2, 3, 5-三苯基氯化四氮唑(TTC)染色测定各组大鼠脑梗死体积;电镜观察脑皮层突触超微结构并行形态计量学分析;免疫荧光染色观察脑皮层小胶质细胞极化情况;实时荧光定量 PCR( qRT-PCR) 法测定各组大鼠脑皮层中 miR-21 水平;蛋白印迹法测定各组大鼠脑皮层中 Janus 蛋白酪氨酸激酶 2(JAK2)、p-JAK2、STAT3、p-STAT3 水平。 结果 与假手术组相比,模型组大鼠神经功能缺损评分、梗死体积、脑皮层区 M1 型小胶质细胞数量、miR-21、p-STAT3、p-JAK2 水平显著升高(P<0. 05),脑皮层区 M2 型小胶质细胞数量、突触面数密度(Nv)、突触体密度(Vv)、突触连接带面密度( Sv )、突触后致密物质(PSD)、突触界面曲面率、突触间隙宽度明显降低(P<0. 05);与模型组相比,电针组和尼莫地平组大鼠神经功能缺损评分、脑梗死体积、M1 型小胶质细胞数量、miR-21、p-STAT3、p-JAK2 水平显著降低(P<0. 05),M2 型小胶质细胞数量、Nv、 Vv、Sv、PSD、突触界面曲面率、突触间隙宽度显著升高(P<0. 05)。 结论 电针可促进局灶性脑缺血大鼠突触重建, 诱导小胶质细胞由 M1 向 M2 型极化。
Objective To explore the effects of electroacupuncture on synaptic plasticity and microglial polarization and detect the expression of miR-21 / signal transducer and activator of transcription 3 (STAT3) pathway in rats with cerebral ischemia. Methods The distal end of the external carotid artery was double-ligated with a thin wire for 60 minutes to establish a rat model of focal cerebral ischemia. The neurological deficit score was used to evaluate the model preparation. The successful model rats were divided into a model group, electroacupuncture group, and nimodipine group.The control group comprised rats treated by a sham operation. After 3 weeks of treatment, the cerebral infarction volume was measured by 2,3,5-triphenyltetrazolium chloride staining, the synapse ultrastructure was observed by electron microscopy and analyzed by morphometry, and the microglial polarization was assessed by immunofluorescence staining. Additionally, the level of miR-21 in the cerebral cortex of rats in each group was measured by real-time fluorescence quantitative polymerase chain reaction, and western blotting was used to detect the levels of Janus kinase 2 (JAK2), p-JAK2, STAT3, and p-STAT3 in the cerebral cortex of rats in each group. Results The neurological deficit score, cerebral infarction volume, number of M1 microglia, and levels of miR-21, p-STAT3, and p-JAK2 were significantly higher and the number of M2 microglia, numerical density of synapsis (Nv ), volume density of synapsis (Vv ), surface density of synapsis ( Sv ), postsynaptic density (PSD), synaptic curvature, and width of the synaptic cleft were significantly lower in the model group than in the control group (P< 0. 05). The neurological deficit score, cerebral infarction volume, number of M1 microglia, and levels of miR-21, p-STAT3, and p-JAK2 were significantly lower and the number of M2 microglia, Nv, Vv, Sv, PSD, synaptic curvature, and width of the synaptic cleft were significantly higher in the electroacupuncture group and nimodipine group than in the model group (P<0. 05). Conclusions Electroacupuncture can promote synaptic remodeling and induce microglial polarization from M1 to M2 in rats with focal cerebral ischemia.
徐 琼,刘建浩,王天磊,谭春凤.电针对脑缺血大鼠突触可塑性及小胶质细胞极化的影响[J].中国比较医学杂志,2021,31(6):48~55.
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