迷走神经刺激通过自噬机制对短暂性大脑中动脉栓塞大鼠模型神经保护作用的实验研究
|
摘要
目的探讨迷走神经刺激对大鼠短暂性大脑中动脉栓塞(t-MCAO)模型的神经保护作用及其可能的机制。方法将雄性SD大鼠60只,随机平分为3组,即迷走神经刺激组(VNS组,造模后予迷走神经刺激)、模型组(重复VNS组步骤但不刺激迷走神经)及假手术组(重复VNS组步骤,但不栓塞大脑中动脉及刺激迷走神经)。其中,采用线栓法将大脑中动脉闭塞,术后2 h拔出栓线,建立t-MCAO模型。各组给予相应操作流程后24 h,记录神经行为学评分、计算脑梗死体积及观察脑皮质区LC3-Ⅱ、Beclin1的表达。结果模型组神经行为学评分、脑梗死体积较假手术组均显著恶化(P <0. 05),VNS组神经行为学评分、脑梗死体积较模型组有改善(P <0. 05),但仍差于假手术组(P<0. 05)。相对于假手术组,模型组LC3-Ⅱ、Beclin1表达显著升高(P <0. 05),VNS组LC3-Ⅱ、Beclin1表达水平相对于模型组显著降低(P <0. 05),但仍高于假手术组(P <0. 05)。结论迷走神经刺激可能通过调控自噬激活水平发挥对大鼠短暂性脑缺血损伤的神经保护作用。
Objective To investigate the neuroprotective effects and mechanism of vagal nerve stimulation( VNS) in a rat transient middle cerebral artery occlusion model. Methods 60 male SD rats were randomly divided into three groups: VNS group( n = 20),model group( n= 20),SHAM operation group( n = 20). The model of rat transient focal cerebral ischemia was induced by the intraluminal suture method. The model group repeated the steps of the VNS-treated group,but did not stimulate. The sham operation group repeated the experimental steps,but it neither embolized the vessels nor stimulated nerves. The rats were sacrificed after 24 h. Indices of neurobehavioral scores,infarction rate were measured. The expressions of LC3-Ⅱand Beclin1 in brain tissue were determined by immunohistochemistry staining. Results Compared with the sham operation group,Indices of neurobehavioral scores,infarction rate were significantly improved( P < 0. 05). There were also significant differences between VNS group and model group( P < 0. 05). Compared with model group and sham operation group,the number of positive cells of LC3-Ⅱand Beclin1 in the VNS-treated group decreased significantly( P < 0. 05). Compared with the sham operation group,it were significantly improved in Model group( P < 0. 05). Conclusion The neuroprotective mechanism of VNS for cerebral ischemia may be associated with the inhibition of autophagy.
Objective To investigate the neuroprotective effects and mechanism of vagal nerve stimulation( VNS) in a rat transient middle cerebral artery occlusion model. Methods 60 male SD rats were randomly divided into three groups: VNS group( n = 20),model group( n= 20),SHAM operation group( n = 20). The model of rat transient focal cerebral ischemia was induced by the intraluminal suture method. The model group repeated the steps of the VNS-treated group,but did not stimulate. The sham operation group repeated the experimental steps,but it neither embolized the vessels nor stimulated nerves. The rats were sacrificed after 24 h. Indices of neurobehavioral scores,infarction rate were measured. The expressions of LC3-Ⅱand Beclin1 in brain tissue were determined by immunohistochemistry staining. Results Compared with the sham operation group,Indices of neurobehavioral scores,infarction rate were significantly improved( P < 0. 05). There were also significant differences between VNS group and model group( P < 0. 05). Compared with model group and sham operation group,the number of positive cells of LC3-Ⅱand Beclin1 in the VNS-treated group decreased significantly( P < 0. 05). Compared with the sham operation group,it were significantly improved in Model group( P < 0. 05). Conclusion The neuroprotective mechanism of VNS for cerebral ischemia may be associated with the inhibition of autophagy.
引文
[1] Miyamoto O,Pang J,Sumitani K,et al. Mechanisms of the anti-ischemic effect of vagus nerve stimulation in the gerbil hippocampus[J].Neuroreport,2003,14(15):1971-1974.
[2] Sun Z,Baker W,Hiraki T,et al. The effect of right vagus nerve stimulation on focal cerebral ischemia:an experimental study in the rat[J].Brain Stimulation,2012,5(1):1-10.
[3] Ay I,Lu J,Ay H,et al. Vagus nerve stimulation reduces infarct size in rat focal cerebral ischemia[J]. Neurosci Lett,2009,459(3):147-151.
[4] Levine B,Kroemer G. Autophagy in the pathogenesis of disease[J].Cell,2008,132(1):27-42.
[5]王辉,王蓉,何娅,等.大鼠脑梗死模型神经行为学评价的可靠性探讨[J].陕西医学杂志,2015,44(6):649-651.
[6] Chiang T,Messing RO,Chou WH. Mouse model of middle cerebral artery occlusion[J]. J Vis Exp,2011,(48).
[7] Smith DC,Modglin AA,Roosevelt RW,et al. Electrical Stimulation of the Vagus Nerve Enhances Cognitive and Motor Recovery following Moderate Fluid Percussion Injury in the Rat[J]. J Neurotrauma,2005,22(12):1485-1502.
[8] Groves DA,Brown VJ. Vagal nerve stimulation:a review of its applications and potential mechanisms that mediate its clinical effects[J]. Neurosci Biobehav Rev,2005,29(3):493-500.
[9] Bonaz B,Picq C,Sinniger V,et al. Vagus nerve stimulation:from epilepsy to the cholinergic anti-inflammatory pathway[J]. Neurogastroenterol Motil,2013,25(3):208-221.
[10] Dorr AE,Debonnel G. Effect of Vagus Nerve Stimulation on Serotonergic and Noradrenergic Transmission[J]. J Pharmacol Exp Ther,2006,318(2):890-898.
[11] Cheyuo C,Jacob A,Wu R,et al. The parasympathetic nervous system in the quest for stroke therapeutics[J]. J Cereb Blood Flow Metab,2011,31(5):1187-1195.
[12] Moskowitz MA,Lo EH,Iadecola C. The science of stroke:mechanisms in search of treatments[J]. Neuron,2010,67(2):181-198.
[13]王忠,孙建新,连军,等.迷走神经阻断对脑缺血大鼠模型的影响[J].新疆医科大学学报,2013,36(8):1105-1107.
[14]乐南阳,肖波,马延斌.植入性迷走神经刺激治疗顽固性癫痫的研究进展[J].中华神经外科杂志,2017,33(3):318-320.
[15] Kalinin S,Polak PE,Madrigal JL,et al. Beta-amyloid-dependent expression of NOS2 in neurons:prevention by an alpha2-adrenergic antagonist[J]. Antioxid Redox Signal,2006,8(5-6):873-883.
[16] Shytle RD,Mori T,Townsend K,et al. Cholinergic modulation of microglial activation by alpha 7 nicotinic receptors[J]. J Neurochem,2004,89(2):337-343.
[17]田云丽,施静,刘爱芬.迷走神经刺激术在脑缺血中的应用及神经机制[J].中国组织化学与细胞化学杂志,2012,21(3):312-315.
[18]叶樱泽,熊晓星.自噬与脑卒中的研究进展[J].卒中与神经疾病,2018,25(3):326-328,347.
[19] Tian F,Deguchi K,Yamashita T,et al. In vivo imaging of autophagy in a mouse stroke model[J]. Autophagy,2010,6(8):1107-1114.
[20] Wen YD,Sheng R,Zhang LS,et al. Neuronal injury in rat model of permanent focal cerebral ischemia is associated with activation of autophagic and lysosomal pathways[J]. Autophagy,2008,4(6):762-769.
[21] Carloni S,Buonocore G,Balduini W. Protective role of autophagy in neonatal hypoxia-ischemia induced brain injury[J]. Neurobiol Dis,2008,32(3):329-339.
[22] Lu Y,Kang J,Bai Y,et al. Hyperbaric oxygen enlarges the area of brain damage in MCAO rats by blocking autophagy via ERK1/2 activation[J]. Eur J Pharmacol,2014,728:93-99.
[23] Zhang X,Shen Y,Chen Z,et al. Endoplasmic reticulum stress induced by tunicamycin and thapsigargin protects against transient ischemic brain injury:Involvement of PARK2-dependent mitophagy[J]. Autophagy,2014,10(10):1801-1813.
[24] Zheng YQ,Liu JX,Li XZ,et al. RNA interference-mediated downregulation of Beclinl attenuates cerebral ischemic injury in rats[J]. Acta Pharmacol Sin,2009,30(7):919-927.
[25] Zheng C,Han J,Xia W,et al. NAD(+)administration decreases ischemic brain damage partially by blocking autophagy in a mouse model of brain ischemia[J]. Neurosci Lett,2012,512(2):67-71.
[2] Sun Z,Baker W,Hiraki T,et al. The effect of right vagus nerve stimulation on focal cerebral ischemia:an experimental study in the rat[J].Brain Stimulation,2012,5(1):1-10.
[3] Ay I,Lu J,Ay H,et al. Vagus nerve stimulation reduces infarct size in rat focal cerebral ischemia[J]. Neurosci Lett,2009,459(3):147-151.
[4] Levine B,Kroemer G. Autophagy in the pathogenesis of disease[J].Cell,2008,132(1):27-42.
[5]王辉,王蓉,何娅,等.大鼠脑梗死模型神经行为学评价的可靠性探讨[J].陕西医学杂志,2015,44(6):649-651.
[6] Chiang T,Messing RO,Chou WH. Mouse model of middle cerebral artery occlusion[J]. J Vis Exp,2011,(48).
[7] Smith DC,Modglin AA,Roosevelt RW,et al. Electrical Stimulation of the Vagus Nerve Enhances Cognitive and Motor Recovery following Moderate Fluid Percussion Injury in the Rat[J]. J Neurotrauma,2005,22(12):1485-1502.
[8] Groves DA,Brown VJ. Vagal nerve stimulation:a review of its applications and potential mechanisms that mediate its clinical effects[J]. Neurosci Biobehav Rev,2005,29(3):493-500.
[9] Bonaz B,Picq C,Sinniger V,et al. Vagus nerve stimulation:from epilepsy to the cholinergic anti-inflammatory pathway[J]. Neurogastroenterol Motil,2013,25(3):208-221.
[10] Dorr AE,Debonnel G. Effect of Vagus Nerve Stimulation on Serotonergic and Noradrenergic Transmission[J]. J Pharmacol Exp Ther,2006,318(2):890-898.
[11] Cheyuo C,Jacob A,Wu R,et al. The parasympathetic nervous system in the quest for stroke therapeutics[J]. J Cereb Blood Flow Metab,2011,31(5):1187-1195.
[12] Moskowitz MA,Lo EH,Iadecola C. The science of stroke:mechanisms in search of treatments[J]. Neuron,2010,67(2):181-198.
[13]王忠,孙建新,连军,等.迷走神经阻断对脑缺血大鼠模型的影响[J].新疆医科大学学报,2013,36(8):1105-1107.
[14]乐南阳,肖波,马延斌.植入性迷走神经刺激治疗顽固性癫痫的研究进展[J].中华神经外科杂志,2017,33(3):318-320.
[15] Kalinin S,Polak PE,Madrigal JL,et al. Beta-amyloid-dependent expression of NOS2 in neurons:prevention by an alpha2-adrenergic antagonist[J]. Antioxid Redox Signal,2006,8(5-6):873-883.
[16] Shytle RD,Mori T,Townsend K,et al. Cholinergic modulation of microglial activation by alpha 7 nicotinic receptors[J]. J Neurochem,2004,89(2):337-343.
[17]田云丽,施静,刘爱芬.迷走神经刺激术在脑缺血中的应用及神经机制[J].中国组织化学与细胞化学杂志,2012,21(3):312-315.
[18]叶樱泽,熊晓星.自噬与脑卒中的研究进展[J].卒中与神经疾病,2018,25(3):326-328,347.
[19] Tian F,Deguchi K,Yamashita T,et al. In vivo imaging of autophagy in a mouse stroke model[J]. Autophagy,2010,6(8):1107-1114.
[20] Wen YD,Sheng R,Zhang LS,et al. Neuronal injury in rat model of permanent focal cerebral ischemia is associated with activation of autophagic and lysosomal pathways[J]. Autophagy,2008,4(6):762-769.
[21] Carloni S,Buonocore G,Balduini W. Protective role of autophagy in neonatal hypoxia-ischemia induced brain injury[J]. Neurobiol Dis,2008,32(3):329-339.
[22] Lu Y,Kang J,Bai Y,et al. Hyperbaric oxygen enlarges the area of brain damage in MCAO rats by blocking autophagy via ERK1/2 activation[J]. Eur J Pharmacol,2014,728:93-99.
[23] Zhang X,Shen Y,Chen Z,et al. Endoplasmic reticulum stress induced by tunicamycin and thapsigargin protects against transient ischemic brain injury:Involvement of PARK2-dependent mitophagy[J]. Autophagy,2014,10(10):1801-1813.
[24] Zheng YQ,Liu JX,Li XZ,et al. RNA interference-mediated downregulation of Beclinl attenuates cerebral ischemic injury in rats[J]. Acta Pharmacol Sin,2009,30(7):919-927.
[25] Zheng C,Han J,Xia W,et al. NAD(+)administration decreases ischemic brain damage partially by blocking autophagy in a mouse model of brain ischemia[J]. Neurosci Lett,2012,512(2):67-71.