内皮素-1及其受体在缺血再灌注大鼠脑内的分布特征
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摘要
目的内皮素-1(ET-1)有望成为治疗脑缺血性卒中的新靶点,且卒中后其血浆含量增多,但脑内变化情况尚不清楚。文章主要研究ET-1及其受体(ETR)在大鼠脑缺血卒中后各脑区的表达量和分布特征,以推断其与相关脑损伤的关系。方法将12只健康雄性SD大鼠,随机数字表法分为假手术组和缺血再灌注组,每组6只。两组均建立大脑中动脉闭塞/再灌注模型,、但假手术组LICA不插入线栓。免疫组织化学法观测ET-1、ETR的脑区分布特征。结果缺血再灌注组梗死半球皮层中、尾壳核和海马中ET-1表达(0.230±0.019、0.198±0.008、0.183±0.002)较假手术组(0.107±0.005、0.110±0.028、0.101±0.005)明显升高(P<0.05);缺血再灌注组梗死半球脑室脉络丛、软脑膜血管、大脑中动脉血管中A型受体(ETRA)表达(0.131±0.001、0.132±0.005、0.121±0.008)较假手术组(0.086±0.009、0.063±0.003、0.079±0.003)明显升高(P<0.05);缺血再灌注组梗死半球皮层及海马中B型受体(ETRB)的表达(0.187±0.025、0.226±0.019)较假手术组(0.032±0.003、0.029±0.002)明显升高(P<0.05)。结论脑缺血再灌注后,脑内ET-1、ETR于各脑区内呈选择性表达,且其分布位点与缺血再灌注中受损的脑区可能相关。
Objective Endothelin-1(ET-1)is expected to be a new therapeutic target for cerebral ischemic stroke(CIS).The content of plasma is known to increase after stroke,but the changes in the brain remain unclear. The aim of this study was to investigate the expression and distribution of ET-1 and endothelin receptors(ETR)in the brain after CIS in rats and their relationship with related brain injury.Methods We established a model of middle cerebral artery occlusion(MCAO)-reperfusion in 12 male SD rats,6 with nylon filament inserted through the left internal carotid artery(the ischemia-reperfusion[IR]group)and the other 6 without(the sham operation group). We observed the distribution of ET-1 and ETRs in different regions of the brain by immunohistochemistry.Results Compared with the sham operation group,the IR group showed significantly higher expressions of ET-1 in the cortex(0.107 ± 0.005 vs 0.230 ± 0.019,P < 0.05),caudate putamen(0.110 ± 0.028 vs 0.198 ± 0.008,P < 0.05) and hippocampus(0.101 ± 0.005 vs 0.183 ± 0.002,P < 0.05),ETR-A in the bilateral choroid plexus(0.086 ± 0.009 vs 0.131 ± 0.001,P < 0.05),pial vessels(0.063 ± 0.003 vs 0.132 ± 0.005,P < 0.05)and middle cerebral arteries(0.079 ± 0.003 vs 0.121 ± 0.008,P < 0.05),and ETR-B in the infarcted cortex(0.032 ± 0.003 vs 0.187 ± 0.025,P < 0.05)and hippocampus(0.029 ± 0.002 vs 0.226 ± 0.019,P < 0.05)of the ischemic hemisphere.Conclusion After cerebral IR,ET-1 and ETRs are selectively expressed in different brain regions,and their distribution may be related to IR-induced brain injury.
Objective Endothelin-1(ET-1)is expected to be a new therapeutic target for cerebral ischemic stroke(CIS).The content of plasma is known to increase after stroke,but the changes in the brain remain unclear. The aim of this study was to investigate the expression and distribution of ET-1 and endothelin receptors(ETR)in the brain after CIS in rats and their relationship with related brain injury.Methods We established a model of middle cerebral artery occlusion(MCAO)-reperfusion in 12 male SD rats,6 with nylon filament inserted through the left internal carotid artery(the ischemia-reperfusion[IR]group)and the other 6 without(the sham operation group). We observed the distribution of ET-1 and ETRs in different regions of the brain by immunohistochemistry.Results Compared with the sham operation group,the IR group showed significantly higher expressions of ET-1 in the cortex(0.107 ± 0.005 vs 0.230 ± 0.019,P < 0.05),caudate putamen(0.110 ± 0.028 vs 0.198 ± 0.008,P < 0.05) and hippocampus(0.101 ± 0.005 vs 0.183 ± 0.002,P < 0.05),ETR-A in the bilateral choroid plexus(0.086 ± 0.009 vs 0.131 ± 0.001,P < 0.05),pial vessels(0.063 ± 0.003 vs 0.132 ± 0.005,P < 0.05)and middle cerebral arteries(0.079 ± 0.003 vs 0.121 ± 0.008,P < 0.05),and ETR-B in the infarcted cortex(0.032 ± 0.003 vs 0.187 ± 0.025,P < 0.05)and hippocampus(0.029 ± 0.002 vs 0.226 ± 0.019,P < 0.05)of the ischemic hemisphere.Conclusion After cerebral IR,ET-1 and ETRs are selectively expressed in different brain regions,and their distribution may be related to IR-induced brain injury.
引文
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[6] Palomares M,Sara MJC. Myogenic Tone as a Therapeutic Target for Ischemic Stroke[J]Curr Vasc Pharmacol,2013,12(6):1-13.
[7] Kaundal RK,Deshpande TA,Gulati A,et al. Targeting endothelin receptors for pharmacotherapy of ischemic stroke:current scenario and future perspectives[J]. Drug Discov Today,2012,17(13-14):793-804.
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[11] Matsuo Y,Mihara SI,Ninomiya M,et al. Protective Effect of Endothelin Type A Receptor Antagonist on Brain Edema and Injury After Transient Middle Cerebral Artery Occlusion in Rats[J]. Stroke,2001,32(9):2143-2148.
[12] Duan J,Cui J,Yang Z,et al. Neuroprotective effect of Apelin13 on ischemic stroke by activating AMPK/GSK-3β/Nrf2 signaling[J]. J Neuroinflammation,2019,16(1):24.
[13] Chouchani ET,Pell VR,Gaude E,et al. Ischaemic accumulation of succinate controls reperfusion injury through mitochondrial ROS[J]. Nature,2014,515(7527):431-435.
[14] Briyal S,Gulati A. Endothelin-A receptor antagonist BQ123 potentiates acetaminophen induced hypothermia and reduces infarction following focal cerebral ischemia in rats[J]. Eur J Pharmacol,2010,644(1-3):73-79.
[15] Sun M,Izumi H,Shinoda Y,et al. Neuroprotective effects of protein tyrosine phosphatase 1B inhibitor on cerebral ischemia/reperfusion in mice[J]. Brain Res,2018,1694:1-12.
[16]石瑞峰,刘玲,胡斌,等.组织激肽释放酶对糖尿病大鼠局灶性脑缺血再灌注后炎症损伤的干预作用[J].医学研究生学报,2015,28(9):904-909.
[17]段立辉,张丽,刘新峰.脑缺血-再灌注损伤相关的免疫细胞因子[J].医学研究生学报,2004,17(7):651-653.
[18] Hostenbach S,D haeseleer M,Kooijman R,et al. The pathophysiological role of astrocytic endothelin-1[J]. Progress in Neurobiology,2016,144:88-102.
[19] Globus MY,Busto R,Martinez E,et al. Comparative Effect of Transient Global Ischemia on Extracellular Levels of Glutamate,Glycine,andγ-Aminobutyric Acid in Vulnerable and Nonvulnerable Brain Regions in the Rat[J]. J Neurochem,2010,57(2):470-478.
[20] Pacheco-Quinto J,Eckman CB,Eckman EA. Major amyloid-?2-degrading enzymes,endothelin-converting enzyme-2 and neprilysin,are expressed by distinct populations of GABAergic interneurons in hippocampus and neocortex[J]. Neurobio Aging,2016,48:83-92.
[21] Andresen J,Shafi NI,Bryan RM Jr. Endothelial influences on cerebrovascular tone[J]. J Appl Physiol,2006,100(1):318-327.
[22] Palomares SM,Cipolla MJ. Vascular Protection Following Cerebral Ischemia and Reperfusion[J]. J Neurol Neurophysiol,2011,2011(1):1-25.
[23] Zhang Y,Belayev L,Zhao W,et al. A selective endothelin ETA receptor antagonist,SB 234551,improves cerebral perfusion following permanent focal cerebral ischemia in rats[J].Brain Res,2005,1045(1-2):150-156.
[24] Cifuentes EG,Hornick MG,Havalad S,et al. Neuroprotective Effect of IRL-1620,an Endothelin B Receptor Agonist,on a Pediatric Rat Model of Middle Cerebral Artery Occlusion[J]. Front Pediatr 2018,2018(6):1-9.
[2]张胜行,陶静,陈立典.电针治疗对缺血再灌注损伤大鼠Nrg-1/ErbB4信号通路抑制细胞凋亡的影响[J]医学研究生学报,2018,31(12):1246-1253.
[3] Ducci RD,Lange MC,Zétola VHF. Predictors of in-hospital mortality and dependence at discharge in patients with MCA stroke with intravenous thrombolysis[J] Intern Emerg Med,2017,12(4):453-460.
[4] Liu L,Wang D,Wong KSL,et al. Stroke and Stroke Care in China:Huge Burden,Significant Workload,and a National Priority[J]Stroke,2011,42(12):3651-3654.
[5] Davenport AP,Hyndman KA,Dhaun N,et al. Endothelin[J].Pharmacol Rev,2016,68(2):357-418.
[6] Palomares M,Sara MJC. Myogenic Tone as a Therapeutic Target for Ischemic Stroke[J]Curr Vasc Pharmacol,2013,12(6):1-13.
[7] Kaundal RK,Deshpande TA,Gulati A,et al. Targeting endothelin receptors for pharmacotherapy of ischemic stroke:current scenario and future perspectives[J]. Drug Discov Today,2012,17(13-14):793-804.
[8] Moldes O,Sobrino T,Blanco M,et al. Neuroprotection afforded by antagonists of endothelin-1 receptors in experimental stroke[J]. Neuropharmacology,2012,63(8):1279-1285.
[9] Michinaga S,Kimura A,Hatanaka S,et al. Delayed administration of BQ788,an ETB antagonist,after experimental traumatic brain injury promotes recovery of blood-brain barrier function and reduction of cerebral edema in mice[J]. J Neurotrauma,2018,35(13):1481-1494.
[10] Sommer CJ. Ischemic stroke:experimental models and reality[J]. Acta Neuropathol,2017,133(2):245-261.
[11] Matsuo Y,Mihara SI,Ninomiya M,et al. Protective Effect of Endothelin Type A Receptor Antagonist on Brain Edema and Injury After Transient Middle Cerebral Artery Occlusion in Rats[J]. Stroke,2001,32(9):2143-2148.
[12] Duan J,Cui J,Yang Z,et al. Neuroprotective effect of Apelin13 on ischemic stroke by activating AMPK/GSK-3β/Nrf2 signaling[J]. J Neuroinflammation,2019,16(1):24.
[13] Chouchani ET,Pell VR,Gaude E,et al. Ischaemic accumulation of succinate controls reperfusion injury through mitochondrial ROS[J]. Nature,2014,515(7527):431-435.
[14] Briyal S,Gulati A. Endothelin-A receptor antagonist BQ123 potentiates acetaminophen induced hypothermia and reduces infarction following focal cerebral ischemia in rats[J]. Eur J Pharmacol,2010,644(1-3):73-79.
[15] Sun M,Izumi H,Shinoda Y,et al. Neuroprotective effects of protein tyrosine phosphatase 1B inhibitor on cerebral ischemia/reperfusion in mice[J]. Brain Res,2018,1694:1-12.
[16]石瑞峰,刘玲,胡斌,等.组织激肽释放酶对糖尿病大鼠局灶性脑缺血再灌注后炎症损伤的干预作用[J].医学研究生学报,2015,28(9):904-909.
[17]段立辉,张丽,刘新峰.脑缺血-再灌注损伤相关的免疫细胞因子[J].医学研究生学报,2004,17(7):651-653.
[18] Hostenbach S,D haeseleer M,Kooijman R,et al. The pathophysiological role of astrocytic endothelin-1[J]. Progress in Neurobiology,2016,144:88-102.
[19] Globus MY,Busto R,Martinez E,et al. Comparative Effect of Transient Global Ischemia on Extracellular Levels of Glutamate,Glycine,andγ-Aminobutyric Acid in Vulnerable and Nonvulnerable Brain Regions in the Rat[J]. J Neurochem,2010,57(2):470-478.
[20] Pacheco-Quinto J,Eckman CB,Eckman EA. Major amyloid-?2-degrading enzymes,endothelin-converting enzyme-2 and neprilysin,are expressed by distinct populations of GABAergic interneurons in hippocampus and neocortex[J]. Neurobio Aging,2016,48:83-92.
[21] Andresen J,Shafi NI,Bryan RM Jr. Endothelial influences on cerebrovascular tone[J]. J Appl Physiol,2006,100(1):318-327.
[22] Palomares SM,Cipolla MJ. Vascular Protection Following Cerebral Ischemia and Reperfusion[J]. J Neurol Neurophysiol,2011,2011(1):1-25.
[23] Zhang Y,Belayev L,Zhao W,et al. A selective endothelin ETA receptor antagonist,SB 234551,improves cerebral perfusion following permanent focal cerebral ischemia in rats[J].Brain Res,2005,1045(1-2):150-156.
[24] Cifuentes EG,Hornick MG,Havalad S,et al. Neuroprotective Effect of IRL-1620,an Endothelin B Receptor Agonist,on a Pediatric Rat Model of Middle Cerebral Artery Occlusion[J]. Front Pediatr 2018,2018(6):1-9.