电针对局灶性脑缺血大鼠缺血皮质区内皮抑素、血小板反应蛋白-1表达的影响
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摘要
目的观察电针对局灶性脑缺血大鼠缺血皮质区内皮抑素(endostatin,ES)、血小板反应蛋白-1(thrombospondin-1,TSP-1)表达的调控作用及其机制。方法将54只健康雄性SD大鼠随机分为假手术组、模型组和电针组,每组18只。利用改良线栓法复制局灶性大脑中动脉闭塞模型,电针组选取"百会"与"水沟"穴进行治疗,每次30min,每日1次,共干预7d。比较术后7d各组大鼠神经功能评分,用2,3,5-三苯基氯化四氮唑染色法测量脑梗死面积,免疫组织化学法及蛋白免疫印迹法检测ES、TSP-1蛋白在大鼠脑缺血皮质区的表达情况。结果与假手术组比较,模型组大鼠神经功能缺损体征较为显著,梗死范围明显,脑缺血皮质区ES和TSP-1蛋白表达均显著上调(P<0.05);与模型组比较,电针组大鼠神经功能评分降低(P<0.05),梗死面积缩小(P<0.05),脑缺血皮质区ES、TSP-1蛋白表达水平均明显下调(P<0.05)。结论电针可促进局灶性脑缺血大鼠神经功能恢复,缩小梗死面积,其机制可能与血管新生抑制因子ES、TSP-1的表达下调有关。
Objective To observe the effect of electroacupuncture(EA)on the expression of endostatin(ES)and thrombospondin-1(TSP-1)in the ischemic cortex in rats with focal cerebral ischemia(FCI)and its mechanism.Methods Fifty-four healthy male Sprague-Dawley rats were randomly divided into shamoperation group,model group,and EA group,with 18 rats in each group.A right middle cerebral artery occlusion model was established by the modified suture method.The EA group was treated by EA(30 min)at Baihui and Shuigou points once daily for 7 days.The neurological score was assessed at 7 dafter operation.The cerebral infarct area was measured by 2,3,5-triphenyltetrazolium chloride staining.Immunohistochemistry and Western blot were used to measure the protein expression of ES and TSP-1 in the ischemic cortex.Results Compared with the sham-operation group,the model group showed obvious signs of neurological deficit,with a significantly increased neurological score,an obvious infarct area,and significantly upregulated protein expression of ES and TSP-1 in the ischemic cortex(P<0.05).Compared with the model group,the EA group had a significantly lower neurological score(P<0.05),a significantly smaller infarct area(P<0.05),and significantly downregulated expression of ES and TSP-1 in the ischemic cortex(P<0.05).Conclusion EA can promote neurological recovery and reduce the infarct area in rats with FCI,possibly by down-regulating the expression of angiogenesis inhibitors ES and TSP-1.
Objective To observe the effect of electroacupuncture(EA)on the expression of endostatin(ES)and thrombospondin-1(TSP-1)in the ischemic cortex in rats with focal cerebral ischemia(FCI)and its mechanism.Methods Fifty-four healthy male Sprague-Dawley rats were randomly divided into shamoperation group,model group,and EA group,with 18 rats in each group.A right middle cerebral artery occlusion model was established by the modified suture method.The EA group was treated by EA(30 min)at Baihui and Shuigou points once daily for 7 days.The neurological score was assessed at 7 dafter operation.The cerebral infarct area was measured by 2,3,5-triphenyltetrazolium chloride staining.Immunohistochemistry and Western blot were used to measure the protein expression of ES and TSP-1 in the ischemic cortex.Results Compared with the sham-operation group,the model group showed obvious signs of neurological deficit,with a significantly increased neurological score,an obvious infarct area,and significantly upregulated protein expression of ES and TSP-1 in the ischemic cortex(P<0.05).Compared with the model group,the EA group had a significantly lower neurological score(P<0.05),a significantly smaller infarct area(P<0.05),and significantly downregulated expression of ES and TSP-1 in the ischemic cortex(P<0.05).Conclusion EA can promote neurological recovery and reduce the infarct area in rats with FCI,possibly by down-regulating the expression of angiogenesis inhibitors ES and TSP-1.
引文
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[14]王琼,黄伟,吴洪阳,等.针刺百会、人中穴对急性脑缺血大鼠模型葡萄糖转运蛋白1、3的影响[J].广州中医药大学学报,2018,35(1):86-92.
[15]杨敏,陈邦国,梁超.头针对大鼠局灶性脑缺血再灌注后脑组织VEGF及ES表达的影响[J].中国中医急症,2014,23(6):1012-1013,1052.
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[18]MAATTA M,HELJASVAARA R,PIHLAJANIE-MI T,et al.Collagen XVIII/endostatin shows a ubiquitous distribution in human ocular tissues and endostain-containing fragments accumulate in ocular fluid samples[J].Graefes Arch Clin Exp Ophthalmol,2006,25(4):341-349.
[19]马曦,罗勇.电针对大鼠局灶脑缺血再灌注后脑内血管生长因子和血管抑制因子表达的影响[J].中国针灸,2006,27(2):129-133.
[20]TIAN H L,CHEN H,CUI Y H,et al.Increased protein and mRNA expression of endostatin in the ischemic brain tissue of rabbits after middle cerebral artery occlusion[J].Neurosci Bull,2007,23(1):35-40.
[21]SHIRAHA H,YAMAMOTO K,NAMBA M.Human hepatocyte carcinogenesis(Review)[J].Int J Oncol,2013,42(4):1133-1138.
[22]LEE D,KIM D,CHOI Y B,et al.Simultaneous blockade of VEGF and Dll4by HD105,a bispecific antibody,inhibits tumor progression and angiogenesis[J].MAbs,2016,8(5):892-904.
[23]孙顺昌,王国峰,赵玉芳,等.局灶性缺血预处理对脑缺血大鼠血管内皮生长因子表达及血管形成的影响[J].中国临床神经科学,2011,19(6):594-600.
[24]蒋静子,刘开祥,李浩,等.急性脑梗死患者血清VEGF与内皮抑素水平的动态变化及意义[J].广西医科大学学报,2012,29(6):869-871.
[25]万赛英,谭峰,吴海科,等.电针干预高血压大鼠脑缺血Ang1TSP-1的表达及细胞超微结构的研究[J].中华中医药学刊,2010,28(12):2544-2547.
[2]CUNNINGHAM C J,REDONDO-CASTRO E,ALLANS M.The therapeutic potential of the mesenchymal stem cell secretome in ischaemic stroke[J].J Cereb Blood Flow Metab,2018,38(8):1276-1292.
[3]MURRAY C J,LOPEZ A D.Measuring the global burden of disease[J].N Engl J Med,2013,369(5):448-457.
[4]MA J X,ZHANG L N,NIU T F,et al.Growth differentiation factor 11improves neurobehavioral recovery and stimulates angiogenesis in rats subjected to cerebral ischemia/reperfusion[J].Brain research bulletin,2018,139:38-47.
[5]SHI L,CAO H M,LI Y,et al.Electroacupuncture improves neurovascular unit reconstruction by promoting collateral circulation and angiogenesis[J].Neural regeneration research,2017,12(12):2000-2006.
[6]XIE M,YI C,LUO X,et al.Glial gap junctional communication involvement in hippocampal damage after middle cerebral artery occlusion[J].Ann Neurol,2011(70):121-132.
[7]姚红,倪光夏.基于“治疗性血管新生”理论探讨针灸治疗缺血性脑卒中的机制[J].吉林中医药,2016,36(5):521-525.
[8]郭义.实验针灸学[M].北京:中国中医药出版社,2008,14(7):414-417.
[9]LONGA F Z,WEINSTEIN P R,CARLSON S,et al.Reversible middle cerebral artery occlusion without craniectomy in rats[J].Stroke,1989,20(1):84.
[10]罗勇,董为伟.Wistar大鼠插线法局灶性脑缺血/再灌注模型的实验研究[J].重庆医科大学学报,2002(1):1-4.
[11]BEDERSON J B,PITTS L H,TSUJI M,et al.Rat middle cerebral artery occlusion evaluation of model and development of aneurologic examination[J].Stroke,1986,17(3):472-476.
[12]王琼,黄伟,吴洪阳.针刺对急性脑缺血大鼠沉默信息调节因子2相关酶1和核转录因子-κB蛋白的影响[J].针刺研究,2018,43(3):146-151.
[13]黄伟,李佳,朱广为.针刺百会、人中穴对急性脑缺血大鼠模型NF-κB/IκB-α的影响[J].中华中医药杂志,2017,32(1):298-302.
[14]王琼,黄伟,吴洪阳,等.针刺百会、人中穴对急性脑缺血大鼠模型葡萄糖转运蛋白1、3的影响[J].广州中医药大学学报,2018,35(1):86-92.
[15]杨敏,陈邦国,梁超.头针对大鼠局灶性脑缺血再灌注后脑组织VEGF及ES表达的影响[J].中国中医急症,2014,23(6):1012-1013,1052.
[16]PALMER T D,WILLHOITE A R,GAGE F H.Vascular niche for adult hippocampal neurogenesis[J].J Comp Neurol,2000,425(4):479-494.
[17]SU S J,YEH T M,CHUANG W J,et al.The novel targets for anti-angiogenesis of genistein on human cancer cells[J].Biochem Pharmacol,2005,69(2):307.
[18]MAATTA M,HELJASVAARA R,PIHLAJANIE-MI T,et al.Collagen XVIII/endostatin shows a ubiquitous distribution in human ocular tissues and endostain-containing fragments accumulate in ocular fluid samples[J].Graefes Arch Clin Exp Ophthalmol,2006,25(4):341-349.
[19]马曦,罗勇.电针对大鼠局灶脑缺血再灌注后脑内血管生长因子和血管抑制因子表达的影响[J].中国针灸,2006,27(2):129-133.
[20]TIAN H L,CHEN H,CUI Y H,et al.Increased protein and mRNA expression of endostatin in the ischemic brain tissue of rabbits after middle cerebral artery occlusion[J].Neurosci Bull,2007,23(1):35-40.
[21]SHIRAHA H,YAMAMOTO K,NAMBA M.Human hepatocyte carcinogenesis(Review)[J].Int J Oncol,2013,42(4):1133-1138.
[22]LEE D,KIM D,CHOI Y B,et al.Simultaneous blockade of VEGF and Dll4by HD105,a bispecific antibody,inhibits tumor progression and angiogenesis[J].MAbs,2016,8(5):892-904.
[23]孙顺昌,王国峰,赵玉芳,等.局灶性缺血预处理对脑缺血大鼠血管内皮生长因子表达及血管形成的影响[J].中国临床神经科学,2011,19(6):594-600.
[24]蒋静子,刘开祥,李浩,等.急性脑梗死患者血清VEGF与内皮抑素水平的动态变化及意义[J].广西医科大学学报,2012,29(6):869-871.
[25]万赛英,谭峰,吴海科,等.电针干预高血压大鼠脑缺血Ang1TSP-1的表达及细胞超微结构的研究[J].中华中医药学刊,2010,28(12):2544-2547.