标本配穴电针预处理对脑缺血再灌注损伤大鼠海马沉默信息调节因子1的影响
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摘要
目的基于沉默信息调节因子1(silent information regulation 1, SIRT1)途径探讨标本配穴电针预处理抗脑缺血再灌注损伤的作用机制。方法将60只雄性SD大鼠随机分为A组、B组和C组,每组20只。A组仅分离右侧颈总动脉,不给予其他处理;B组采用改良的MCAO线栓法制备大鼠右侧局灶性脑缺血再灌注损伤模型;C组造模前取百会、肾俞、三阴交行电针预处理。比较各组再灌注3 h后神经行为学评分,并采用TTC染色法测定脑梗死容积百分比,采用HE染色法观察海马神经元形态,采用Western-blot方法检测SIRT1和过氧化物酶体增殖物活化受体γ共激活因子-1α(peroxisome proliferator-activated receptorγcoactivator-1α,PGC-1α)蛋白表达。结果 C组神经行为学评分和脑梗死百分比较B组均显著降低,SIRT1和PGC-1α蛋白表达较B组均显著升高,两组比较差异均具有统计学意义(P<0.01)。结论标本配穴电针预处理可能通过上调内源性脑SIRT1和PGC-1α蛋白表达而减轻脑缺血再灌注损伤。
Objective To explore the action mechanism of electroacupuncture pretreatment with secondary-primary point combination against cerebral ischemia-reperfusion injury by focusing the silent information regulator 1(SIRT1)pathway. Method Sixty male Sprague-Dawley(SD) rats were randomized into group A, group B and group C, with20 rats in each group. Group A only received right common carotid artery separation without any other treatments,while group B was intervened by the modified middle cerebral artery occlusion(MCAO) method to establish the right focal cerebral ischemia-reperfusion injury model, and group C was intervened by electroacupuncture pretreatment at Baihui(GV20), Shenshu(BL23) and Sanyinjiao(SP6) before modeling. Neurobehavioral scores in the three groups were compared three hours after the reperfusion. The percentage of cerebral infarct volume was measured by2,3,5-triphenyltetrazolium chloride(TTC) staining. The morphology of hippocampal neurons was observed by hematoxylin-eosin(HE) staining. The protein expressions of SIRT1 and peroxisome proliferator-activated receptor γcoactivator-1 α(PGC-1α) were examined using Western blotting method. Result Compared to group B, the neurobehavioral score and the percentage of cerebral infarct volume declined significantly in group C, the protein expressions of SIRT1 and PGC-1α both increased significantly in group C, and the between-group differences were statistically significant(P < 0.05). Conclusion Electroacupuncture pretreatment with secondary-primary point combination may alleviate cerebral ischemia-reperfusion injury by upregulating the protein expressions of endogenous SIRT1 and PGC-1α in brain.
Objective To explore the action mechanism of electroacupuncture pretreatment with secondary-primary point combination against cerebral ischemia-reperfusion injury by focusing the silent information regulator 1(SIRT1)pathway. Method Sixty male Sprague-Dawley(SD) rats were randomized into group A, group B and group C, with20 rats in each group. Group A only received right common carotid artery separation without any other treatments,while group B was intervened by the modified middle cerebral artery occlusion(MCAO) method to establish the right focal cerebral ischemia-reperfusion injury model, and group C was intervened by electroacupuncture pretreatment at Baihui(GV20), Shenshu(BL23) and Sanyinjiao(SP6) before modeling. Neurobehavioral scores in the three groups were compared three hours after the reperfusion. The percentage of cerebral infarct volume was measured by2,3,5-triphenyltetrazolium chloride(TTC) staining. The morphology of hippocampal neurons was observed by hematoxylin-eosin(HE) staining. The protein expressions of SIRT1 and peroxisome proliferator-activated receptor γcoactivator-1 α(PGC-1α) were examined using Western blotting method. Result Compared to group B, the neurobehavioral score and the percentage of cerebral infarct volume declined significantly in group C, the protein expressions of SIRT1 and PGC-1α both increased significantly in group C, and the between-group differences were statistically significant(P < 0.05). Conclusion Electroacupuncture pretreatment with secondary-primary point combination may alleviate cerebral ischemia-reperfusion injury by upregulating the protein expressions of endogenous SIRT1 and PGC-1α in brain.
引文
[1]陈泽斌,王华.大鼠穴位针刺预处理抗脑缺血再灌注损伤作用探讨[J].针刺研究,2003,28(3):165-169.
[2]Miao Y,Zhao S,Gao Y,et al.Curcumin pretreatment attenuates inflammation and mitochondrial dysfunction in experimental stroke:The possible role of Sirt1signaling[J].Brain Res Bull,2016,121:9-15.
[3]Yan W,Fang Z,Yang Q,et al.SirT1 mediates hyperbaric oxygen preconditioning-induced ischemic tolerance in rat brain[J].J Cereb Blood Flow Metab,2013,33(3):396-406.
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[7]王华,梁凤霞.腧穴配伍研究思路和展望[J].中国针灸,2012,32(4):359-362.
[8]赵钢,马莹莹,付嘉明.“标本配穴”电针对心肌缺血模型大鼠心脏功能和细胞凋亡指数的影响[J].上海针灸杂志,2018,37(4):461-465.
[9]唐关敏,钱钢,翟昌林,等.针刺预处理对缺血再灌注损伤大鼠心肌HMGB1表达的影响[J].中国中医药科技,2018,25(4):463-466.
[10]黄丽,王春,李言,等.针刺预处理对脑缺血大鼠小脑浦肯野细胞动作电位的影响[J].南方医科大学学报,2018,38(6):677-682.
[11]张松兴,徐月英,邱泽玲.针刺预处理研究进展[J].辽宁中医药大学学报,2007,9(5):72-74.
[12]谢俊,陈泽斌,吴松,等.不同腧穴配伍电针防治大鼠心肌缺血损伤作用的比较[J].针刺研究,2017,42(2):131-135.
[13]徐文博,唐关敏,胡惠林,等.针刺预处理对缺血再灌注损伤大鼠心肌内质网应激的影响[J].中国中医药科技,2018,25(5):620-623.
[14]黄玉栋,聂焱,王志强,等.颅针对脑缺血再灌注损伤大鼠氧化应激的影响[J].上海针灸杂志,2016,35(12):1473-1476.
[15]徐芬,梁凤霞,陈瑞,等.标本配穴电针对胰岛素抵抗大鼠胰岛素敏感性及骨骼肌SIRT1蛋白表达的影响[J].中医杂志,2014,55(4):328-332.
[16]陈泽斌,邹峰,袁芳,等.针刺预处理对脑缺血再灌注大鼠顶皮质NR2A和NR2B蛋白表达的影响[J].中国中西医结合急救杂志,2005,12(2):79-83.
[17]王颖.针刺对脑缺血再灌注模型大鼠ROCK表达影响的动态观察[J].上海针灸杂志,2015,34(4):367-370.
[18]李珩,肖宁,邵明璐,等.针刺内关预处理干预心肌缺血再灌注大鼠实验效应的系统评价[J].中国中医基础医学杂志,2016,22(12):1665-1669.
[19]陈泽斌,柯晖,邹锋.针刺预处理抗脑缺血再灌注损伤作用及其分子机制[J].中医药学刊,2006,24(7):1246-1249.
[20]Paraíso AF,Mendes KL,Santos SH.Brain activation of SIRT1:role in neuropathology[J].Mol Neurobiol,2013,48(3):681-689.
[21]Samuel SM,Thirunavukkarasu M,Penumathsa SV,et al.Akt/FOXO3a/SIRT1-mediated cardioprotection by ntyrosol against ischemic stress in rat in vivo model of myocardial infarction:switching gears toward survival and longevity[J].J Agric Food Chem,2008,56(20):9692-9698.
[22]郭燕世.热量限制与白藜芦醇抗衰老的研究进展[J].生命科学进展,2011,42(3):161-164.
[23]Hernández-Jiménez M,Hurtado O,Cuartero MI,et al.Silent information regulator 1 protects the brain against cerebral ischemic damage[J].Stroke,2013,44(8):2333-2337.
[24]Miao Y,Zhao S,Gao Y,et al.Curcumin pretreatment attenuates inflammation and mitochondrial dysfunction in experimental stroke:the possible role of Sirt1signaling[J].Brain Res Bull,2016,121:9-15.
[25]Chen WK,Tsai YL,Shibu MA,et al.Exercise training augments Sirt1-signaling and attenuates cardiac inflamemation in D-galactose induced-aging rats[J].Aging(Albany NY),2018,10(12):4166-4174.
[26]Gupta AK,Choi S.Integrated Ligand and StructureBased Investigation of Structural Requirements for Silent Information Regulator 1(SIRT1)Activation[J].Curr Top Med Chem,2018,18(27):2313-2324.
[27]CantóC,Auwerx J.PGC-1alpha,SIRTI and AMPK,an energy sensing network that eonirols energy expenditure[J].Curr Opin Lipidol,2009,20(2):98-105.
[28]Wareski P,Vaarmann A,Choubey V,et al.PGC-1{alpha}and PGC-1{beta}regulate mitochondrial density in neurons[J].J Biol Chem,2009,284(32):21379-21385.
[2]Miao Y,Zhao S,Gao Y,et al.Curcumin pretreatment attenuates inflammation and mitochondrial dysfunction in experimental stroke:The possible role of Sirt1signaling[J].Brain Res Bull,2016,121:9-15.
[3]Yan W,Fang Z,Yang Q,et al.SirT1 mediates hyperbaric oxygen preconditioning-induced ischemic tolerance in rat brain[J].J Cereb Blood Flow Metab,2013,33(3):396-406.
[4]Longa EZ,Weinstein PR,Carlson S,et al.Reversible middle cerebral artery occlusion without craniectomy in rats[J].Stroke,1989,20(l):84-91.
[5]华兴邦,李辞蓉,周浩良,等.大鼠穴位图谱的研制[J].实验动物与动物实验,1991,3(1):1-5.
[6]Kato H,Kogure K.Biochemical and molecular characteristics of the brain with developing cerebral infarction[J].Cell Mol Neurobiol,1999,19(1):93-108.
[7]王华,梁凤霞.腧穴配伍研究思路和展望[J].中国针灸,2012,32(4):359-362.
[8]赵钢,马莹莹,付嘉明.“标本配穴”电针对心肌缺血模型大鼠心脏功能和细胞凋亡指数的影响[J].上海针灸杂志,2018,37(4):461-465.
[9]唐关敏,钱钢,翟昌林,等.针刺预处理对缺血再灌注损伤大鼠心肌HMGB1表达的影响[J].中国中医药科技,2018,25(4):463-466.
[10]黄丽,王春,李言,等.针刺预处理对脑缺血大鼠小脑浦肯野细胞动作电位的影响[J].南方医科大学学报,2018,38(6):677-682.
[11]张松兴,徐月英,邱泽玲.针刺预处理研究进展[J].辽宁中医药大学学报,2007,9(5):72-74.
[12]谢俊,陈泽斌,吴松,等.不同腧穴配伍电针防治大鼠心肌缺血损伤作用的比较[J].针刺研究,2017,42(2):131-135.
[13]徐文博,唐关敏,胡惠林,等.针刺预处理对缺血再灌注损伤大鼠心肌内质网应激的影响[J].中国中医药科技,2018,25(5):620-623.
[14]黄玉栋,聂焱,王志强,等.颅针对脑缺血再灌注损伤大鼠氧化应激的影响[J].上海针灸杂志,2016,35(12):1473-1476.
[15]徐芬,梁凤霞,陈瑞,等.标本配穴电针对胰岛素抵抗大鼠胰岛素敏感性及骨骼肌SIRT1蛋白表达的影响[J].中医杂志,2014,55(4):328-332.
[16]陈泽斌,邹峰,袁芳,等.针刺预处理对脑缺血再灌注大鼠顶皮质NR2A和NR2B蛋白表达的影响[J].中国中西医结合急救杂志,2005,12(2):79-83.
[17]王颖.针刺对脑缺血再灌注模型大鼠ROCK表达影响的动态观察[J].上海针灸杂志,2015,34(4):367-370.
[18]李珩,肖宁,邵明璐,等.针刺内关预处理干预心肌缺血再灌注大鼠实验效应的系统评价[J].中国中医基础医学杂志,2016,22(12):1665-1669.
[19]陈泽斌,柯晖,邹锋.针刺预处理抗脑缺血再灌注损伤作用及其分子机制[J].中医药学刊,2006,24(7):1246-1249.
[20]Paraíso AF,Mendes KL,Santos SH.Brain activation of SIRT1:role in neuropathology[J].Mol Neurobiol,2013,48(3):681-689.
[21]Samuel SM,Thirunavukkarasu M,Penumathsa SV,et al.Akt/FOXO3a/SIRT1-mediated cardioprotection by ntyrosol against ischemic stress in rat in vivo model of myocardial infarction:switching gears toward survival and longevity[J].J Agric Food Chem,2008,56(20):9692-9698.
[22]郭燕世.热量限制与白藜芦醇抗衰老的研究进展[J].生命科学进展,2011,42(3):161-164.
[23]Hernández-Jiménez M,Hurtado O,Cuartero MI,et al.Silent information regulator 1 protects the brain against cerebral ischemic damage[J].Stroke,2013,44(8):2333-2337.
[24]Miao Y,Zhao S,Gao Y,et al.Curcumin pretreatment attenuates inflammation and mitochondrial dysfunction in experimental stroke:the possible role of Sirt1signaling[J].Brain Res Bull,2016,121:9-15.
[25]Chen WK,Tsai YL,Shibu MA,et al.Exercise training augments Sirt1-signaling and attenuates cardiac inflamemation in D-galactose induced-aging rats[J].Aging(Albany NY),2018,10(12):4166-4174.
[26]Gupta AK,Choi S.Integrated Ligand and StructureBased Investigation of Structural Requirements for Silent Information Regulator 1(SIRT1)Activation[J].Curr Top Med Chem,2018,18(27):2313-2324.
[27]CantóC,Auwerx J.PGC-1alpha,SIRTI and AMPK,an energy sensing network that eonirols energy expenditure[J].Curr Opin Lipidol,2009,20(2):98-105.
[28]Wareski P,Vaarmann A,Choubey V,et al.PGC-1{alpha}and PGC-1{beta}regulate mitochondrial density in neurons[J].J Biol Chem,2009,284(32):21379-21385.