针刺“人中、内关”组穴对脑出血后脑红蛋白表达动态变化及线粒体形态结构变化影响实验研究
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摘要
目的:观察急性脑出血(Intracerebral hemorrhage,ICH)大鼠血肿周围脑组织脑红蛋白(Neuroglobin,NGB)随时间的动态变化规律、ICH后线粒体形态学变化以及针刺"人中、内关"组穴对上述观察指标的影响,探讨针刺"人中、内关"组穴治疗ICH的可能作用机制。方法:采用自体血注入法制作大鼠脑出血模型,选用70只健康雄性SD大鼠随机分为5组:空白对照组(n=5)、假手术组(n=5)、模型组(n=20)、穴位针刺组(n=20)、非穴针刺组(n=20)。造模后的每组再随机分为6 h、24 h、48 h、72 h 4个亚组。穴位针刺组针刺双侧"内关"和"人中"组穴。采用Bederson评分法对术后大鼠神经功能缺损程度进行评分符合标准纳入实验;电镜观察脑出血大鼠血肿周围组织线粒体超微结构;运用Western Blot法测定各时间点各组大鼠脑组织NGB相对表达量及其变化趋势。结果:(1)针刺"人中、内关"组穴能明显改善ICH后血肿周围组织线粒体损伤情况。(2)针刺"人中、内关"组穴能促进NGB高表达,与同时间点模型组相比,差异有统计学意义(P<0.05)。结论:根据本实验结论推断,针刺"人中、内关"组穴治疗ICH可能通过提高血肿周围组织NGB表达、改善血肿周围组织线粒体损伤程度、保护线粒体结构功能等途径,改善脑组织损伤和缺血缺氧状态,发挥保护神经细胞的作用。
Objective:To observe the acute cerebral hemorrhage rats brain tissue around hematoma NGB and the mitochondrial morphology dynamic changes with the time and the effects of acupuncture on Renzhong and Neiguan acupoints on the above indexes. To explore the possible mechanism of acupuncture in the treatment of cerebral hemorrhage. Methods:The model of ICH rats was established by autologous blood injection. 70 healthy male SD rats were randomly divided into five groups:control group,shamoperated group,model group,acupoint acupuncture group,non-acupoint acupuncture group,each group was randomly divided into four sub-groups:6 h,24 h,48 h and 72 h. Bederson scoring method was used to evaluate the degree of nerve function defect in postoperative rats;The ultrastructure of mitochondria around hematoma in ICH rats was observed by electron microscopy;Western Blot was used to determine the relative expression and variation trend of NGB in brain tissue of rats at different time points. Results:(1)Acupuncture on Renzhong and Neiguan acupoints can significantly improve the mitochondrial damage of surrounding tissue after ICH.(2)Acupuncture on Renzhong and Neiguan acupoints can promote high expression of NGB,and compared with the model group at the same time point,the difference was statistically significant(P<0.05). Conclusion:Based on this experiment,the results can be inferred that acupuncture on Renzhong and Neiguan acupoints to treat ICH may by increasing hematoma surrounding tissue expression of NGB,improve tissue around hematoma mitochondrial damage and protect the mitochondria structure function,improve the brain tissue damage and ischemia oxygen deficit,protect the nerve cells.
Objective:To observe the acute cerebral hemorrhage rats brain tissue around hematoma NGB and the mitochondrial morphology dynamic changes with the time and the effects of acupuncture on Renzhong and Neiguan acupoints on the above indexes. To explore the possible mechanism of acupuncture in the treatment of cerebral hemorrhage. Methods:The model of ICH rats was established by autologous blood injection. 70 healthy male SD rats were randomly divided into five groups:control group,shamoperated group,model group,acupoint acupuncture group,non-acupoint acupuncture group,each group was randomly divided into four sub-groups:6 h,24 h,48 h and 72 h. Bederson scoring method was used to evaluate the degree of nerve function defect in postoperative rats;The ultrastructure of mitochondria around hematoma in ICH rats was observed by electron microscopy;Western Blot was used to determine the relative expression and variation trend of NGB in brain tissue of rats at different time points. Results:(1)Acupuncture on Renzhong and Neiguan acupoints can significantly improve the mitochondrial damage of surrounding tissue after ICH.(2)Acupuncture on Renzhong and Neiguan acupoints can promote high expression of NGB,and compared with the model group at the same time point,the difference was statistically significant(P<0.05). Conclusion:Based on this experiment,the results can be inferred that acupuncture on Renzhong and Neiguan acupoints to treat ICH may by increasing hematoma surrounding tissue expression of NGB,improve tissue around hematoma mitochondrial damage and protect the mitochondria structure function,improve the brain tissue damage and ischemia oxygen deficit,protect the nerve cells.
引文
[1]Van Matre E T,Sherman D S,Kiser T H. Management of intracerebral hemorrhage-use of statins[J]. Vasc Health Risk Manag,2016,12:153-161.
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[6]韩佳炜,王淑华,李桂平,等.大鼠脑出血动物模型的制作技巧[J].中国实用神经疾病杂志,2018(7):707-711.
[7]Bederson J B,Pitts L H,Tsuji M,et al. Rat middle cerebral artery occlusion:evaluation of the model and development ofa neurologic examination[J]. Stroke,1986,17(3):472-476.
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[11]王晓光,樊小农,常女子,等.针刺水沟穴治疗脑梗死机制的研究进展[J].针灸临床杂志,2014(7):74-76.
[12]王琳,刘新桥,于涛.针刺人中、内关穴对心肺复苏后家兔脑线粒体呼吸功能的影响[J].天津中医药,2011(2):122-124.
[13]吕有魁,何宗宝.人中穴“醒脑开窍”作用机理的研究进展[J].中国中医基础医学杂志,2016(1):147-149.
[14]Burmester T,Weich B,Reinhardt S,et al. A vertebrate globin expressed in the brain[J]. Nature,2000,407(6803):520-523.
[15]颜虹,贺平,武姿含,等.针刺大椎、百会、人中穴对大鼠脑缺血再灌注损伤后不同时间点海马脑红蛋白表达的影响[J].湖南中医药大学学报,2018,38(2):165-168.
[16]LIU Y,LI B,LI Q,et al. Neuroglobin up-regulation after ischaemic pre-conditioning in a rat model of middle cerebral artery occlusion[J]. Brain Inj,2015,29(5):651-657.
[17]潘冬生,章翔,冯新莉,等.脑出血大鼠脑红蛋白表达与脑含水量变化[J].中华神经外科疾病研究杂志,2007(4):349-351.
[18]ZHANG L,LI L H,QU Y,et al. Neuroglobin and hypoxicischemic brain bamage[J]. Zhongguo Dang Dai Er Ke Za Zhi,2008,10(2):265-268.
[19]WANG X,LIU J,ZHU H,et al. Effects of neuroglobin overexpression on acute brain injury and long-term outcomes after focal cerebral ischemia[J]. Stroke,2008,39(6):1869-1874.
[20]张蓓,李亚军,张世俊,等.脑红蛋白在脑梗死大鼠脑组织中的表达及其神经保护作用机制[J].吉林大学学报:医学版,2012(6):1114-1118.
[21]LIU Z F,ZHANG X,QIAO Y X,et al. Neuroglobin protects cardiomyocytes against apoptosis and cardiac hypertrophy induced by isoproterenol in rats[J]. Int J Clin Exp Med,2015,8(4):5351-5360.
[22]Hundahl C A,Kelsen J,Hay-Schmidt A. Neuroglobin and cytoglobin expression in the human brain[J]. Brain Struct Funct,2013,218(2):603-609.
[23]夏翠萍.急性脑梗死患者监测Ngb、BDNF和MBP的临床意义[J].临床误诊误治,2016(12):81-84.
[24]游艳.脑出血中PGC-1α的作用和机制[D].重庆:重庆医科大学,2016.
[2]Adeoye O,Broderick J P. Advances in the management of intracerebral hemorrhage[J]. Nat Rev Neurol,2010,6(11):593-601.
[3]Feigin V L,Lawes C M,Bennett D A,et al. Worldwide stroke incidence and early case fatality reported in 56 population-based studies:a systematic review[J]. Lancet Neurol,2009,8(4):355-369.
[4]贾建平,陈生弟.神经病学[M].7版.北京:人民卫生出版社,2013:259-266.
[5]van Asch C J,Luitse M J,Rinkel G J,et al. Incidence,case fatality,and functional outcome of intracerebral haemorrhage over time,according to age,sex,and ethnic origin:a systematic review and meta-analysis[J]. Lancet Neurol,2010,9(2):167-176.
[6]韩佳炜,王淑华,李桂平,等.大鼠脑出血动物模型的制作技巧[J].中国实用神经疾病杂志,2018(7):707-711.
[7]Bederson J B,Pitts L H,Tsuji M,et al. Rat middle cerebral artery occlusion:evaluation of the model and development ofa neurologic examination[J]. Stroke,1986,17(3):472-476.
[8]李忠仁.实验针灸学[M].北京:中国中医药出版社,2003:425-431.
[9]华兴邦.大白鼠穴位图谱的研究[J].实验动物与动物实验,1991(1):13-15.
[10]杨兆辉,崔立金,孙亚新,等.利用功能性磁共振成像针刺人中穴位所致大脑皮层神经生理兴奋的现象[J].中国医药指南,2012(32):257-258.
[11]王晓光,樊小农,常女子,等.针刺水沟穴治疗脑梗死机制的研究进展[J].针灸临床杂志,2014(7):74-76.
[12]王琳,刘新桥,于涛.针刺人中、内关穴对心肺复苏后家兔脑线粒体呼吸功能的影响[J].天津中医药,2011(2):122-124.
[13]吕有魁,何宗宝.人中穴“醒脑开窍”作用机理的研究进展[J].中国中医基础医学杂志,2016(1):147-149.
[14]Burmester T,Weich B,Reinhardt S,et al. A vertebrate globin expressed in the brain[J]. Nature,2000,407(6803):520-523.
[15]颜虹,贺平,武姿含,等.针刺大椎、百会、人中穴对大鼠脑缺血再灌注损伤后不同时间点海马脑红蛋白表达的影响[J].湖南中医药大学学报,2018,38(2):165-168.
[16]LIU Y,LI B,LI Q,et al. Neuroglobin up-regulation after ischaemic pre-conditioning in a rat model of middle cerebral artery occlusion[J]. Brain Inj,2015,29(5):651-657.
[17]潘冬生,章翔,冯新莉,等.脑出血大鼠脑红蛋白表达与脑含水量变化[J].中华神经外科疾病研究杂志,2007(4):349-351.
[18]ZHANG L,LI L H,QU Y,et al. Neuroglobin and hypoxicischemic brain bamage[J]. Zhongguo Dang Dai Er Ke Za Zhi,2008,10(2):265-268.
[19]WANG X,LIU J,ZHU H,et al. Effects of neuroglobin overexpression on acute brain injury and long-term outcomes after focal cerebral ischemia[J]. Stroke,2008,39(6):1869-1874.
[20]张蓓,李亚军,张世俊,等.脑红蛋白在脑梗死大鼠脑组织中的表达及其神经保护作用机制[J].吉林大学学报:医学版,2012(6):1114-1118.
[21]LIU Z F,ZHANG X,QIAO Y X,et al. Neuroglobin protects cardiomyocytes against apoptosis and cardiac hypertrophy induced by isoproterenol in rats[J]. Int J Clin Exp Med,2015,8(4):5351-5360.
[22]Hundahl C A,Kelsen J,Hay-Schmidt A. Neuroglobin and cytoglobin expression in the human brain[J]. Brain Struct Funct,2013,218(2):603-609.
[23]夏翠萍.急性脑梗死患者监测Ngb、BDNF和MBP的临床意义[J].临床误诊误治,2016(12):81-84.
[24]游艳.脑出血中PGC-1α的作用和机制[D].重庆:重庆医科大学,2016.
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