电针百会、水沟穴上调脓毒性脑病大鼠脑红蛋白的表达
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摘要
目的观察并探讨大鼠出现脓毒性脑病时脑红蛋白(Neuroglobin,Ngb)的表达及电针百会、水沟穴对脓毒性脑病大鼠Ngb表达的影响。方法本研究采用经CLP造模成功的脓毒症大鼠通过观察大鼠神经行为学改变制作脓毒性脑病模型。将大鼠分为假手术组、脓毒性脑病组和脓毒性脑病+电针治疗组。取大鼠穴位百会、水沟穴分别沿皮刺入2 mm并接电极,另一电极夹在大鼠右耳尖端。疏密波,频率2 Hz/15 Hz,电流强度1 m A,以大鼠右耳微颤为电针刺激有效的标志,每12 h治疗30 min。2个组分别持续24 h和72 h。通过对大鼠脑组织切片及冰上取材,在24 h和72 h行免疫组化检测Ngb、Western blot检测Ngb蛋白表达水平及RealTime PCR检测Ngb mRNA基因表达水平,以测定电针百会、水沟穴对脓毒性脑病大鼠Ngb表达的影响。结果和假手术组比较,脓毒性脑病24 h组及72 h组的Ngb蛋白表达水平均明显上调(P<0.01);电针干预脓毒性脑病24 h组和非电针干预24 h组比较,Ngb蛋白表达明显上调(P<0.01);电针干预脓毒性脑病72 h组和非电针干预72 h组比较,Ngb蛋白表达稍有下调(P<0.01)。结论脓毒性脑病中Ngb表达上调,电针干预对脓毒性脑病有治疗作用,24 h最为明显,显示及时电针治疗脓毒性脑病效果明显,主要是通过上调Ngb表达起作用。
Objective To observe and investigate the expression of neuroglobin(Ngb) in rats with septic encephalopathy and the effect of electroacupuncture at Baihui and Shuigou acupoints on the expression of neuroglobin(Ngb) in rats with septic encephalopathy. Methods In this study, a rat model of sepsis encephalopathy was established by observing neurobehavioral changes in rats with sepsis successfully established by CLP. The rats were divided into sham operation group, septic brain disease group and septic brain disease + electroacupuncture treatment group. Electroacupuncture for Baihui, Shuigou points along the skin prick into 2 mm were performed, and the other electrode was caught in the right ear tip of the rats.Density wave frequency was 2 Hz/15 Hz, current intensity was 1 mA, and the rat's right ear microfibrillation were taken as a sign of effective electroacupuncture stimulation. Treatment was performed 30 minutes after per 12 hours. The two groups lasted 24 h and 72 h respectively. The brain tissue of rats was obtained, Ngb was detected by immunohistochemistry at 24 h and 72 h, and expression of Ngb protein level was detected by Western blot, Ngb mRNA gene was detected by RealTime PCR. Results Compared with the shamoperated group, the expression of Ngb protein was significantly up-regulated in the 24-hour septicencephalopathy group and the 72-hour group(P<0.01); the expression of Ngb protein was significantly increased in the 24-hour septic-encephalopathy group and the 24-hour non-electroacupuncture group(P<0.01); electro-acupuncture intervention 72 h septic brain disease group and non-electroacupuncture intervention 72 h group, Ngb protein expression was slightly down(P <0.01). Conclusion The expression of Ngb is up-regulated in septicencephalopathy, which means that electroacupuncture intervention has a therapeutic effect on septic brain disease, and it is most obvious at 24 hours. It shows that electroacupuncture is effective for the treatment of septic encephalopathy in time, mainly by up-regulating the expression of Ngb.
Objective To observe and investigate the expression of neuroglobin(Ngb) in rats with septic encephalopathy and the effect of electroacupuncture at Baihui and Shuigou acupoints on the expression of neuroglobin(Ngb) in rats with septic encephalopathy. Methods In this study, a rat model of sepsis encephalopathy was established by observing neurobehavioral changes in rats with sepsis successfully established by CLP. The rats were divided into sham operation group, septic brain disease group and septic brain disease + electroacupuncture treatment group. Electroacupuncture for Baihui, Shuigou points along the skin prick into 2 mm were performed, and the other electrode was caught in the right ear tip of the rats.Density wave frequency was 2 Hz/15 Hz, current intensity was 1 mA, and the rat's right ear microfibrillation were taken as a sign of effective electroacupuncture stimulation. Treatment was performed 30 minutes after per 12 hours. The two groups lasted 24 h and 72 h respectively. The brain tissue of rats was obtained, Ngb was detected by immunohistochemistry at 24 h and 72 h, and expression of Ngb protein level was detected by Western blot, Ngb mRNA gene was detected by RealTime PCR. Results Compared with the shamoperated group, the expression of Ngb protein was significantly up-regulated in the 24-hour septicencephalopathy group and the 72-hour group(P<0.01); the expression of Ngb protein was significantly increased in the 24-hour septic-encephalopathy group and the 24-hour non-electroacupuncture group(P<0.01); electro-acupuncture intervention 72 h septic brain disease group and non-electroacupuncture intervention 72 h group, Ngb protein expression was slightly down(P <0.01). Conclusion The expression of Ngb is up-regulated in septicencephalopathy, which means that electroacupuncture intervention has a therapeutic effect on septic brain disease, and it is most obvious at 24 hours. It shows that electroacupuncture is effective for the treatment of septic encephalopathy in time, mainly by up-regulating the expression of Ngb.
引文
[1]Papadopoulos MC,Davies DC,Moss RF,et al.Pathophysiology of septic encephalopathy:a review[J].Crit Care Med,2000,28(8):3019-3024.
[2]Moens L,Dewilde S.Globins in the brain[J].Nature,2000,407(6803):461-462.
[3]Burmester T,Weich B,Reinhardt S,et al.A vertebrate globin expressed in the brain[J].Nature,2000,407(6803):520-523.
[4]Hundahl CA,Allen GC,Nyengaard JR,et a1.Neuroglobin in the Rat Brain:localization[J].Neuroendbcfinology,2008,88(3):173-182.
[5]Shang A,Zhou D,Wang L,et al.Increased neuroglobin levels in the cerebral cortex and serum after ischemia-reperfusion insults[J].Brain Res,2006,1078(1):219-226.
[6]张丽娜.脓毒症相关性脑病的临床及基础研究[D].湖南:中南大学,2012:51-61.
[7]Wang X,Liu J,Zhu H,et al.Effects of neuroglobin overexpression on acute brain injury and brain-term outcomes after focal cerebral ischemia[J].Stroke,2008,39(6):1869-1874.
[8]李莉.脑红蛋白对神经元突起生长的作用及其潜在机制[D].湖北:华中科技大学,2013:44-47.
[9]张丽娜,艾宇航,郭曲练,等.脓毒症相关性脑病大鼠动物模型的建立[J].中华急诊医学杂志,2013,22(7):731-736.
[10]Kamata Y,Shiraga H,Tai A,et al.Induction of neurite outgrowth in PC12 cells by the medium-chain fatty acid octa[J].Neuroscience,2007,146(3):1073-1081.
[11]Fordel E,Thijs L,Moens L,et al.Neuroglobin and cytoglobin expression in mice.Evidence for a correlation with reactive oxygen species scavenging[J].FEBS J,2007,274(5):1312-1317.
[12]张镛,刘振芳,付庆喜,等.脑缺血和NBP预处理对脑缺血沙鼠NGB和mit-Na+,K+-ATP酶活性的影响[J].山东医药,2007,47(1):1-2.
[13]Garofalo T,Ferri A,Sorice M,et al.Neuroglobin overexpression plays a pivotal role in neuroprotection through mitochondrial raft-like microdomains in neuroblastoma SK-N-BE2 cells[J].Mol Cell Neurosci,2018,88(4):167-176.
[14]谢亚宁,王枫,王强,等.电针对脑缺血再灌注大鼠脑红蛋白的影响[J].针刺研究,2012,37(5):380-384.
[15]Yao WP,Wang S,Han L,et al.Effects of different frequencies of electroacupuncture at Shuigou(GV 26)on recovery of motor function in rats with focal cerebral ischemic injury[J].J Tradit Chin Med,2012,32(1):99-104.
[2]Moens L,Dewilde S.Globins in the brain[J].Nature,2000,407(6803):461-462.
[3]Burmester T,Weich B,Reinhardt S,et al.A vertebrate globin expressed in the brain[J].Nature,2000,407(6803):520-523.
[4]Hundahl CA,Allen GC,Nyengaard JR,et a1.Neuroglobin in the Rat Brain:localization[J].Neuroendbcfinology,2008,88(3):173-182.
[5]Shang A,Zhou D,Wang L,et al.Increased neuroglobin levels in the cerebral cortex and serum after ischemia-reperfusion insults[J].Brain Res,2006,1078(1):219-226.
[6]张丽娜.脓毒症相关性脑病的临床及基础研究[D].湖南:中南大学,2012:51-61.
[7]Wang X,Liu J,Zhu H,et al.Effects of neuroglobin overexpression on acute brain injury and brain-term outcomes after focal cerebral ischemia[J].Stroke,2008,39(6):1869-1874.
[8]李莉.脑红蛋白对神经元突起生长的作用及其潜在机制[D].湖北:华中科技大学,2013:44-47.
[9]张丽娜,艾宇航,郭曲练,等.脓毒症相关性脑病大鼠动物模型的建立[J].中华急诊医学杂志,2013,22(7):731-736.
[10]Kamata Y,Shiraga H,Tai A,et al.Induction of neurite outgrowth in PC12 cells by the medium-chain fatty acid octa[J].Neuroscience,2007,146(3):1073-1081.
[11]Fordel E,Thijs L,Moens L,et al.Neuroglobin and cytoglobin expression in mice.Evidence for a correlation with reactive oxygen species scavenging[J].FEBS J,2007,274(5):1312-1317.
[12]张镛,刘振芳,付庆喜,等.脑缺血和NBP预处理对脑缺血沙鼠NGB和mit-Na+,K+-ATP酶活性的影响[J].山东医药,2007,47(1):1-2.
[13]Garofalo T,Ferri A,Sorice M,et al.Neuroglobin overexpression plays a pivotal role in neuroprotection through mitochondrial raft-like microdomains in neuroblastoma SK-N-BE2 cells[J].Mol Cell Neurosci,2018,88(4):167-176.
[14]谢亚宁,王枫,王强,等.电针对脑缺血再灌注大鼠脑红蛋白的影响[J].针刺研究,2012,37(5):380-384.
[15]Yao WP,Wang S,Han L,et al.Effects of different frequencies of electroacupuncture at Shuigou(GV 26)on recovery of motor function in rats with focal cerebral ischemic injury[J].J Tradit Chin Med,2012,32(1):99-104.
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