眼针与体针对脑缺血再灌注损伤大鼠72 h脑组织AQP4表达的差异研究
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摘要
目的:观察眼针与体针对72 h脑缺血再灌注损伤模型(CIRI)大鼠脑组织水通道蛋白4(Aquaporin4,AQP4)表达差异的影响,探讨眼针治疗脑缺血再灌注损伤机制与体针的差异。方法:采用线栓法制备SD大鼠脑缺血再灌注损伤模型,将48只SD大鼠随机分为正常对照组、72 h假手术组、72 h模型组、72 h穴区组、72 h体针组、72 h穴区外组。脑缺血再灌注后72 h采用Zea Longa评分法进行大鼠神经功能评分,电镜下观察脑组织神经细胞形态学变化,Western blot方法测定脑组织AQP4蛋白,实时荧光定量聚合酶链反应(RQ-PCR)方法以及测定脑组织AQP4mRNA表达。结果:与72 h模型组比较,72 h穴区组大鼠神经功能缺损评分下降明显;穴区组大脑皮质神经元细胞电镜下体积增大,核大,胞质丰富,线粒体、粗面内质网等肿胀明显减轻。脑组织AQP4蛋白及mRNA表达明显下调(P <0. 05)。眼针与体针比较无统计学差异。结论:眼针和体针都能改善大鼠72 h脑缺血再灌注损伤,机制与下调脑组织AQP4表达有关,两者差异不明显。
Objective: To study the effect of the eye acupuncture therapy on cerebral AQP4 expression in rats with cerebral ischemia-reperfusion injury for 72 hours and the related mechanism. Methods: We made the rat model of cerebral ischemia-reperfusion by the suture method. SD rats were randomly grouped into normal control group,model group,sham operation group,eye acupuncture group,point area group,body acupuncture group and point area group. After reperfusion for 72 hours,the neural function score was adopted by Zea Longa scoring marks. The morphological changes of nerve cells were observed under electron microscope. The methods of Western-blotting and RT-PCR were used to detect the changes of the AQP4 protein and AQP4 mRNA in rat brains after the eye acupuncture therapy. Results: Compared with the model group,the neurologic impairment score of the eye-acupuncture group got lower. The expressions of the AQP4 protein and AQP4 mRNA in rat brains after the eye acupuncture therapy were significantly reduced( P < 0. 05). However,there was no statistical difference between the eye acupuncture group and the body acupuncture group. Conclusion: Both the methods of eye acupuncture therapy and body acupuncture therapy can reduce the cerebral ischemia-reperfusion injury by inhibiting the expression of the cerebral AQP4 protein.
Objective: To study the effect of the eye acupuncture therapy on cerebral AQP4 expression in rats with cerebral ischemia-reperfusion injury for 72 hours and the related mechanism. Methods: We made the rat model of cerebral ischemia-reperfusion by the suture method. SD rats were randomly grouped into normal control group,model group,sham operation group,eye acupuncture group,point area group,body acupuncture group and point area group. After reperfusion for 72 hours,the neural function score was adopted by Zea Longa scoring marks. The morphological changes of nerve cells were observed under electron microscope. The methods of Western-blotting and RT-PCR were used to detect the changes of the AQP4 protein and AQP4 mRNA in rat brains after the eye acupuncture therapy. Results: Compared with the model group,the neurologic impairment score of the eye-acupuncture group got lower. The expressions of the AQP4 protein and AQP4 mRNA in rat brains after the eye acupuncture therapy were significantly reduced( P < 0. 05). However,there was no statistical difference between the eye acupuncture group and the body acupuncture group. Conclusion: Both the methods of eye acupuncture therapy and body acupuncture therapy can reduce the cerebral ischemia-reperfusion injury by inhibiting the expression of the cerebral AQP4 protein.
引文
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[7]张威,马贤德,田维柱.眼针“八区八穴”与“八区十三穴”眼针疗法对中风病患者临床疗效比较[J].辽宁中医药大学学报,2016,18(4):76-79.
[8]马贤德,孙宏伟.线栓法制备大鼠脑缺血再灌注模型的方法研究[J].中华中医药学刊,2009,27(6):1200-1201.
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[11]Zhu MX,Xing D,Lu ZH,et al.DDR1 may play a key role in destruction of the blood-brain barrier after cerebral ischemia-reperfusion[J].Neurosci Res,2015,96:14-19.
[12]贺伟,展宏刚.SIRT1对早期脑缺血大鼠水通道蛋白4表达的影响[J].中国病理生理杂志,2017,33(3):455-461
[13]牛彩虹,齐进冲,修宝新,等.大鼠脑缺血-再灌注损伤早期DWI参数和AQP4蛋白表达相关性研究[J].脑与神经疾病杂志,2016,24(10):617-623.
[14]Taya K,Marmarou CR,Okuno K,et al.Effect of secondary insultsupon aquaporin-4 water channels following experimental corticalcontusion in rats[J].J Neurotrauma,2010,27(1):229-239.
[15]江金文,刘会云,李美华.AQP4与创伤性脑水肿的研究进展[J].国际神经病学神经外科学杂志,2017,44(2):213-217.
[16]Miao R,Li CY.The research progress of AQP-dependent cell migration[J].Chin Pharmacol Bull,2011,27(5):601-605.
[17]许淑红,康辰,陈美灵,等.大鼠脑缺血/再灌注后早期AQP4的动态表达及其与脑水肿关系的研究[J].中国药理学通报,2016,32(10):1433-1441.
[18]王秀辉,郑咏秋.大鼠脑缺血再灌注后AQP4的表达与脑水肿的关系[J].中国实验方剂学杂志,2013,19(15):182-185.
[19]Hai-Ming Chen,Huan-Sen Huang.Ulinastatin attenuates cerebral ischemia-reperfusion injury in rats[J].Int J ClinExp Med,2014,7(5):1483-1489.
[20]陆立和,黄李平.中药怀牛膝及黄芪对重型颅脑损伤大鼠脑水肿及脑组织AQP4表达的影响[J].时珍国医国药,2014,25(10):2359-2361.
[2]Crane JM,Rossi A,Gupta T.Orthogonal array formation by human aquaporin-4:Examination of neuromyelitisoptica-associated aquaporin-4 poly morphisms[J].Journal of Neuroimmunology,2011,236:93-98.
[3]Chu H,Tang Y,Dong Q.Protection of vascular endothelial growth factor to brain edema followongintracerebral hemorrhage and its involved mechanisms:effect of aquaporin-4[J].PLos One,2013,8(6):65-65.
[4]Badaut J,Fukuda AM,Jullienne AJ,et al.Aquaporin and brain diseases[J].Biochim Biophys Acta,2014,1840(5):1554-1565.
[5]刘露阳,王鹏琴.眼针治疗中风偏瘫疗效meta分析[J].辽宁中医药大学学报,2017,19(2):102-104.
[6]王鹏琴,鞠庆波,宋哲,等.眼针带针康复法促进中风偏瘫患者运动功能恢复的临床研究[J].中国中医基础医学杂志,2016,22(4):534-536,560.
[7]张威,马贤德,田维柱.眼针“八区八穴”与“八区十三穴”眼针疗法对中风病患者临床疗效比较[J].辽宁中医药大学学报,2016,18(4):76-79.
[8]马贤德,孙宏伟.线栓法制备大鼠脑缺血再灌注模型的方法研究[J].中华中医药学刊,2009,27(6):1200-1201.
[9]Longa EZ,Weinstein PR,Carlson S,et al.Reversible middle cerebral artery occlusion without craniectomy in rats[J].Stroke,1989,20(1):84-91.
[10]彭静山.眼针疗法[M].沈阳:辽宁科学技术出版社,1990:11.
[11]Zhu MX,Xing D,Lu ZH,et al.DDR1 may play a key role in destruction of the blood-brain barrier after cerebral ischemia-reperfusion[J].Neurosci Res,2015,96:14-19.
[12]贺伟,展宏刚.SIRT1对早期脑缺血大鼠水通道蛋白4表达的影响[J].中国病理生理杂志,2017,33(3):455-461
[13]牛彩虹,齐进冲,修宝新,等.大鼠脑缺血-再灌注损伤早期DWI参数和AQP4蛋白表达相关性研究[J].脑与神经疾病杂志,2016,24(10):617-623.
[14]Taya K,Marmarou CR,Okuno K,et al.Effect of secondary insultsupon aquaporin-4 water channels following experimental corticalcontusion in rats[J].J Neurotrauma,2010,27(1):229-239.
[15]江金文,刘会云,李美华.AQP4与创伤性脑水肿的研究进展[J].国际神经病学神经外科学杂志,2017,44(2):213-217.
[16]Miao R,Li CY.The research progress of AQP-dependent cell migration[J].Chin Pharmacol Bull,2011,27(5):601-605.
[17]许淑红,康辰,陈美灵,等.大鼠脑缺血/再灌注后早期AQP4的动态表达及其与脑水肿关系的研究[J].中国药理学通报,2016,32(10):1433-1441.
[18]王秀辉,郑咏秋.大鼠脑缺血再灌注后AQP4的表达与脑水肿的关系[J].中国实验方剂学杂志,2013,19(15):182-185.
[19]Hai-Ming Chen,Huan-Sen Huang.Ulinastatin attenuates cerebral ischemia-reperfusion injury in rats[J].Int J ClinExp Med,2014,7(5):1483-1489.
[20]陆立和,黄李平.中药怀牛膝及黄芪对重型颅脑损伤大鼠脑水肿及脑组织AQP4表达的影响[J].时珍国医国药,2014,25(10):2359-2361.