针刺对脑缺血大鼠血管生成和细胞外信号调节激酶1/2信号通路的影响
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摘要
目的:观察针刺对脑缺血大鼠血管生成和细胞外信号调节激酶1/2信号通路的影响。方法:将30只健康SD大鼠分为正常对照组、模型对照组、针刺组各10只。模型对照组以及针刺组给予Bannister's颈动脉引流法以建立大鼠脑缺血的动物模型。针刺组造模7 d后,在大鼠保持清醒的状况下,固定于自制鼠架上,予以百会穴平刺2 mm,捻转1 min增强针感,留针20 min;水沟穴点刺1 mm,捻转1 min增强针感,不留针;每日1次,每周6次,2周为1个疗程,共治疗1个疗程。其余两组不予以任何干预措施,至实验结束。比较各组大鼠情况、神经功能缺损量表评分、海马脑细胞凋亡百分率、椎体细胞数、血管内生长因子表达水平、血管密度、磷酸化氨基末端蛋白激酶(p-Jun N-ternuial kinase,p-JNK)以及磷酸化细胞外信号调节激酶(phosphorylated extacellular signal-regulated kinase,p-ERK1/2)蛋白表达比较。结果:治疗后,与模型对照组比较,针刺组治疗后7 d、治疗后14 d的神经功能缺损量表分数较低;与治疗后7 d比较,治疗后14 d针刺组神经功能缺损量表评分较低(P<0.05)。治疗后,与正常对照组比较,模型对照组和针刺组海马神经细胞凋亡率较高,锥体细胞存活数较低(P<0.05);与模型对照组比较,针刺组海马神经细胞凋亡率较低,锥体细胞存活数较高(P<0.05)。治疗后,与正常对照组比较,模型对照组以及针刺组血管内皮生长因子(vascular endothelial growth factor,VEGF)蛋白表达以及血管密度较低(P<0.05);与模型对照组比较,针刺组VEGF蛋白表达以及血管密度较高(P<0.05)。治疗后,与正常对照组比较,模型对照组以及针刺组p-JNK、p-ERK1/2蛋白表达较高(P<0.05);与模型对照组比较,针刺组p-JNK蛋白表达较低,p-ERK1/2蛋白表达较高(P<0.05)。结论:针刺能够上调脑缺血大鼠VEGF表达水平,下调海马神经细胞凋亡率,提高促进组织脑血管形成,调节p-JNK以及p-ERK1/2表达水平,从而减轻脑损害程度,显著促进脑缺血后神经功能修复,这可能是针刺治疗脑缺血损伤的机制之一。
objective: To explore the effects of acupuncture on angiogenesis and extracellular signal regulated kinase 1/2 signaling pathway in rats with focal cerebral ischemia. Method: 30 healthy SD rats were divided into normal control group,model control group and acupuncture group,with 10 rats in each group. The model group and the acupuncture group were given Bannister's carotid artery drainage to establish the animal model of cerebral ischemia in rats. After 7 days,keep rats awake of acupuncture group and fixed on the rat frame,to horizontal needling Baihui for 2 mm,twirling needle 1 min and retaining for 20 min; Rapid needling Shuigou for 1 min,twirling needle 1 min without retaining; Treat for once a day,6 times a week of 2 weeks. The other two groups did not receive any intervention until the end of the experiment. Neurological deficit scale,percentage of apoptotic cells in the hippocampus,number of vertebral cells,expression of intravascular growth factor,vascular density,p-JNK and p-ERK1/2 protein expression were compared. Results: After treatment,compared with the model control group,the NSS score of acupuncture group after 7 days treatment was lower. Compared with 7 days after treatment,the NSS score of acupuncture group was lower for 14 days after treatment( P < 0. 05). After treatment,compared with the normal control group,the apoptosis rate of hippocampal neurons in model group and acupuncture group was higher,and the number of pyramidal cells was lower( P < 0. 05); Compared with the model group,the apoptosis rate of hippocampal neurons in acupuncture group was lower,and the number of pyramidal cells was higher( P < 0. 05). After treatment,compared with that of the normal control group,the expression of VEGF protein in the model group and the acupuncture group was lower than that of the control group( P < 0. 05); Compared with the model group,the expression of VEGF protein and blood vessel density in the acupuncture group were higher than those in the control group( P < 0. 05). After treatment,compared with the normal control group,the expression of p-JNK and p-ERK1/2 protein in the model group and the acupuncture group was higher( P < 0. 05); Compared with the model group,the expression of p-JNK protein was lower in the acupuncture group,and the expression of p-ERK1/2 protein was higher( P < 0. 05). Conclusion: Acupuncture can increase expression of VEGF in cerebral ischemia rats,reduce hippocampal nerve cell apoptosis rate,improve and promote the formation of the cerebral vascular regulation of p-JNK and the expression level of p-ERK1/2,thereby reducing brain damage,significantly promoting the repair of nerve function after cerebral ischemia,which may be one of the mechanism of acupuncture on cerebral ischemic injury.
objective: To explore the effects of acupuncture on angiogenesis and extracellular signal regulated kinase 1/2 signaling pathway in rats with focal cerebral ischemia. Method: 30 healthy SD rats were divided into normal control group,model control group and acupuncture group,with 10 rats in each group. The model group and the acupuncture group were given Bannister's carotid artery drainage to establish the animal model of cerebral ischemia in rats. After 7 days,keep rats awake of acupuncture group and fixed on the rat frame,to horizontal needling Baihui for 2 mm,twirling needle 1 min and retaining for 20 min; Rapid needling Shuigou for 1 min,twirling needle 1 min without retaining; Treat for once a day,6 times a week of 2 weeks. The other two groups did not receive any intervention until the end of the experiment. Neurological deficit scale,percentage of apoptotic cells in the hippocampus,number of vertebral cells,expression of intravascular growth factor,vascular density,p-JNK and p-ERK1/2 protein expression were compared. Results: After treatment,compared with the model control group,the NSS score of acupuncture group after 7 days treatment was lower. Compared with 7 days after treatment,the NSS score of acupuncture group was lower for 14 days after treatment( P < 0. 05). After treatment,compared with the normal control group,the apoptosis rate of hippocampal neurons in model group and acupuncture group was higher,and the number of pyramidal cells was lower( P < 0. 05); Compared with the model group,the apoptosis rate of hippocampal neurons in acupuncture group was lower,and the number of pyramidal cells was higher( P < 0. 05). After treatment,compared with that of the normal control group,the expression of VEGF protein in the model group and the acupuncture group was lower than that of the control group( P < 0. 05); Compared with the model group,the expression of VEGF protein and blood vessel density in the acupuncture group were higher than those in the control group( P < 0. 05). After treatment,compared with the normal control group,the expression of p-JNK and p-ERK1/2 protein in the model group and the acupuncture group was higher( P < 0. 05); Compared with the model group,the expression of p-JNK protein was lower in the acupuncture group,and the expression of p-ERK1/2 protein was higher( P < 0. 05). Conclusion: Acupuncture can increase expression of VEGF in cerebral ischemia rats,reduce hippocampal nerve cell apoptosis rate,improve and promote the formation of the cerebral vascular regulation of p-JNK and the expression level of p-ERK1/2,thereby reducing brain damage,significantly promoting the repair of nerve function after cerebral ischemia,which may be one of the mechanism of acupuncture on cerebral ischemic injury.
引文
[1]袁磊.急性脑梗死患者血小板凝血因子Ⅷ升高与脑血栓形成的相关性研究[J].中国临床神经科学,2015,23(5):550-552.
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[7]赵海军,王媛,卢岩,等.针刺对缺血脑组织谷氨酸含量及其代谢型受体5 mRNA表达水平影响的实验研究[J].江苏中医药,2014,46(10):70-72.
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[9]张红星,杨敏,周利,等.头针对急性脑缺血再灌注大鼠的保护作用[J].神经损伤与功能重建,2008,3(6):377-380.
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[12]王睿,焦富英.针刺治疗脑卒中后吞咽障碍简况[J].实用中医内科杂志,2016,v.30(4):117-118.
[13]张海涛,张平,蒋传路,等.血管内皮生长因子在脑缺血再灌注损伤中的作用研究进展[J].疑难病杂志,2015,14(7):756-758.
[14]王卫东,王琳,邓丽影,等.内源性VEGF表达对缺血性脑损伤后成年大鼠海马齿状回神经发生影响的实验研究[J].中风与神经疾病,2009,26(4):388-391.
[15]黄宁芳.血管内皮生长因子对缺血性脑血管病的临床治疗研究进展[J].吉林医学,2015,36(8):1639-1640.
[16]刘俊娥,张继平.补阳还五汤抗脑缺血神经细胞凋亡实验研究进展[J].中国中医药信息杂志,2011,18(9):110-112.
[17]潘蓓,刘瑞珍,谢宝明.大鼠局灶性脑缺血再灌注后P-erk表达及神经元凋亡的变化[J].山西医科大学学报,2009,40(5):433-436.
[18]王婷婷,翟丽,郭云良.胡黄连苷Ⅱ对脑缺血损伤后细胞外信号调节蛋白激酶通路的影响及神经保护作用[J].中华行为医学与脑科学杂志,2016,25(2):97-102.
[19]曹畅,刘婷婷,蔡哲平,等.ERK1/2介导姜黄素抑制STS诱导神经元毒性损伤的作用[J].中国生化药物杂志,2015,35(4):1-4.
[20]邢变枝,陈晖,张苏明.缺血后处理对脑缺血再灌注损伤后ERK1/2和Akt磷酸化及神经细胞凋亡的影响[J].神经损伤与功能重建,2012,7(3):175-179.
[2]龚涛.氧化应激与缺血性卒中急性事件[J].中国卒中杂志,2010,5(2):134-138.
[3]肖慧,谷文萍,胡珏,等.Ephrin-B2促进大鼠局灶脑缺血再灌注后VEGF表达及血管新生[J].现代生物医学进展,2014,14(30):5824-5828.
[4]严洪新,徐恩.细胞外信号调节激酶与缺血性脑卒中[J].现代医学,2010,38(4):427-430.
[5]刘涓,谢瑞娟,石翠霞,等.现代康复训练结合中医针刺治疗小儿脑瘫临床疗效及对肢体运动功能与脑血流影响[J].辽宁中医药大学学报,2016,18(8):251-253.
[6]宋光,何蕾,王彬,等.黄芪激活ERK1/2信号转导通路抗心肌细胞再灌注损伤作用[J].中国体外循环杂志,2008,6(1):52-54.
[7]赵海军,王媛,卢岩,等.针刺对缺血脑组织谷氨酸含量及其代谢型受体5 mRNA表达水平影响的实验研究[J].江苏中医药,2014,46(10):70-72.
[8]陈晔,严晓丹,林斌强,等.电针对局部脑缺血大鼠脑内P物质表达的影响研究[J].福建中医药大学学报,2012,22(6):16-18.
[9]张红星,杨敏,周利,等.头针对急性脑缺血再灌注大鼠的保护作用[J].神经损伤与功能重建,2008,3(6):377-380.
[10]Weidner N.Current pathologic methods for measuring intratumoral microvessel density within breast carcinoma and other solid tumors[J].Breast Cancer Res Treat,1995,36(2):169-180.
[11]焦顺发,焦青峰.督脉是脊髓浅识[J].中国针灸,2006,26(10):761-761.
[12]王睿,焦富英.针刺治疗脑卒中后吞咽障碍简况[J].实用中医内科杂志,2016,v.30(4):117-118.
[13]张海涛,张平,蒋传路,等.血管内皮生长因子在脑缺血再灌注损伤中的作用研究进展[J].疑难病杂志,2015,14(7):756-758.
[14]王卫东,王琳,邓丽影,等.内源性VEGF表达对缺血性脑损伤后成年大鼠海马齿状回神经发生影响的实验研究[J].中风与神经疾病,2009,26(4):388-391.
[15]黄宁芳.血管内皮生长因子对缺血性脑血管病的临床治疗研究进展[J].吉林医学,2015,36(8):1639-1640.
[16]刘俊娥,张继平.补阳还五汤抗脑缺血神经细胞凋亡实验研究进展[J].中国中医药信息杂志,2011,18(9):110-112.
[17]潘蓓,刘瑞珍,谢宝明.大鼠局灶性脑缺血再灌注后P-erk表达及神经元凋亡的变化[J].山西医科大学学报,2009,40(5):433-436.
[18]王婷婷,翟丽,郭云良.胡黄连苷Ⅱ对脑缺血损伤后细胞外信号调节蛋白激酶通路的影响及神经保护作用[J].中华行为医学与脑科学杂志,2016,25(2):97-102.
[19]曹畅,刘婷婷,蔡哲平,等.ERK1/2介导姜黄素抑制STS诱导神经元毒性损伤的作用[J].中国生化药物杂志,2015,35(4):1-4.
[20]邢变枝,陈晖,张苏明.缺血后处理对脑缺血再灌注损伤后ERK1/2和Akt磷酸化及神经细胞凋亡的影响[J].神经损伤与功能重建,2012,7(3):175-179.