康脑液对脑缺血再灌注损伤大鼠血管内皮生长因子、脑源性神经生长因子、基质金属蛋白酶表达的影响

神经药理学报 ›› 2014, Vol. 4 ›› Issue (2): 16-21.

康脑液对脑缺血再灌注损伤大鼠血管内皮生长因子、脑源性神经生长因子、基质金属蛋白酶表达的影响

马建鹏, 常青, 薛茜, 邹玉安

1.河北北方学院，张家口，075000，中国
2.河北北方学院附属第一医院神经内科，张家口，075000，中国

出版日期:2014-04-26 发布日期:2014-06-27
通讯作者: 薛茜,女,教授,主任医师,硕士,硕士生导师;研究方向:缺血脑血管病;E-mail:xueqian6166@163.com
作者简介:马建鹏,女,硕士研究生在读；研究方向：缺血性脑血管病；E-mail：majianpeng45@163.com
基金资助:
河北省卫生厅课题基金资助项目( 20090591)

Effect of Kangnaoye on Expression of VEGF, BDNF, MMP-9 after Cerebral Ischemia Reperfusion in Rats

MA Jian-peng, CHANG-Qing, XUE-Qian, ZOU Yu-an

1 Hebei North University, Zhangjiakou, 075000, China
2 Department of Neurology, the First Affiliated Hospital of Hebei North University, Zhangjiakou, 075000, China

Online:2014-04-26 Published:2014-06-27
Contact: 薛茜,女,教授,主任医师,硕士,硕士生导师;研究方向:缺血脑血管病;E-mail:xueqian6166@163.com
About author:马建鹏,女,硕士研究生在读；研究方向：缺血性脑血管病；E-mail：majianpeng45@163.com
Supported by:
河北省卫生厅课题基金资助项目( 20090591)

摘要/Abstract

摘要： 目的：通过康脑液干预观察其对脑缺血再灌注损伤大鼠血管内皮生长因子(vascular endothelial growth factor ，VEGF) 、脑源性神经生长因子（brain derived neurotrophic factor ，BDNF）、基质金属蛋白酶（matrix metalloproteinase-9，MMP-9）表达的影响，探讨康脑液对脑缺血再灌注损伤的保护机制。方法：将雄性SD 大鼠随机分为假手术组、脑缺血再灌注模型组及康脑液28.6，14.3，7.15 g·kg-1·d-1剂量组（灌胃给药7 d），改进Longa等线栓法制备大鼠右侧大脑中动脉阻塞（middle cerebral artery occlusion，MCAO）再灌注模型。于缺血2 h后再灌注（分别于再灌注后6 h，12 h，24 h，72 h，7 d处死大鼠）；采用TTC染色法观察大鼠的脑梗死面积；免疫组织化学法观察大鼠脑组织VEGF、BDNF、MMP-9的表达。结果:比较各组缺血再灌注24 h大鼠脑梗死灶面积，发现28.6，14.3 g·kg-1·d-1剂量组较脑缺血再灌注模型组明显减小（P＜0.05）；与脑缺血再灌注模型组组相比，28.6，14.3 g·kg-1·d-1剂量组各时间点的VEGF 、BDNF表达量明显上调(P＜0.01)，MMP-9表达的阳性细胞明显减少（P ＜0.01），具有统计学意义。结论：康脑液可促进大鼠局灶性脑缺血后脑组织中VEGF，BDNF的表达，同时抑制脑内MMP-9的表达，缩小脑梗死面积，发挥对神经血管单元（Neurovascular Unit，NVU）的保护作用,减轻脑缺血再灌注损伤。

关键词: 康脑液, 神经血管单元, 血管内皮生长因子, 脑源性神经生长因子, 基质金属蛋白酶

Abstract: Objective: To observe the effect of Kangnaoye pretreatment on the expression of vascular endothelial growth factor (VEGF), brain derived neurotrophic factor (BDNF),matrix metalloproteinase-9 (MMP-9) in rats with cerebral ischemia-reperfusion injury, and further to explore the neurovacular unit protective mechanism of Kangnaoye against the injury of cerebral ischemia-reperfusion. Methods: Male Spraque-Dawley rats were randomly divided into: Sham operation group，ischemia-reperfusion group，Kangnaoye 28.6，14.3，7.15 g·kg -1·d-1 dose group (intragastric administration, 7 d). Improved Zea Longa method was used to prepare cerebral ischemia-reperfusion model of rats. Reperfusion after ischemia for 2 h（rats were sacrificed respectively after reperfusion 6 h，12 h，24 h，72 h，7 d）. TTC staining method was used for observing the infarct area of rat brain; Immunohistochemical method was used for observing the expression of VEGF, BDNF, MMP-9 in rats brain tissue. Results: The comparison of the infarct area between the groups 24h after reperfusion ischemic found that the Kny High, Middle dose group significantly decreased the infarct area (P<0.05), along with the unregulated expression of VEGF,BDNF（P<0.01）. The expression of MMP-9 was significantly decreased as compared to model group（P<0.01）. Conclusion: Kangnaoye promoted the expression of VEGF, BDNF in brain tissue, inhibited the expression of MMP-9, decreased the infarct area and protected neurovascular unit against cerebral ischemia reperfusion injury.

Key words: kangnaoye, neurovascular unit, vascular endothelial growth factor, brain derived neurotrophic factor, matrix metalloproteinase-9

中图分类号:

R965

马建鹏, 常青, 薛茜, 邹玉安. 康脑液对脑缺血再灌注损伤大鼠血管内皮生长因子、脑源性神经生长因子、基质金属蛋白酶表达的影响[J]. 神经药理学报, 2014, 4(2): 16-21.

MA Jian-peng, CHANG-Qing, XUE-Qian, ZOU Yu-an. Effect of Kangnaoye on Expression of VEGF, BDNF, MMP-9 after Cerebral Ischemia Reperfusion in Rats[J]. ACTA NEUROPHARMACOLOGICA, 2014, 4(2): 16-21.

参考文献

[1] Sun Xiu-lan, Hu Gang. ATP-sensitive potassium channels: a promising target for protecting neurovascular unit function in stroke[J].Clin Exp Pharmacol physiol, 2010, 37(2): 243-252.

[2] Gregory J del Zoppo. Stroke and neurovascular protection[J].N Engl J Med, 2006, 354(6): 553-555.

[3] 周赛男,蔺晓源,易健,等.补阳还五汤对脑缺血大鼠神经功能及细胞形态的影响[J].中国实验方剂学杂志, 2013, 19(2): 251-254.

[4] 石会娟,梁起保,薛茜,等.康脑液对脑缺血再灌注大鼠血清及脑组织中氨基酸含量的影响[J].天津医药, 2011, 39(10): 939-941.

[5] 赵志敏,肖志娟,薛茜,等.康脑液对脑缺血再灌注动物血管新生的影响[J].中国临床药理学, 2014, 30(10): 429-431.

[6] Wang Yong-quan, Guo Xin, Qiu Mei-hong, et al. VEGF overexpression enhances striatal neurogenesis in brain of adult rat after a transient middle cerebral artery occlusion[J].J Neurosci Res, 2007, 85(1): 73-82.

[7] Jin Kun-lin, Zhu Yong-hua, Sun Yun-juan, et al. Vascular endothelial growth factor (VEGF) stimulates neurogenesis in vitro and in vivo[J].Proc Natl Acad Sci USA, 2002, 99(18): 11946-11950.

[8] Feng S, Cen J, Huang Y, et al. Matrix metalloproteinase-2 and -9 secreted by leukemic cells increase the permeability of blood-brain barrier by disrupting tight junction proteins [J]. PLoS One, 2011, 6(8): e20599.

[9] Enrique Zea Longa, Philip R Weinstein, Sara Carlson, et al. Reversible middle cerebral artery occlusion without cranietomy in rats[J].Stroke, 1989, 20(1):84-91.

[10] Joshua B Bederson, Lawrence H Pitts, Miles Tsuji, et al. Rat middle cerebral artery occlusion: evaluation of the model and development of a neurologic examination [J].Stroke, 1986, 17(3): 472-476．

[11] Otilia M?rg?ritescu, Pirici D, M?rg?ritescu C L.VEGF expression in human brain tissue after acute ischemic stroke[J].Rom J Morphol Embryol, 2011, 52(4): 1283-1292.

[12] 赛音巴雅尔,林贵军,饶芝国,等.外源性血管内皮生长因子对大鼠脑血肿周围血管新生及神经功能恢复的影响[J].中华神经外科杂志, 2012, 28(5): 522-525.

[13] Guo Shu-zhen, Woo Jean Kim, Josephine Lok, et al. Neuropretection via matrix-troohic coupling between cerebral cells and neurons[J].Proc Natl Acad Sci USA, 2008, 105(21): 7582-7587.

[14] Hyun Kim, Li Qi, Barbara L Hempstead, et al. Paracrine and autocrine functions of brain- derived neurotrophic factor (BDNF)and nerve growth factor (NGF) in brain-der ivedendothe lial cells [J]. J Bio l Chem, 2004, 279(32): 33538-33546.

[15] 王强,张一,王榕,等.丹参多酚酸盐对缺血再灌注损伤大鼠脑组织BDNF和GDNF的影响[J].卒中与神经疾病, 2013, 5(20): 272-274.

[16] Eng H Lo, Turgay Dalkara, Michael A Moskowitz. Mechanisms, challenges and opportunities in stroke [J]. Nat Rev Neurosci, 2003, 4(5): 399-415.

[17] Li Hong, Wei Ming, Li Sheng-hui, et al. Increased CD147 and MMP-9 expression in the normal rat brain after gamma irradiation[J]. J Radiat Res, 2013, 54(1):27-35.