从NOX4与炎性因子角度探讨清热、活血组分联用治疗缺血性脑卒中火毒证大鼠脑组织损伤的机制  

Investigation of the mechanism of combining the components of heat-clearing and blood-activating for the treatment of cerebral tissue lesion in rats with cerebral ischemic stroke of heat-toxin syndrome from the aspect of NOX4 and inflammatory factors

作　　者：王建伟[1] 郭蓉娟[2] 刘雪梅[3] 王凤丽[3] 闫妍[3] 郑宏[3] 梁晓[1] 张允岭[2] WANG Jian-wei GUO Rong-juan LIU Xue-mei WANG Feng-li YAN Yan ZHENG Hong LIANG Xiao ZHANG Yun-ling(Beijing University of Chinese Medicine, Beijing 100029, China Dongfang Hospital Beijing University of Chinese Medicine, Beijing 100078 Experimental Center, Dongfang Hospital Beijing University of Chinese Medicine, Beijing 100078)

机构地区：[1]北京中医药大学,北京100029 [2]北京中医药大学东方医院,北京100078 [3]北京中医药大学东方医院实验中心,北京100078

出　　处：《北京中医药》2017年第7期594-599,共6页Beijing Journal of Traditional Chinese Medicine

基　　金：国家国际科技合作专项项目(2015DFA31130);国家重点基础研究发展计划项目(973项目)(2012CB518406);国家自然科学基金项目(811173233)

摘　　要：目的从NOX4与炎性因子角度探讨清热、活血组分联用治疗缺血性脑卒中火毒证大鼠脑组织损伤的机制。方法采用腹腔注射角叉菜胶复合线栓致大脑中动脉闭塞复制大鼠缺血性脑卒中火毒证模型。将SD大鼠随机分为8组:空白对照组(N组)、假手术组(S组)、模型组(缺血1.5 h再灌注24 h,M组)、清热组分苦碟子注射液3.6 mL/kg组(A组)、活血组分血栓通注射液40 mg/kg组(B组)、苦碟子注射液1.8 mL/kg+血栓通注射液20 mg/kg组(C组)、苦碟子注射液1.8 mL/kg+血栓通注射液80 mg/kg组(D组)、苦碟子注射液7.2 mL/kg+血栓通注射液20 mg/kg组(E组)。Longa5级评分法进行大鼠神经功能评分;TTC染色法检测大鼠脑梗死面积百分比;免疫组化法检测梗死侧大脑皮层中IL-1β、TNF-α的阳性细胞表达;Western Blot法检测梗死侧大脑皮层中NOX4蛋白表达情况。结果神经功能评分示:S组未见神经功能缺损,M组神经功能缺损最重;与M组相比,A组、B组神经功能损伤减轻(P<0.05);与A组、B组相比,C组、E组神经功能缺损减轻,但差异无统计学意义(P>0.05),D组神经功能缺损减轻最为明显,差异有统计学意义(P<0.05)。TTC染色示:S组未见白色梗死灶,M组白色梗死区域明显;与M组相比,A组、B组白色梗死区域减少,梗死体积百分比减小(P<0.05,P<0.01);与A组、B组相比,C组、E组白色梗死区域减少,梗死体积百分比减小(P<0.05),D组脑梗死体积百分比最小(P<0.01)。免疫组化示:与S组相比,M组大脑皮层IL-1β及TNF-α阳性细胞数增多,黄色深染,凋亡细胞增多(P<0.01);与M组相比,A组、B组IL-1β及TNF-α阳性细胞数减少,颜色变浅;与A组、B组相比,C组、D组IL-1β阳性细胞表达量不同程度减少(P<0.01),E组无明显差异(P>0.05);与A组、B组相比,C组、D组、E组TNF-α阳性细胞表达量均不同程度减少(P<0.01)。Western Blot示:与S组相比,M组大鼠缺血侧大脑皮层中NOX4蛋白表达量显著增多(P<0.01);与MObjective To study the mechanism of combining the components for heat-clearing and blood-activating in the treatment of cerebral tissue lesion in rats with acute cerebral infarction of heat-toxin syndrome from the aspect of NOX4 and inflammatory factors.Methods Animal model of acute cerebral infarction of heat toxin pattern was established by using middle cerebral artery occlusion（MCAO） and peritoneal injection of carrageenan in rats.SD rats were randomly divided into eight groups：normal group（group N）,sham group（group S）,model group（ischemia for 1.5 h,reperfusion for 24 h,group M）,Kudiezi injection group of3.6 mL/kg（KDZ,for heat-clearing,group A）,Xueshuantong injection group of 40 mg/kg（XST,for blood activating,group B）,KDZ 1.8 mL/kg＋XST 20 mg/kg（group C）,KDZ 1.8 mL/kg ＋ XST 80 mg/kg（group D）,and KDZ 7.2 mL/kg＋XST 20 mg/kg（group E）.Longa 5 grade scoring method was used to measure the neurological function scores of rats;TTC staining method to measure the percentage of cerebral infarct area in rats;immunohistochemical assay to measure the expression of positive cells of IL-1β and TNF-α in the cortex on the infarct side;Western Blot method to detect NOX4 protein expression in the cerebral cortex on the infarct side.Results Neurological function scores：the neurological impairment was not seen in group S,but it was most serious in group M;compared with group M,the neurological impairment was alleviated in group A and B（P〈0.05）;compared with group A and B,the neurological impairment was more alleviated in group C and E,but the difference was not statistically significant（P〈0.05）,the nerve functional impairment was most serious in group D,the difference was statistically significant（P〈0.05）.TTC staining results：the white infarct area was not seen in Group S,but obviously in Group M;compared with group M,the cerebral infarction area was more decreased and the infarct volume percentage got smaller in group A and group B（P〈0.05,P〈0.01）;compared w

关 键 词：急性脑梗死 火毒证 NOX4 炎性因子 清热 活血 

分 类 号：R277.7[医药卫生—中医学]