黄连解毒汤有效部位对脑缺血半暗带区星形胶质细胞活化及Cx43表达的影响  

Effect of active fraction from Huanglian Jiedu Decoction on activation of astrocytes and expression of Cx43 in cerebral ischemic penumbra

作　　者：张建[1] 龙建飞[1] 邹海艳[1] 张秋霞[1] 赵晖[1] 王蕾[1] 张楠[1] 边宝林[2] 赵海誉[2] 

机构地区：[1]首都医科大学中医药学院,北京100069 [2]中国中医科学院中药研究所,北京100700

出　　处：《中草药》2014年第13期1876-1882,共7页Chinese Traditional and Herbal Drugs

基　　金：国家科技支撑计划课题:中药有效成分群功效关联性评价技术研究(2008BAI51B02);北京市属高等学校高层次人才引进与培养计划-长城学者项目资助(CIT&TCD20140329);北京市属高等学校人才强教深化计划"中青年骨干人材培养计划"项目(PXM2011014226)

摘　　要：目的观察黄连解毒汤水提物及其有效部位(总生物碱、总黄酮、总环烯醚萜)对局灶性脑缺血半暗带区星形胶质细胞活化及缝隙连接蛋白43(connexin 43,Cx43)表达的影响。方法线栓法建立大鼠永久性大脑中动脉栓塞模型,雄性SD大鼠随机分成假手术组、模型组、黄连解毒汤水提物组(800 mg/kg)、总生物碱组(44 mg/kg)、总黄酮组(50 mg/kg)、总环烯醚萜组(80 mg/kg),连续ig给药7 d,每天给药1次。HE染色观察脑组织病理学改变;胶质纤维酸性蛋白(GFAP)免疫荧光染色观察星形胶质细胞活化;GFAP/Cx43免疫荧光双标染色检测Cx43表达;实时RT-PCR检测Cx43基因表达。结果模型组大鼠缺血半暗带区神经元数目减少,GFAP、Cx43基因及蛋白表达上调(P<0.01)。黄连解毒汤水提物及其3个有效部位(总生物碱、总黄酮、总环烯醚萜)均可增加脑缺血大鼠缺血半暗带区神经元数目(P<0.05、0.01);降低缺血半暗带区GFAP及Cx43基因、蛋白表达(P<0.05、0.01)。结论黄连解毒汤水提物及其有效部位(总生物碱、总黄酮、总环烯醚萜)通过抑制星形胶质细胞过度活化,干预Cx43表达,保护缺血半暗带区神经细胞。Objective To observe the effect of water extract and active fractions from Huanglian Jiedu Decoction（HJD）（total alkaloids,total favonoids,and total iridoids） on the activation of astrocytes and expression of connexin 43（Cx43） in ischemic penumbra of rats after focal cerebral ischemia.Methods The rat model of middle cerebral artery ischemia was established using suture-occluded method.The male SD rats were randomly divided into Sham-operation,model,water extract of HJD（800 mg/kg）,total alkaloids（44 mg/kg）,total flavonoids（50 mg/kg）,and total iridoids（80 mg/kg） groups.They were ig administered for continuous 7 d,once daily.The histopathologic changes of brain in rats were observed by HE staining.The expression of GFAP and Cx43 was detected by immunofluorescence assay and RT-PCR was used to detect the gene expression of Cx43.Results In the model group,the number of neurons in ischemic penumbra of rats was significantly decreased and the expression of GFAP,Cx43,and Cx43 protein was obviously increased compared with Sham-operation group（P〈 0.01）.The three active fractions from HJD（total alkaloids,total favonoids,and total iridoids） could increase the number of neurons in the ischemic penumbra of rats and decrease the expression of GFAP,Cx43,and Cx43 protein obviously compared with the model group（P〈 0.05,0.01）.Conclusion The active fractions（total alkaloids,total favonoids,and total iridoids） could inhibit the over activation of astrocytes and the expression of Cx43,so as to protect the neurons in ischemic penumbra.

关 键 词：黄连解毒汤 有效部位 脑缺血 缺血半暗带区 星形胶质细胞 缝隙连接蛋白43 胶质纤维酸性蛋白 

分 类 号：R285.5[医药卫生—中药学]