摘要
目的:观察黄连解毒汤水提物及3个有效部位(总生物碱、总黄酮、总环烯醚萜)对中动脉栓塞(MCAO)大鼠丘脑的保护作用。方法:线栓法建立MCAO大鼠模型,雄性SD大鼠随机分成假手术组、模型组、黄连解毒汤水提物(800 mg·kg-1)、总生物碱组(44 mg·kg-1)、总黄酮组(50 mg·kg-1)、总环烯醚萜组(80 mg·kg-1),连续ig给药7 d,每天给药1次。HE染色观察丘脑病理学改变;高效液相-色谱荧光法检测丘脑谷氨酸(Glu)和γ-氨基丁酸(GABA)含量;免疫荧光单标染色法检测丘脑谷氨酸脱羧酶(GAD65)的表达;免疫荧光双标染色法检测星形胶质细胞纤维酸性蛋白(glial fibrillary acidic protein,GFAP)、缝隙连接蛋白43(Connexin43,Cx43)表达。结果:中动脉栓塞7 d大鼠丘脑部分神经元变性,和假手术组相比,丘脑Glu,GABA含量降低(P<0.05),GAD65表达减弱(P<0.05),GFAP,Cx43表达增强(P<0.01)。黄连解毒汤水提取物及3个部位均可不同程度减轻丘脑神经元变性程度。总黄酮可明显上调丘脑GAD65表达,降低GFAP,Cx43蛋白表达,差异较模型组明显(P<0.01),促进丘脑Glu,GABA含量恢复至正常水平。黄连解毒汤水提取物、总生物碱组、总环烯醚萜组大鼠丘脑Cx43蛋白表达较模型组明显上调(P<0.01),水提物组大鼠GAD65表达也较模型组明显增高(P<0.05)。黄连解毒汤水提取物、总生物碱组、总环烯醚萜组大鼠丘脑GFAP蛋白表达较模型组没有显著性差异,GABA含量明显低于假手术组(P<0.05)。结论:中动脉栓塞后,丘脑可出现神经细胞变性,递质氨基酸含量减少,星形胶质细胞异常活化,GFAP,Cx43蛋白表达异常。黄连解毒汤总黄酮可减轻脑缺血后丘脑神经细胞损伤,其作用与抑制星形胶质细胞异常活化,调节GFAP,GAD65蛋白表达关系密切。
Objective： To observe the protective effects of the Huanglian Jiedu decoction aqueous extract and its active fraction,which consists of total alkaloids,total flavonoids and total iridoid,on the thalamus of cerebral ischemia in rats. Method： The rat model of middle cerebral artery occlusion（ MCAO） was chosen. Male SD rats were randomly divided into sham-operation group,model group,aqueous extract group（ 800 mg·kg^-1）,total alkaloids group（ 44 mg·kg^-1）,total flavonoids group（ 50 mg·kg^-1） and the total iridoid group（ 80 mg·kg^-1）. The rats were administered the appropriate drugs intragastrically once a day,for 7 days after surgery. An equivalent volume of saline was given in the sham surgery and model groups. The HE staining was adopted to observe the pathological changes. Determination of Glu and γ-GABA in thalamus were detected by HPLC with fluorescence detection. The expression of GAD65 was examined with immunohistochemistry and double staining with uorescent-conjugated antibodies against GFAP and Cx43 was chosen in this study. Result： The neurons degenerated in MCAO rats after cerebral ischemia 7 d. The content of Glu,γ-GABA decreased（ P〈 0. 05）,the expression of GAD65 reduced（ P〈 0. 05） and the expression of GFAP and Cx43 increased（ P〈 0. 01） in thalamus of rats compared with sham-operation group. Huanglian Jiedu decoction aqueous extract,total alkaloids,total flavonoids and total iridoid reduced the degeneration of neurons. Total flavonoids could promote the expression of GAD65（ P〈 0. 05） and decrease the expression of GFAP and Cx43（ P〈 0. 01） in thalamus compared with model group while it could also increased the content of Glu,γ-GABA to normal levels. Compared with model group,Huanglian Jiedu decoction aqueous extract,total alkaloids and total iridoid could raise the expression of Cx43,and Huanglian Jiedu decoction aqueous extract could also increase the expression of GAD65（ P〈 0. 05）.The expression of GFAP in Huanglian Jiedu decoction aqueous extract group,total alkaloids group and total iridoid group were not different compared with model group while the content of γ-GABA decreased（ P〈 0. 05） compared with sham-operation group. Conclusion： The degeneration of nerve cells,the reduction of neurotransmitter amino acids content,the aberrant activation of astrocytes and the abnormal expression of GFAP and Cx43 will appear in thalamus of MCAO rats after ischemia. Huanglian Jiedu decoction total flavonoids could relieve the injury of nerve cell through inhibiting the abnormal activation of astrocytes and regulating the expression of GFAP and GAD65.
出处
《中国中药杂志》
CAS
CSCD
北大核心
2014年第22期4405-4410,共6页
China Journal of Chinese Materia Medica
基金
国家科技支撑计划项目(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(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)
