丹芪偏瘫胶囊中羚羊角用人工牛黄替代的体外研究  

Substitution of antelope horn in Danqi Piantan capsule with artificial bezoar in vitro

作　　者：王进博[1,2] 李正[1] 陈涛[1] 张艳军[2] 崔维利[1] 李进[1] 

机构地区：[1]天津中医药大学第一附属医院,天津300193 [2]天津中医药大学,天津300193

出　　处：《中国中药杂志》2015年第22期4456-4462,共7页China Journal of Chinese Materia Medica

基　　金：天津市科技创新专项资金项目(06FZZDSH00408);天津市西青区科技创新专项计划项目(XQCXZX2012-006);天津市杀手锏项目(XQKC2013-022)

摘　　要：为研究丹芪偏瘫胶囊(DPC)及DPC去羚羊角人工牛黄翻倍(DPCBD)对神经再生和血管再生的作用,初步探讨人工牛黄替代羚羊角的可能性,进行了体外培养细胞实验。本实验分为空白血清对照组、模型组、DPC组(0.306 g·kg-1·d-1,以下相同)、丹芪偏瘫胶囊去羚羊角(DPCRA)组、DPCBD组。体外培养脑微血管内皮细胞(BMEC)、星形胶质细胞(astrocytes)以及神经干细胞(neural stem cells,NSC),并将3种细胞共培养,模拟神经血管单元,用微管相关蛋白Ⅲ(β-tubulinⅢ)抗体标记神经元,用胶质纤维酸性蛋白(GFAP)标记astrocytes。酶标仪法检测BMEC乳酸脱氢酶(LDH)含量,倒置相差显微镜观察BMEC管样结构形成,显微镜下计数与BMEC黏附的白细胞个数,免疫荧光法检测β-tubulinⅢ和GFAP阳性细胞比例以及RT-PCR法检测NGF,BDNF,VEGF和VEGFr-2 mRNA的表达水平。结果显示,与模型组相比,DPC和DPCBD均可减少LDH漏出;均可促进BMEC管样结构形成;均可抑制白细胞与BMEC黏附;均可增加β-tubulinⅢ阳性细胞分化比例(P<0.01),降低GFAP阳性细胞比例(P<0.01);均能在一定程度上增加共培养细胞NGF,BDNF,VEGF和VEGFr-2 mRNA的表达,其中对NGF和VEGF mRNA表达水平的影响具有显著性差异(P<0.05),且2组疗效相当。DPCRA组对各指标的影响明显不如DPCBD和DPC组。DPCBD与DPC对神经再生和血管再生的作用疗效相当,提示丹芪偏瘫胶囊中羚羊角可用人工牛黄进行替代。The in vitro cell culture experiment was conducted to study the effect of Danqi Piantan capsule（ DPC） and DPC dislodge the antelope horn with artificial bezoar double（ DPCBD） on nerve regeneration and blood vessel regeneration and preliminarily investigate the possibility of substituting antelope horn in DPC with artificial bezoar. In this experiment,rats were randomly divided into 5groups： the blank serum control group,the model group,DPC groups（ 0. 306 g·kg-1·d-1,the same below）,DPC remove of antelope horn（ DPCRA） groups and DPCBD groups. Brain microvascular endothelial cells cultured in vitro（ BMEC）,astrocytes and neural stem cells（ NSC） were co-cultured to simulate neurovascular unit,label neurons with microtubule associated protein III（ β-tubulin III） antibody and lable astrocytes with glial fibrillary acidicprotein（ GFAP）. ELISA was used for the detection of the content of BMEC lactate dehydrogenase instrument method（ LDH）,the inverted phase contrastmicroscope was adopted to observe the formation of BMEC tube like structure,the number of leukocytes and leukocytes adherent to BMEC were counted under the microscope,the expression levels of β-tubulin III and the ratio of GFAP positive cells was detected with inimmunofluorescence,and RT-PCR method was used to detect NGF,BDNF,VEGF and VEGFr-2 mRNA. According to the result,compared with the model group,both DPC and DPCBD can reduce LDH leakage,promote the formation of BMEC tube like structure,inhibit leukocytes and their adhesion to BMEC,increase theβ-tubulin III positive cell differentiation proportion（ P 0. 01）,reduce the proportion of GFAP positive cells（ P 0. 01）,increase the expressions of co-cultured NGF,VEGF,BDNF and VEGFr-2 mRNA to a certain extent,with the most significant difference on NGF and VEGF mRNA expressions（ P 0. 05） and the same efficacy in both groups. DPCRA groups showed less impact on all indexes than that of DPCBD and DPC groups. The same efficacy of DPCBD and DPC on nerve regeneration

关 键 词：丹芪偏瘫胶囊 丹芪偏瘫胶囊去羚羊角人工牛黄翻倍 脑微血管内皮细胞 神经干细胞 神经再生 血管再生 

分 类 号：R285.5[医药卫生—中药学]