柴贝止痫汤影响大鼠脑微血管内皮细胞乳腺癌耐药蛋白、核因子p65表达的研究  

Role of Chaibei Zhixian Decoction on regulating the expression of BCRP and NF-κB p65 in rat brain microvascular endothelial cells

作　　者：鄢泽然[1] 张青[1] 王潇慧[1] 王越[2] 聂莉媛 刘金民[1] 

机构地区：[1]北京中医药大学东方医院脑病科,博士研究生100078 [2]天津中医药大学第二附属医院急诊科

出　　处：《环球中医药》2014年第4期241-246,共6页Global Traditional Chinese Medicine

基　　金：国家自然科学基金(81072771);教育部博士学科点专项科研基金(20120013110011);北京中医药大学科研创新团队资助项目(2011-CXTD-23)

摘　　要：目的探索中药柴贝止痫汤对离体大鼠脑微血管内皮细胞乳腺癌耐药蛋白(breast cancer resistance protein,BCRP)和核因子κB(nuclear factor-kappa B,NF-κB)p65表达的影响。方法培养原代大鼠脑微血管内皮细胞,传至3代时分为正常组、模型组和中药组,正常组常规培养,模型组用100 nmol/L的地塞米松作用细胞24小时,诱导细胞膜上BCRP表达增加;中药组在造模基础上分别用柴贝止痫汤四个浓度(10μg/ml、100μg/ml、500μg/ml、1 mg/ml)干预模型细胞24小时。Western blot检测各组细胞BCRP、NF-κB p65的表达,差异比较采用方差分析;Real-time PCR检测Bcrp mRNA的扩增,相对定量法比较各组间扩增差异。结果柴贝止痫汤10μg/ml、100μg/ml组BCRP表达及Bcrp mRNA的扩增均较模型组降低(P<0.05);柴贝止痫汤1 mg/ml组NF-κB p65的表达较模型组降低(P<0.05)。结论柴贝止痫汤可以下调地塞米松诱导的大鼠脑微血管内皮细胞BCRP及Bcrp mRNA的表达;可能对NF-κB p65的表达有下调作用。Objective To observe the effects of Chaibei Zhixian Decoction on regulating the ex-pression of breast cancer resistance protein （ BCRP） and nuclear factor-kappaB （ NF-κB） p65 in rat brain microvascular endothelial cells. Methods Primary culture of brain microvascular endothelial cells were prepared from rats. The 3rd passage cells were exposed to dexamethasone （100 nmol/L） for 24h to up-reg-ulate the expression of BCRP. After dexamethasone administration, the cells were treated with increasing concentrations （10 μg/ml, 100 μg/ml, 500 μg/ml and 1 mg/ml） of Chaibei Zhixian Decoction for 24h. BCRP and NF-κB p65 protein expression were assessed by Western blot analysis and data were measured with analysis of variance. Real-time PCR was performed for Bcrp mRNA amplification and differences a-mong groups were assessed by relative quantitative assay. Results Exposure to Chaibei Zhixian Decoction （10 μg/ml, 100 μg/ml） resulted in significant decrease in protein and mRNA levels of BCRP （ P 〈0. 05）;While BCRP was down-regulated, NF-κB p65 expression was decreased by Chaibei Zhixian Decoction （1 mg/ml） （P〈0. 05）. Conclusion Chaibei Zhixian Decoction down-regulate both protein and mRNA expression of BCRP and probably decrease the levels of NF-κB p65 in rat brain microvascular endothelial cells.

关 键 词：柴贝止痫汤 乳腺癌耐药蛋白 大鼠脑微血管内皮细胞 难治性癫痫 

分 类 号：R277.7[医药卫生—中医学]