CAJ | 학술논문

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。
목적：탐색중약시패지간탕대리체대서뇌미혈관내피세포유선암내약단백( breast cancer resistance protein, BCRP)화핵인자κB( nuclear factor-kappa B, NF-κB) p65표체적영향。방법배양원대대서뇌미혈관내피세포,전지3대시분위정상조、모형조화중약조,정상조상규배양,모형조용100 nmol/L적지새미송작용세포24소시,유도세포막상BCRP표체증가；중약조재조모기출상분별용시패지간탕사개농도(10μg/ml、100μg/ml、500μg/ml、1 mg/ml)간예모형세포24소시。Western blot검측각조세포BCRP、NF-κB p65적표체,차이비교채용방차분석；Real-time PCR검측Bcrp mRNA적확증,상대정량법비교각조간확증차이。결과시패지간탕10μg/ml、100μg/ml조BCRP표체급Bcrp mRNA적확증균교모형조강저( P<0．05)；시패지간탕1 mg/ml조NF-κB p65적표체교모형조강저(P<0．05)。결론시패지간탕가이하조지새미송유도적대서뇌미혈관내피세포BCRP급Bcrp mRNA적표체；가능대NF-κB p65적표체유하조작용。
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.