至真方含药血清对人大肠癌多药耐药细胞HCT-8/VCR内核因子κB活性及P-糖蛋白表达与功能的抑制作用  

Medicated serum prepared with Chinese herbal medicine Zhizhen Recipe down-regulates activity of nuclear factor-κB and expression of P-glycoprotein in human colorectal cancer multidrug-resistant cell line HCT-8/VCR

作　　者：蔡松柏[1] 张旭[1] 陈志霞[1] 孔令春[1] 王松坡[1] 

机构地区：[1]上海交通大学第一人民医院中医科,上海200080

出　　处：《中西医结合学报》2011年第12期1353-1359,共7页Journal of Chinese Integrative Medicine

基　　金：上海市中医胃病临床优势专科资助项目(No.2008YSZK007)

摘　　要：目的:探讨至真方是否能够调节核因子κB(nuclear factor-κB,NF-κB)并影响P-糖蛋白(P-glycoprotein,P-gp)的表达与功能。方法:细胞增殖-毒性检验(cell counting kit-8,CCK-8)法鉴定HCT-8/VCR细胞株的多药耐药性。分别用8%、16%、32%体积分数的至真方含药血清干预人大肠癌敏感细胞株HCT-8与多药耐药细胞株HCT-8/VCR,时间为24h。CCK-8法检测细胞的存活率以筛选含药血清的实验浓度,酶联免疫吸附测定法检测细胞核内NF-κB的转录活性,激光共聚焦显微镜检测P-gp在细胞内的分布,流式细胞仪测定细胞内罗丹明123(rhodamine 123,Rh123)的平均荧光强度。结果:HCT-8/VCR含药血清组NF-κB活性较HCT-8/VCR阴性对照组明显降低(P<0.01),在一定范围内与浓度呈负相关。P-gp逐渐弥漫分布于细胞质和细胞核,并出现核固缩、核裂解,同时平均荧光强度呈浓度梯度下降(P<0.01)。在Rh123蓄积实验中,经8%、16%、32%含药血清作用后,HCT-8/VCR细胞波峰右移,细胞内荧光强度明显增强(P<0.01)。结论:至真方含药血清能抑制HCT-8/VCR细胞内NF-κB的活性,降低P-gp的表达与功能。Objective： To investigate the effects of medicated serum prepared with Chinese herbal medicine Zhizhen Recipe （ZZR） on activity of nuclear factor-κB （NF-κB） and expression and function of P-glycoprotein （P-gp） in human colorectal cancer multidrug-resistant cell line HCT-8/VCR. Methods： The multidrug resistance of HCT-8/VCR cells was detected by cell counting kit-8 method, and the experimental concentrations of ZZR-medicated serum were determined by the same way. HCT-8 and HCT-8/ VCR cells were treated with ZZR-medicated serum of medium dose for 24 h. The activity of NF-κB was determined by enzyme-linked immunosorbent assay. The intracellular distribution of P-gp was detected by laser scanning confocal microscopy, and the mean fluorescence intensity of rhodamine 123 was detected by flow cytometry.Results： ZZR-medicated sera with volume fraction of 8%, 16% and 32% of medium dose were confirmed as the experimental sera. Compared with the untreated group, NF-κB activities of the ZZR-medicated serum groups （ZZR-medicated serum with volume fraction of 8%, 16% and 32% of medium dose） were obviously down-regulated （P 〈0. 01）, which had a negative correlation with the concentrations. After interfering HCT-8/VCR with ZZR-medicated serum of different concentrations for 24 h, P-gp in HCT-8/VCR transmitted gradually from cell membrane to cytoplasm and nuclei. Nuclei became pyknotic and cracking. Compared with the untreated group, the mean fluorescence intensities of ZZR-medicated serum groups declined with concentration gradients （P〈0.01）. The efflux of intracellular rhodamine 123 decreased, the wave crest shifted to right, and the intracellular fluorescence intensity strengthened （P〈0.01）. Conclusion： ZZR-medicated sera of experimental concentrations down-regulate activity of NF-κB and expression and function of P-gp in human colorectal cancer muttidrug-resistant cell line HCT-8/VCR and the effect is related to the concentrations.

关 键 词：结直肠肿瘤 P糖蛋白 NF-κB 复方 

分 类 号：R285.5[医药卫生—中药学]