摘要
目的对溪黄草抗乙肝抗肿瘤活性部位进行初步筛选,为进一步分离活性成分奠定基础。方法首先在HepG2.2.15细胞模型上对三种不同方法萃取所得的溪黄草提取物进行抗乙肝活性筛选,以用于进一步分析提纯。用MTT实验检测细胞毒性,ELISA和实时定量PCR检测细胞上清液中HBsAg、HBeAg的分泌及乙肝病毒DNA含量。然后,用硅胶柱色谱法进一步提取分离活性最大的提取物,用MTT和ELISA检测所分离各组分的抗乙肝活性。最后,在MCF-7,BGC-823和HepG2细胞中检测细胞毒性最大的三个组分的抗肿瘤活性。结果乙酸乙酯萃取分离所得的提取物抗乙肝活性最大,可显著降低HepG2.2.15细胞的HBsAg、HBeAg抗原的分泌,并抑制乙肝病毒DNA的复制,被用来进一步分离提纯。从乙酸乙酯提取物中分离出14个组分,其中A3和A5组分对HBsAg有较好的抑制作用,A9组分对HBeAg有较好的抑制作用,并均存在明显的量效关系,且治疗指数均比乙酸乙酯提取物的要高。A6,A7和A11组分细胞毒性大,对不同的肿瘤细胞有不同的抑制活性。结论溪黄草的乙酸乙酯提取物及其分离成分具有很强的抑制乙肝病毒复制的作用,从而具有很好的抗乙肝病毒活性,进一步提取分离有利于有效成分的纯化而具更强的活性。此外,一些分离成分对不同的肿瘤细胞还有很高的细胞毒性,具一定的抗肿瘤作用。本实验为溪黄草在临床的使用及其作为潜在的高效抗乙肝和肿瘤药物的进一步研究开发工作提供了理论基础。
【Objective】 To screen and investigate the anti-HBV and anti-tumor activities of separated compounds from Rabdosia serra（Maxim.） Hara for the further isolation of its active entities.【Methods】 Three extracts from Rabdosia serra by using different solvents（petroleum ether,acetidin or butyl alcohol） were prepared and analyzed of their anti-HBV activities in HepG2.2.15 cells before further separation.The cytotoxicity of each extract was tested by MTT assay.The levels of HBsAg,HBeAg and HBV DNA in supernatants from the culture of HepG2.2.15 cells were detected by enzyme-linked immunosorbent assay（ELISA） and real-time quantitative polymerase chain reaction（PCR）.The most effective extract was then separated and the anti-HBV activities of separated compounds were tested by MTT and ELISA.The anti-tumor effects of three compounds with high cytotoxicity were identified using MCF-7,BGC-823 and HepG2 cells.【Results】 The acetidin extract C2,which was then further separated,had the most effective anti-HBV activity as shown by significant reductions in the secretion of HBsAg and HBeAg and the amount of HBV DNA.There were 14 compounds separated from C2,among which A3 and A5 markedly inhibited the HBsAg secretion while A9 inhibited the HBeAg secretion in a dose-dependent manner.Their TIs were all higher than that of C2.Moreover,the compounds A6,A7 and A11 had different anti-tumor activities in different tumor cells.【Conclusions】 The acetidin extract from Rabdosia serra and its effective separated compounds have anti-HBV activities by strongly inhibiting HBV replication.The further separation and purification can strengthen their anti-HBV effects.Meanwhile,some compounds have high cytotoxicity in different tumor cells.Our study provides theoretical evidences for the development of potential and efficient drugs from Rabdosia serra and its clinical use against HBV and tumor.
出处
《中国现代医学杂志》
CAS
CSCD
北大核心
2011年第12期1449-1456,1475,共9页
China Journal of Modern Medicine
基金
Changsha Science and Technology Plan(K0902033-31)
Hunan Science and Technology Plan(2009FJ3209)
