不同方法提取化瘀消癥复方对HTR-8/SVneo细胞凋亡的影响

Effect on HTR-8/SVneo Apoptosis by Different Extractions of Compound Huayu Xiaozheng Formula

目的:探讨不同方法提取化瘀消癥复方(丹参、桃仁、赤芍、天花粉和紫草)对人源妊娠滋养细胞系HTR-8/SVneo细胞凋亡的影响。方法:采用CCK8法检测经不同方法提取的化瘀消癥复方(A水提、B醇提、C醇提后药渣水提、D醇提+水提)对HTR-8/SVneo细胞增殖的影响;流式细胞术检测48 h后细胞凋亡情况;设甲氨蝶呤组为阳性对照组,未经药物处理的细胞为阴性对照组。结果:不同提取方法的中药复方对HTR-8/SVneo细胞作用后对其增殖均有抑制作用,其中A组抑制率大于B、C、D组(P<0.05);其凋亡率分别为(24.37±3.24)%(A组),(12.83±0.37)%(B组),(11.70±0.48)%(C组),(14.37±0.51)%(D组),(18.05±1.27)%(MTX组),与阴性对照组(7.057±0.76)%相比,各组凋亡率均明显增加(P<0.01);其中A组凋亡率高于MTX组、B组、C组、D组(P<0.01)。结论:不同提取方法提取化瘀消癥复方对HTR-8/SVneo细胞凋亡的影响,以水提组最明显,可将水提法作为化瘀消癥复方后续研究的中药提取方法。

Objective：To study the effect on anthropogenic trophocyte HTR-8/SVneo apoptosis by different extractions of Compound Huayu Xiaozheng Formula which includes Danshen（Salvia miltiorrhiza）,Taoren（Semen persicae）,Chishao（Radix paeoniae rubra）,Tianhuafen（Radix trichosanthis） and Zicao（Lithospermum erythrorhizon）.Methods： Determined by CCK8 method and flow cytometry,we detected the proliferation and apoptosis of anthropogenic trophocyte HTR-8/SVneo after cultivating with different extractions of Compound Huayu Xiaozheng Formula（group A-water extraction,group B-alcohol extraction,group C-alcohol extraction after water extraction,group D-alcohol extraction and water extraction）.Result：The inhibition ratio by water extraction（group A） of Compound Huayu Xiaozheng Formula was higher than that of other groups（P〈0.05）.The apoptotic indexes were（24.37 ± 3.24） %（group A）,（12.83 ±0.37） %（group B）,（11.70 ± 0.48） %（group C）,（14.37 ± 0.51） %（group D） and（18.05 ± 1.27） %（group MTX）.The apoptotic indexes of group A were higher than that of other groups（P〈0.01）.Conclusion： The water extraction of Compound Huayu Xiaozheng Formula had better apoptotic effect on HTR-8/SVneo cells than other extraction methods.Water extraction method can be used as the extraction method of this formula in further studies.