Anti-tumor Effects Of Compatibility,prescription Tcd Using Median-effect Principle

Objective:The present study was designed to investigate the effect of Radix Aconiti Praeparata in combination with Bulbus Fritillaria and every herbs of Siwu decoction on tumor under different drug dosage and combination ratio,are observed in vitro.Median effect principle offers quantitative definition for the interaction between them.Explore the relevant mechanisms and targets.Methods:1.Human breast cancer MDA-MB-231 cells were seeded in culture plates.Set up a system of different concentrations of Radix Aconiti Praeparata and Bulbus Fritillaria single-use group,combined-use group,as well as compatibility by 1:1,2:1,1:2 proportion and negative control group.Cell morphological changes were observed under light microscope.The antiproliferative activity was determined by trypan blue staining method.Median effect principle is used to evaluate both of them with or without joint synergies.Cell-apoptosis was investigated using flow cytometry.2.Human cervical cancer Hela cells were selected.The antiproliferative activity of every drug in Siwu decoction was determined by trypan blue staining method.The effect of positive drug in combination with other drugs on Hela cells under different drug dosage and combination ratio,are observed.Detect cell proliferation inhibition rate by trypan blue staining method.Median effect principle is used to evaluate both of them with or without joint synergies.Cell-apoptosis and cell-cycle distribution were investigated using flow cytometry for the synergy experimental group.Results:1.The effect of Bulbus Fritillaria single on MDA-MB-231 cells are significant inhibition of proliferation and cytotoxicity.When its concentration is from 31.25?g/mL to 500?g/mL,the inhibitory rate was greater than 50%.There was no obvious inhibition on MDA-MB-231 cells with Radix Aconiti Praeparata alone.2.Radix Aconiti Praeparata acted synergistically with Bulbus Fritillaria in inhibiting tumor cell growth in compatibility of 1:1 and 1:2.The drug dose is 15.625-500?g/mL,and 15.625-500?g/mL +31.25-1000?g/mL to achieve synergies.3.Apoptosis acts as the target for the anti-tumor effect of Bulbus Fritillaria.The combination of Bulbus Fritillaria 62.5?g/mL and Radix Aconiti Praeparata 62.5?g/mL induces apoptosis of MDA-MB-231 cell synergistically.4.Radix Paeoniae Alba equivalent to the dose of 9g crude drug could inhibite proliferation of Hela,and the inhibition rates were 28.60%.The same dose of other drugs of Siwu decoction has no significant inhibitory effect on Hela cells.5.Radix Paeoniae Alba acted synergistically with Radix Angelicae Sinensis in inhibiting Hela cell growth in compatibility of 1:1,1:2 and 2:1,especially the 1:2 ratios whose drug dose is Radix Paeoniae Alba 31.25-250?g/mL,Radix Angelicae Sinensis 15.625-125?g/mL.Radix Aconiti Praeparata acted antagonisticly with Rhizoma Chuanxiong or Radix Rehmanniae in compatibility of 1:1,1:2 and 2:1.6.Radix Paeoniae Alba 250?g/mL in combination with Radix Angelicae Sinensis 125?g/mL group failed to synergistic induction of apoptosis in Hela cells.7.Hela cells of Radix Paeoniae Alba 250?g/mL in combination with Radix Angelicae Sinensis 125?g/mL group were arrested in S phase increased.Protein expression of CyclinDl was reduced compared with control group(P<0.05).Conclusions:1.Bulbus Fritillaria could inhibite proliferation of MDA-MB-231 cells.2.Radix Aconiti Praeparata acted synergistically with Bulbus Fritillaria in inhibiting tumor cell growth in compatibility of 1:1 and 1:2.Apoptosis acts as the synergistic target of Bulbus Fritillaria and Radix Aconiti Praeparata.3.Radix Paeoniae Alba alone could inhibite proliferation of Hela.4.Radix Paeoniae Alba acted synergistically with Radix Angelicae Sinensis in inhibiting Hela cell growth in compatibility of 1:1,1:2 and 2:1,especially the 1:2.5.Radix Paeoniae Alba in combination with Radix Angelicae Sinensis could reduce protein expression of CyclinDl and arrest Hela cells cycle in S stage.