| Objective:The anti-hepatocellular carcinoma effect of YFBS was studied.In addition,the compatibility of YFBS in liver cancer was studied from the perspective of pharmacological effect and chemical composition.Methods:MTT method was used to evaluate the inhibitory effect of water extract and serum of YFBS on the proliferation of hepatocellular carcinoma cells in vitro.Hepal-6 hepatocellular carcinoma model was administered by intragastric administration for 2 weeks to extract the tumor,and the overall effect of the drug was evaluated by tumor inhibition rate.The three drugs of YFBS were divided into the single drug group and pair-wise fomulation group.Hepal-6 hepatocellular carcinoma cell proliferation assay was performed in vitro,and its contribution to the effect was evaluated by MTT assay.HPLC was used to detect the fingerprints,the attribution of common peak and the source of anti-tumor components of YFBS decoction.The chromatographic conditions were as follows:the chromatographic column was Agilent ZORBOX Extend-C18 column(250 mm×4.6 mm,5 m),the mobile phase was acetonitrile-0.05%phosphoric acid solution(pH2.5),and the elution method was gradient elution.The flow rate of 1.0 mL/min was set,with each injection of 10μL,and the detection was conducted at the wavelength of 230 im and the column temperature was 30℃.The optimal compatibility ratio of prescriptions was investigated in the mice model of liver cancer.The method of uniform design U9(94)was used and the tumor inhibition rate was used as the evaluation index.Results:In vitro experiments,compared with the control group,YFBS could significantly inhibit the proliferation of Hepal-6,HepG-2 and SMCC-7721 cells in time and dose dependence.Moreover,YFBS was more sensitive to Hepal-6 hepatocellular carcinoma cells at the same concentration,and the inhibition rate was higher.The OD number of hepal-6 hepatocellular carcinoma cells under the action of 5%,10%,15%,and 20%of YFBS containing serum decreased compared with that in the blank group,and the drug containing serum inhibited the proliferation of cells and was dose-related.In vivo,compared with the model control group(1.83±0.72)g,the tumor weight in the 7.735 and 15.47 g/kg YFBS group was(1.21±0.33)g and(0.79±0.33)g,respectively.The tumor inhibition rates were(33.78±17.85)%and(56.95±18.17)%,respectively.In the dismantling experiment,it was found that the dahurian patrinia herb group in the single drug group had the strongest in low effective concentration,followed by the aconite group,and the coix seed group was the weakest.In the pair-wise fomulation group,the dahurian patrinia herb+aconite group showed the strongest inhibitory effect on the proliferation of Hepal-6 cells at low concentration,followed by the coix seed+dahurian patrinia herb group,and the coix seed+aconite group was the weakest.The fingerprinting of the decoction of YFBS was established by HPLC-DAD and a total of 51 chromatographic peaks were identified.Eight peaks were identified,including Chlorogenic acid(9),Caffeic acid(11),Rutin(13),Isochlorogenic acid B(36),Isochlorogenic acid A(37),Benzoylmesaconine(38),Benzoylaconitine(41),Quercetin(46).And three peaks were derived from coix seed(peaks 2,28,50).Twelve peaks were derived from aconite(peaks 4,5,7,8,10,14,20,25,35,38,41,51).The 42 peaks were derived from the dahurian patrinia herb(peaks 1,3,6-19,21-27,29-37,39-40,42-49).10 batches of YFBS were studied,and the similarity was above 0.996.Chlorogenic acid,Caffeic acid,Rutin,Isochlorogenic acid B,Isochlorogenic acid A,Quercetin were derived from the dahurian patrinia herb;Benzoylmesaconine and Benzoylaconitine were derived from the aconite in the decoction of YFBS.The partial least squares quadratic polynomial regression analysis was carried out for the 9 groups of data obtained from uniform design with the tumor-suppressing rate as the y value,and the equation was obtained as follows:y=37.5753171-0.203900x1-1.008153x2-3.201963x3+0.005349x1*x1+0.072443x2*x2+0.133768x3*x3-0.021211x;*x2-0.016679x1*x3+0.085996x2*x3(R2=0.7924).When the tumor inhibition rate was the highest,the equation calculated the best compatibility ratio of coix seed:aconite:dahurian patrinia herb=5:9:11.Through verification experiments,it was found that the optimized ratio of YFBS had better inhibitory effect on liver cancer than that of the original formula.Conclusion:YFBS had anti-hepatocellular carcinoma effect in vitro and in vivo.YFBS in the anti-hepatocellular carcinoma effect of the combination of junchen corresponding to jingfang changed.The dahurian patrinia herb was the medicine of the emperor.The optimal compatibility ratio was coix seed:aconitae:dahurian patrinia herb=5:9:11,and the inhibitory effect of liver cancer tumor was better than the original ratio. |
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