Studythe Purification Of Qichan Koufu Jiechang Baxiang Tablets Effective Components And Its Function Of Anti-colon Carcinoma In Vitro

Objective Research the purification of Qichan Koufu Jiechang Baxiang Tablets effective components,further research its inhibitory effect on colon cancer cell CT-26,and effect of apoptosis related gene Bax and Bcl-2,angiogenesis related factor VEGF and HIF-la expressi-on,to investigate its function of anti-colon carcinoma in vitro.For in vivo research provides the experimental basis,for the Qichan Koufu Jiechang Baxiang Tablets effective components used for clinical prevention,treatment and rehabilitation of colon cancer provide theoretical basis.Method purification of total saponins of astragalus and Matrine from Sophorae with macrop-orous adsorption resin,purification of cinobufagin and resibufogenin with silicagel column the inhibitory effect on colon cancer cell CT-26 of Qichan Koufu Jiechang Baxiang Tablets ef-fective components were evaluated by MTT colorimetric method,Real time RT-PCR method to detect the effect of Qichan Koufu Jiechang Baxiang Tablets effective components on apopt-os related gene Bax,Bcl-2 expression;Westem blot to detect the effect of Qichan Koufu Jiech-ang Baxiang Tablets effective components expression on VEGF and HIF-1? expressionResults The technology optimization of purification for total saponins of astragalus is Selec-ting the AB-8 adsorption resin,the sample amount is 220 mL(crude drug concentration 0.2 g/mL)elution with distilled water to wash to no molish reaction,then 0.1%NaoH and 20%ethanol elution in removing impurity,70%ethanol elution,elution amount to 5BV.The technol-ology optimization of purification for Matrine from Sophorae is Selecting the D101 adsorpti-on resin,the sample amount is 120 mL(crude drug concentration 0.2 g/mL),distilled water and 20%ethanol elution in removing impurity,then 80%ethanol elution,elution amount to 5BV The technology optimization of purification for cinobufagin and resibufogenin is process for diameter ratio for 2:14,ample amount for l0g,elution solvent dosage is 160 mL.Qichan Koufu Jiechang Baxiang Tablets effective components could inhibit colon cancer CT-26 cells,Qichan Koufu Jiechang Baxiang Tablets effective components acting on CT-26 cells 24h,48h,72h,IC50 were 95.04?49.84?23.67 ?g/mL.Qichan Koufu Jiechang Baxiang Tablets effective components its inhibition rate of colon CT-26 cells with time and dose dependent.Compared with the 5Fu group Qichan Koufu Jiechang Baxiang Tablets effective components 300.00 ?g/mL group no Statistically difference(P>0.05).Qichan Koufu Jiechang Baxiang Tablets effective components acting on CT-26 cells 24h,48h,72h,compared with the blank group,Bax gene expression was enhanced,and there was statistically difference(P<0.01,P<0.05),Bcl-2 gene expression was reducedand there was significant difference(P<0.01).Compared with the 5Fu group,Bax gene expression have significant differences(P<0.01),Bcl-2 gene expression have statistically difference(P<0.01,P<0.05).Bax,Bcl-2 gene expression with time and dose dependent in Qichan Koufu Jiechang Baxiang Tablets effective components groups.After 48h,compared with the blank group,VEGF and HIF-1a expression was reduced,and there was significant differences(P<0.01).compared with the 5Fu group,75.00 ?g/mL group VEGF and HIF-1a expression was no Statistically difference(P>0.05).VEGF and HIF-1a expression with dose dependent in Qichan Koufu Jiechang Baxiang Tablets effective components groups and there was statistically difference(P<0.01,P<0.05).Conclusion Macroporous adsorption resin method is applicable to the purification of total saponins of astragalus and Matrine from Sophorae.silica gel column is suitable for purificati on of cinobufagin and resibufogenin.After purification of total saponins of astragal us,Matrine from Sophorae,cinobufagin and resibufogenin,their purity were improved.Purification of Qichan Koufu Jiechang Baxiang Tablets effective components could inhibit colon cancer CT-26 cells,could upregulation the apoptosis gene Bax,and downregulation Bcl-2,could promote apoptosis of colon CT-26.could inhibit the expression of VEGF and HIF-1a has inhibitory effect on tumor angiogenesis.Qichan Koufu Jiechang Baxiang Tablets effective components can anti-colon carcinoma in vitro.