Effect And Mechanism On Digestive Cancer Of Xiao'ai Jiedu Prescription

Objective:Xiao'ai Jiedu prescription is an anti-cancer prescription designed by Medica master Zhou zhongying using Pathogenesis Theory of Cancerous Toxin.The efficacy of this prescription is confirmed by decades of clinical and experiment research,whereas the mechanism remains unclear.In this study,we made DEN-induced hepatocellular carcinoma model with SD rats and subcutaneous transplantation colon cancer model with CT26 mice in vivo,and chose BEL-7402,SMCC-7721,HepG2 in vitro,to investigate efficacy and mechanism Xiao'ai Jiedu prescription.This study was used to discover innovative drug.Method:1(1)70 male 270g±20g SD rat were randomly divided into DEN group(60)and normal group(10).Rats in DEN group were given 0.01%DEN contained water,and the normal group were given normal drinking water,at 4,8,12 week 3 random rats in DEN group were killed and liver were harvest to observe the cancer progression.After 13 week modeling,model rats were randomly divided into XJD high(10,26g/kg/d,i.g.),XJD mid(10,13g/kg/d,i.g.),XJD low(10,6.5/kg/d,i.g.),and model(10).XJD and model group were given normal drinking water.Rats characteristics were observed and recorded in each group during modeling progression.After 15 days treatment,all rats were killed and tissue were harvest and weighted,liver were staining with H&E method.IL-1? and IL-6 inflammation cytokines level was measured by ELISA assay,IL-1? and IL-6 expression was detected by Immunohistochemistry in rat liver.?-SMA,Bax,Bcl-x1,p21,p53,MMP2,MMP9 protein expression were detected by western assay to observe the effect on TGF-pl/Smad,TLR4/MYD88/NF-?b signaling pathway by Xiao'ai Jiedu prescription.CD68?CD206?Arg-1?Dectin-1 mRNA expression were detected by RT-PCR.(2)280g SD rats were randomly divided into XJD group(15)and normal group(5).XJD group were given 65g/kg/d XJD by intragastric administration all 10 days.After 10 days,all rats were killed,serum were collected.After 56oC water bath,30min inactivate,sterile filtered,and drug serum was prepared and stored at-80? refrigerator.Proliferation of BEL-7402,SMCC-7721,HepG2 were ascertained by MTT assay in 24,48,72h.10%,8%,5%,2%serum drug were prepared with 1640 blank media,effect of drug serum on proliferation were detected separately.We used PMA,IL-3 and IL-4 to stimulate THP-1 to differentiate into M2 macrophage.Supernatant were collected to made the M2 macrophage conditioned media.Conditioned media were used to culture SMC-7721,and XJD drug serum were added into the culture,to investigate XJD efficacy on M2-condtioned hepatocellular carcinoma cell.2.Insert block method were used to made colon cancer model,and CT26 mice were randomly divided into Veh group,OXA group,XJD group.Body weight,tumor volume,tumor weight were detected and inhibition rate was calculated.F4/80,TGF-?1,IL-6,IL-1?expression were detected by western and immunohistochemistry assay.XJD group was compared with model group and oxaliptin,to discover the XJD efficacy on colon cancer.Results:1(1)We found that weight is higher in control group than in DEN group,and weight is higher in XJD group than in DEN group.Liver weight is higher in DEN group than in control group,whereas liver weight is higher in DEN group than in XJD group.XJD inhibit the high expression in DEN-induced hepatocellular carcinoma,and shows a dose-dependent manner.Further,XJD can inhibit DEN-induce high IL-6 IL-1? expression.High concentration XJD can lower the inflammation cytokined expression.XJD can inhibit hepatic fibrosis,blocking TGF-?/SMAD,TLR4/MYD88/NF-?b pathway,inhibit macrophages M1-M2 polarization,increase apoptosis of cancer cell and MMP expression.(2)In this study,48h 10%drug serum show great inhibition on proliferation of SMC-7721,72h 5%drug serum show great inhibition on M2 conditioned SMC-7721cell.2.XJD can significantly inhibit the tumor growth,but not greater than oxaliplatin.F4 1 80,TGF-?1,IL-6 and IL-1?expression were higher in XJD group than in model group.Conclusion:Xiao'ai Jiedu prescription is positively effect in digestive system cancer.And the mechanism against hepatocellular carcinoma involves inhibition in immune escape,inflammation,Tumor invasion and metastasis,Macrophage M1 polarization,and induction of cancer Apoptosis.Whereas mechanism against colon cancer involves induction of macrophage in cancer region and enhance immunogenicity.