| Objective:1.Berberine whether or not to protect intestinal mucosal permeability in colon cancer2.Colon cancer cells whether or not to have a predatory effect on intestinal polyamines in the process of colon cancer.3.Berberine whether or not to protect intestinal mucosa through polyamine mechanism in the process of colon cancer.Methods:1.Inhibitory effects of berberine on growth of transplanted tumor bearingmice（HCT116）The model of HCT116 was randomly divided into control group,model group,berberine low dose（6.25 mg/kg）,berberine middle dose（12.5 mg/kg）,berberine high dose（25 mg/kg）,and observed the daily life signs and eating conditions of nude mice.The inhibitory effect of berberine on the growth of xenografts in nude mice was evaluated by measuring the tumor volume of nude mice,measuring the tumor weight and the tumor suppressor rate after the end of the administration,and combining the weight change curve of the mice with the tumor bearing mice.2.Effects of berberine on intestinal permeability in tumor bearing mice（HCT116）The concentration of D-lactate in the plasma of tumor bearing mice,HE pathological section of intestinal mucosa,orrelation analysis of tumor weight and plasma D-lactic acid,protein expression of intestinal tight junction protein ZO1 and Occludin,all of these were used to observe the effect of berberine on intestinal permeability in tumor bearing mice.3.Protective effect of berberine on intestinal mucosa in tumor bearing mice（HCT116）（1）In order to explore the effect of berberine on Polyamine Content in HCT116 and IEC6 co culture system:The inhibitory effect of berberine on HCT116 cell proliferation was determined by CCK8,and berberine dosage and the best time of action were determined.;IEC6 cell single culture（24 h,48 h,72 h）and HCT116 cells of different time points（24 h,48 h,72 h）were co cultured with IEC6 cells and 2,and the changes of IEC6 cell permeability（FD4 concentration）were observed;High performance liquid chromatography was used to determine the content of polyamines in two cells after co culture of HCT116 cells and IEC6 cells.（2）In order to explore the effect of berberine on polyamine content,tumor polyamine content and polyamine metabolism protein expression in intestinal mucosa of HCT116 tumor bearing mice,high performance liquid.chromatography was used to determine the content of polyamines in intestinal mucosa and tumor;and WB method was used to observe the expression of tumor polyamine metabolism protein ODC,OAZ1,SSAT,C-MYC and HIF-1αafter berberine administration.Results:1.Inhibitory effects of berberine on growth of transplanted tumor bearingmice（HCT116）The tumor volume of the model group increased the fastest,compared with the model group,the tumor volume increased slowly.The tumor weight of the model group was significantly higher than that of the drug delivery group.Meanwhile,the tumor inhibition rate was 20.9%,33.7%,42.2%,respectively.The body weight of the normal group was significantly higher than that of the administration group and the model group,among which the model group was the lowest,and in the third week,the weight loss of the drug group and the model group was reduced,the cancer dystrophy may appear.2.Effects of berberine on intestinal permeability in tumor bearing mice（HCT116）The plasma D-lactate concentration in the normal group was the lowest.Compared with the normal group,the model group and the drug delivery group were all higher,and the model group was the highest.The intestinal mucosal barrier was severely damaged in the model group,compared with the model group,the intestinal mucosa was obviously restored in the treatment group.We found that plasma D-lactic acid is closely related to tumor（P<0.001）when calculating the correlation curve between plasma D-lactic acid and tumor weight.The results of the close connexin ZO1,Occludin showed that the tight connexin was down regulated in the model group,compared with the normal group,the expression of the close connexin was up after administration.3.Protective effects of berberine on intestinal mucosa in tumor bearing mice（HCT116）（1）Effects of berberine on Polyamine Content in HCT116 and IEC6 co culture system:CCK8 datas showed that berberine（0-50μM）had inhibitory effects on HCT116 cells,and the best effect was 72 hours,48 hours later.HCT116 cells were co cultured with IEC6 cells,we explored that 48h was the best co culture time by measuring the FD4 concentration of IEC6.In the determination of polyamine content in HCT116 and IEC6 cells after co culture,the result was Hco>H codrug>H single,I single>I coadministration>Ico;（2）Effects of berberine on polyamine content,tumor polyamine content and polyamine metabolic protein expression in intestinal mucosa of HCT116 tumor bearing mice:Compared with the normal group,the concentration of polyamines in the intestinal mucosa was lower than that in the normal group.Compared with the model group,the content of polyamines increased,and the content of polyamines increased with the increase of the concentration of the drug.Compared with the model group,the total polyamines decreased significantly compared with the model group,and the high dose of the administration group was statistically significant（P<0.001）.The tumor polyamine metabolic protein showed C-MYC,ODC down regulation,OAZ1 up regulation,up regulation of SSAT,and up regulation of HIF-1α.Conclusion:1.Berberine can protect intestinal mucosal permeability in colon cancer.2.Colon cancer cells have a predatory effect on intestinal polyamines in the process of colon cancer.3.Berberine can protect intestinal mucosa through polyamine mechanism in the process of colon cancer. |
