Antitumor Activity Of Jianpi-Qingre-Chushi Method In CRC Tumor Bearing Mice With Dampness-heat Syndrome And Its Effect On Gut Microbiota

Objective: Objective antitumor activity of jianpi-qingre-chushi method(compound traditional chinese medicine)and its effect on gut microbiota of tumor bearing mice with dampness-heat syndrome by establishing mouse model of colorectal cancer with dampness-heat syndrome,reveal its anti-tumor mechanism,and provide the basis for clinical medication.Methods: 1)Grouping:select 40 BALB/c male mice(4-6 weeks old),after adaptive feeding for one week,the mice were optionally falled into 5organizes: blank organize(group Con),model organize(group Model),high-dose organize of traditional chinese medicine(group ZQYJ-H),medium-dose organize of traditional chinese medicine(group ZQYJ-M)and low-dose organize of traditional chinese medicine(group ZQYJ-L),with 8 mice in each group.2)Modeling:in order to establish the dampness-heat model,mice in traditional chinese medicine group and model group were fed with honey water(0.2g/m L)and gavaged with Beijing Hongxing Erguotou(52°)(0.1m L),pork fat(0.1m L)for 7 days.3)Gavaging:after the subcutaneous tumor grew,the mice in traditional chinese medicine group were given high(10g/kg),medium(5g/kg)and low(2.5g/kg)doses of ZQYJ(zhenqi-liujun anti-cancer decoction combined with yi-jiajian-zhengqi powder)by gavage(0.4m L),and the mice in blank group and model group were given normal saline by gavage(0.4m L).The mice were killed 21 days after taking the drug.4)Recording:during the experiment,the body weight,mental state and feces condition of mice were recorded.5)Testing:after the mice were killed,the tumor size and pathological changes were observed.We detected the expression of Akt,PI3 K,Bcl2 and Caspase3 proteins by Western blot,the m RNA expressions of AKT,PI3 K,Bcl2and Caspase3 were detected by Real-time PCR and the gut microbiota was detected by 16 Sr RNA.Results: 1)During the administration period,the general condition of mice in traditional chinese medicine group was better than that in the blank group and model group.Compared with model group,the body weight of mice in group ZQYJ-H,ZQYJ-M increased(P < 0.05).Compared with the blank group and the model group,the tumor in the traditional chinese medicine group grew slowly and the volume was smaller(P < 0.05).The subcutaneous tumor weight in ZQYJ-H,ZQYJ-M group was decreased compared with that in blank group.Compared with blank group,the subcutaneous tumor weight of model group and ZQYJ-L group was not reduced(P > 0.05).The subcutaneous tumor weight in group ZQYJ-H,ZQYJ-M were inferior of that in group model(P < 0.05).There was no difference in tumor weight between group ZQYJ-L and group model(P > 0.05).The tumor inhibition rate of ZQYJ-H group was 44.42%,that of ZQYJ-M group was 26.34%,and that of ZQYJ-L group was 13.83%.Compared with blank group and model organize,the traditional chinese medicine group had more necrosis.The necrosis of tumor tissue was the most serious in group ZQYJ-H,the second in group ZQYJ-H,and the least in group ZQYJ-L.3)The protein expressions of AKT,PI3 K,BCL2 and Caspase3:there was no difference in AKT protein expression in each organize(P > 0.05).The expression of PI3 K protein in group ZQYJ-H and ZQYJ-M was inferior of that in blank group and model group(P < 0.05).The expression of Bcl2 protein in group ZQYJ-H,ZQYJ-M and ZQYJ-L was lower than that in blank group and model group(P < 0.05).Compared with blank group and model group,the expression level of Caspase3 protein in organize ZQYJ-H and ZQYJ-M increased(P < 0.05).3)The expression of AKT,PI3 K,BCL2 and Caspase3 m RNA: the expression of AKT1 m RNA in group ZQYJ-H and ZQYJ-M was decreased contrasted with organize Con(P < 0.05),and the express of AKT1 m RNA in organize ZQYJ-H,ZQYJ-M and ZQYJ-L was decreased contrasted with model group(P < 0.05).The expression of PI3 K m RNA in organize ZQYJ-H and ZQYJ-M was inferior of that in blank group and model group(P <0.05).The expression of Bcl2 m RNA in group ZQYJ-H and ZQYJ-M was inferior of that in blank group and model group(P < 0.05).The expression of Caspase 3 m RNA in organize ZQYJ-H and ZQYJ-M was taller than that in blank group and model group(P < 0.05),and the expression of Caspase3 m RNA in organize ZQYJ-L was taller than that in organize Con(P <0.05).4)Zhenqi-liujun anti-cancer decoction combined with yi-jiajian zhengqi powder can change the gut microbiota of tumor bearing mice,increase the number of lactobacillaceae and lachnospiraceae in the gut microbiota of tumor bearing mice with colorectal cancer,reduce the number of muribaculaceae and staphylococcaceae,promote the growth of prevotellaceae and lactobacillaceae in the gut microbiota of tumor bearing mice with dampness-heat syndrome,and inhibit the growth of staphylococcaceae.Conclusion:1)Zhenqi-liujun anti-cancer decoction combined with yi-jiajian-zhengqi powder can increase the body weight of tumor-bearing mice,enhance the food intake of tumor-bearing mice,improve the mental state of tumor-bearing mice,reduce the weight of subcutaneous tumors,and promote the necrosis of colorectal cancer tissue.2)Zhenqi-liujun anti-cancer decoction combined with yi-jiajian-zhengqi powder can inhibit the activation of PI3 K /AKT pathway to lay the antitumor effct.3)Zhenqi-liujun anti-cancer decoction combined with yi-jiajian-zhengqi powder can change the gut microbiotaa of tumor bearing mice and promote the growth of probiotics.