Study On The Intervention Mechanism Of Guzheng Xiaoai Decoction On Colorectal Cancer

Objective:To observe the effect of Guzheng Xiaoai Decoction containing serum on HT-29 cells and the anti-tumor effect of Guzheng Xiaoai Decoction on nude mice bearing HT-29 cell tumors.From the cell experiment and animal experiment,explore the specific anti-cancer mechanism of Guzheng Xiaoai Decoction,and provide detailed treatment basis for the clinical practice of Guzheng Xiaoai Decoction.Methods:1.Cell experiment of Guzheng Xiaoai Decoction:After treating human colorectal cancer LOVO cells,HT-29 cells,and HCT-116 cells with sterile serum containing Guzheng Xiaoai Decoction,observe the cell morphology and use CCK-8 method to know the inhibitory effect.For the inhibitory effect of this cell line,the most sensitive cell line is selected.After that,the the most sensitive were grouped,including:the low-dose group contained 10%Guzheng Xiaoai Decoction serum,the middle-dose group contained 20%Guzheng Xiaoai Decoction serum,and the high-dose group contained 30%Guzheng Xiaoai Decoction and blank control group.Scratch experiments were performed on the most sensitive cells in different groups,and Western blot was used to observe the expression of NF-κB pathway related proteins and apoptosis proteins.2.Animal experiment of Guzheng Xiaoai Decoction:A nude mouse model of tumor-bearing the most sensitive cells was established for 6-week-old nude mice.This study was divided into:the model group,8 tumor-bearing nude mice were given normal saline 0.2ml/day.8 tumor-bearing nude mice in the low-dose Guzheng Xiaoai Decoction group,converted to nude mice Guzheng Xiaoai Decoction based on 5g/kg decoction content,gavage twice a day;8 nude mice bearing tumors in the medium-dose Guzheng Xiaoai Decoction group,with lOg/kg as the standard,gavage twice a day;high-dose Guzheng Xiaoai decoction group tumor-bearing 8 nude mice,with 20g/kg as the standard,gavage twice a day.After 14 days of intervention,the tumor-bearing nude mice were sacrificed,the tumors were removed,and HE staining was used for pathological observation.Perform immunohistochemical staining on the removed tumor tissues to observe the differences in Bcl-2 and Bax proteins in different groups;and use Western blot to measure p65,p-p65,IκBα,p-IKBa in the corresponding groups on tumor tissues.Results:In the cell experiment,compared with the blank control group,Guzheng Xiaoai Decoction can inhibit human colon LOVO cells,HT-29 cells,and HCT-116 cells,and make the cell morphology irregular in a dose-dependent manner.Changes and significantly affected the proliferation of human colon cancer cells.In terms of cell morphology,the optical inverted microscope showed that LOVO cells,HT-29 cells,and HCT-116 cells in the blank control group had clear appearance and normal growth morphology.In the Guzheng Xiaoai Decoction group,the 3 cell morphologies showed irregular morphological changes,and the number and growth of the cells showed a tendency of attenuation as the concentration of the drug-containing serum increased.The CCK-8 method was used to detect the OD values of LOVO cells,HT-29 cells,and HCT-116 cells in the serum of Guzheng Xiaoai Decoction after 24h,48h and 72h culture(P<0.05),and HT-29 cells are the most sensitive to the concentration gradient of serum containing Guzheng Xiaoai decoction.Follow-up with HT-29 cells as the research object,with the increase of the serum concentration of Guzheng Xiaoai Decoction,the proliferation and migration ability of HT-29 cells are also decreasing.Western blot method was used to detect the protein levels of p65,p-p65,IκBα,and p-IKBa in HT-29 cells.Under the intervention of the medicated serum of Guzheng Xiaoai Decoction,compared with the blank control group,the protein expression of p-IKBa decreased significantly(P<0.05),and there were statistical differences between different doses and concentrations(P<0.05).This dose-dependent relationship is also reflected in the expression of p-p65 protein.After low,medium and high doses of contained Guzheng Xiaoai Decoction groups acted on HT-29 cells,the expression of p-p65 protein decreased compared to the blank control group.And there are significant differences between the groups(P<0.05).Nude mice use HT-29 cells to construct a tumor-bearing nude mouse model.In the experiment of tumor-bearing nude mice,Guzheng Xiaoai Decoction can improve the living quality of nude mice to a certain extent;the pathological results of the tumor body HE staining show that the Guzheng Xiaoai Decoction administration group can promote tumors cell apoptosis and necrosis more than the model group;tumor immunohistochemical results believe that Guzheng Xiaoai Decoction can reduce Bcl-2 protein and increase the level of Bax protein;p65 and p-p65 in tumor tissues detected by Western blot method.The protein content of IKBα,IκBα and p-IKBa are consistent with the results of cell experiments,and inhibit the NF-κB signal pathway conduction.It shows that Guzheng Xiaoai Decoction can also promote the apoptosis of colorectal cancer cells in the body and limit the expansion and proliferation of cancer tissues.Conclusions:Guzheng Xiaoai Decoction related to the inhibition of NF-κB signal pathway has a good effect of inhibiting the colorectal cancer.