| Objective:Liver is the most common distant metastasis organ of colorectal cancer.Tumor metastasis is closely related to immunity.In this experiment,we explored the effect of moxibustion on immune cells in nude mice model of liver metastasis of colonrectal cancer,and explored the mechanism of moxibustion in reducing liver metastasis based on inherent immunity.Methods:This experiment is divided into four parts:(1)The spleen injection fluorescent labeled HCT-116 colorectal cells for liver metastasis model was established.The tumor in the spleen and the liver was observed under a fluorescence detector twice a week,and typical fluorescent photographs were taken.The nude mice were sacrificed 7 weeks after the surgery,and the spleen and liver tumors were recorded,then it was evaluated whether the modeling surgery was successful which was prepared for the subsequent research;(2)In the second part,The nude mice were randomly divided into model group,moxibustion treatment group,moxibustion prevention+treatment group and preventive moxibustion group.There were 15 mice in each group.Moxibustion at Ganshu(BL18)and Zusanli(ST36)points on both sides were selected to observe the mental state of nude mice,the changes of body weight,survival status,survival rate,tumor formation rate,liver metastasis rate,tumor weight and tumor volume of nude mice in each group;(3)Flow cytometry was used to detect the ratio and expression of NK cells,macrophages and neutrophils in peripheral blood,liver and spleen of nude mice in each group;(4)Random numbers were generated using SAS software,and Balb/c nude mice were divided into control group and moxibustion group,with 8 rats in each.Moxibustion at Ganshu and Zusanli points on both sides was treated in the treatment group.Flow cytometry was used to detect the content and the rate of activated NK cells in peripheral blood,liver and spleen of nude mice.The expression of NK cell receptor(NKp46)was detected by immunohistochemistry.Results:(1)In the first part of the experiment,the model of liver metastasis of colon cancer in nude mice was successfully established.Tumor tissues were found in both spleen and liver organs.The metastasis rate was 100%and the mortality rate was 33.3%,which was similar to previous literature reports.(2)In the second part of the experiment,we found that:the model group had the lowest animal survival rate of 66.7%,and the moxibustion treatment group had the highest survival rate of 86.7%.The death of cachexia in the moxibustion group and the preventive moxibustion group occurred later.There was no significant differences in survival rate,cachexia mortality,and the time of occurrence of cachexia between the groups(P>0.05);Compared with the model group,the prevention of moxibustion was not conducive to the normal increase of body weight in nude mice,but showed inhibition of body weight loss in nude mice with metastasis after 4 weeks from the model establishment,and the difference was statistically significant(P<0.05);Compared with the model group and the preventive moxibustion group,the weight and the size of spleen tumor,,in nude mice in the moxibustion treatment group and the moxibustion prevention+treatment group were smaller,and the difference was statistically significant(P<0.05);Compared with the model group,the liver metastasis rate of the three moxibustion groups decreased significantly,and the difference was statistically significant(P<0.05).(3)In the third part,the content of innate immune cells(in peripheral blood,liver and spleen)of nude mice with colon cancer liver metastasis was detected by flow cytometry and the results were as follows:,Compared with the model group,the rate of activated NK cells(CD 107a ratio)in peripheral blood among the other three moxibustion groups did not change,and the difference was not statistically significant(P>0.05);In the liver,compared with the model group,the rate of activated NK cells in the moxibustion treatment group was significantly increased(P<0.05);in the spleen,compared with the model group,the rate of activated NK cells in each moxibustion group did not change,and the difference was not statistical significance(P>0.05);In terms of macrophages in spleen,compared with the model group,the proportion of macrophages in the moxibustion treatment group and the moxibustion prevention+treatment group was higher and had statistically significant difference(P<0.05);There was no significant difference in macrophage content between the groups in the liver or in the peripheral blood(P>0.05);In terms of neutrophils,there was no significant difference in the proportion of neutrophils in peripheral blood,liver and spleen between the groups(P>0.05).(4)The fourth part of the research results:In peripheral blood,there was no significant change in the number of NK cells(CD49b and CD335 ratio)and NK cell activation rate between the control group and the moxibustion group(P>0.05);In the liver tissue,compared with the control group,the NKcell number and the rate of activated NK cells in the moxibustion group increased(P<0.05);In spleen tissues,there was no significant change in the number of NK cells or the rate of activated NK cells between the groups(P>0.05).At the same time,the immunohistochemical results were similar,the IOD values of each group did not change significantly(P>0.05).Conclusion:Moxibustion can activate the body’s innate immune system especially by increasing the activation rate of NK cells in liver tissue and the content of macrophages in spleen tissues,effectively inhibit the growth of the tumor implanted in the spleen,rapidly kill the tumor cells of the colon cancer metastasized to the liver;Besides,the treatment of moxibustion has a certain universality for the up-regulation of the rate of activated NK cells in the liver,which embodies the idea of "Strengthening the body resistance" of TCM and can provide experimental basis for moxibustion treatment of other neoplastic or immunodeficiency disease. |
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