| BackgroundBy observing the tumor volume,tumor weight,survival time,immunophenotype of CD3~-/NKp46~+NK cells,CD4~+,CD8~+T cells in tumor environment and the tumor microenvironment and CD3~-/NKp46~+NK cell activity in tumor microenvironment of Lewis lung cancer tumor-bearing mice,to study the effect of grain-sized moxibustion(g Moxi)on tumor suppression and immunity in Lewis lung cancer tumor-bearing mice,and to explore the effect and mechanism of grain-sized moxibustion on NSCLC.Methods1.Evaluation of tumor regulation effect of grain-sized moxibustion(g Moxi)with different amounts on Lewis lung cancer tumor-bearing mice:Lewis lung cancer(LLC)xenograft tumor model was established,and grain-sized moxibustion(g Moxi)was performed at Zusanli(ST36)acupoint.To observe the regulation effect of grain-sized moxibustion(g Moxi)with different amounts on Lewis lung cancer tumor-bearing mice and select the appropriate amount of moxibustion.2.Effect of grain-sized moxibustion(g Moxi)on the survival time of Lewis lung cancer tumor-bearing mice:Lewis lung cancer(LLC)xenograft tumor model was established,and grain-sized moxibustion(g Moxi)was performed at Zusanli(ST36)acupoint.By measuring the tumor volume of tumor-bearing mice,observe the effect ofgrain-sized moxibustion(g Moxi)on the survival of tumor-bearing mice.3.Effect of grain-sized moxibustion(g Moxi)on immunophenotype of CD3~-/NKp46~+NK cells,CD4~+,CD8~+T cells in tumor environment and the tumor microenvironment and CD3~-/NKp46~+NK cells activity in tumor microenvironment of Lewis lung cancer tumor-bearing mice:Lewis lung cancer(LLC)xeograft tumor model was established,and grain-sized moxibustion(g Moxi)was performed at Zusanli(ST36)acupoint.Flow cytometry was used to detect the influence of immunophenotype of CD3~-/NKp46~+NK cells,CD4~+,CD8~+T cells in tumor environment and the tumor microenvironment of Lewis lung cancer tumor-bearing mice.Annexin V/PI double staining was used to detect the changes in the activity of CD3~-/NKp46~+NKs cell activity in tumor microenvironment of Lewis lung cancer tumor-bearing mice,and further immunofluorescence was used to detect the infiltration of CD3-/NKp46+NK cells in tumors of tumor-bearing mice.4.The grain-sized moxibustion depended on NK cells inhibiting the tumor growth innon-small cell lung cancer:NK cells antibody PK136 was used.Lewis lung cancer(LLC)xenograft tumor model was established,and grain-sized moxibustion(g Moxi)was performed at Zusanli(ST36)acupoint.Flow cytometry was used to detect the influence of immunophenotype of CD3~-/NKp46~+NK cells,CD4~+,CD8~+T cells in tumor environment and the tumor microenvironment of Lewis lung cancer tumor-bearing mice.Annexin V/PI double staining was used to detect the changes in the activity of CD3~-/NKp46~+NK cells in the internal environment of tumor-bearing mice,and further immunofluorescence was used to detect the infiltration of CD3~-/NKp46~+NK cells in tumors of tumor-bearing mice.5.Tumor RNA sequencing(RNA-seq)analysis and verification of the tumor inhibition effect of grain-sized moxibustion(g Moxi)on Lewis lung cancer tumor-bearing mice:Through RNA sequencing(RNA-seq)of tumor-bearing mice,the possible mechanism of grain-sized moxibustion(g Moxi)delaying tumor growth of NSCLC was screened out and subsequently verified.Results1.Evaluation of tumor regulation effect of grain-sized moxibustion(g Moxi)with different amounts on Lewis lung cancer tumor-bearing mice:compared with the model group,g Moxi-3 and g Moxi-7 groups significantly inhibited LLC tumor growth,and the difference was statistically significant(All P<0.05).There was no significant difference in inhibitory effects betweeng Moxi-3 group and g Moxi-7 group(P>0.05),Therefore,the subsequent experiments used g Moxi-3 on bilateral posterior Zusanli(ST36) acupoint,every other day.2.Effect of grain-sized moxibustion(g Moxi)on the survival time of Lewis lung cancer tumor-bearing mice:compared with the model group,the g Moxi group significantly prolonged the survival time of tumor-bearing mice(P<0.05).3.Effect of grain-sized moxibustion(g Moxi)on immunophenotype of CD3~-/NKp46~+NK cells,CD4~+,CD8~+T cells in tumor environment and the tumor microenvironment and CD3~-/NKp46~+NK cell activity in tumor microenvironment of Lewis lung cancer tumor-bearing mice:compared with the model group,the g Moxi group significantly increased the proportion of CD3~-/NKP46~+NK cells in the internal environment and tumor microenvironment and the activation of CD3~-/NKP46~+NK cells in the internal environment of Lewis lung cancer tumor-bearing mice(all P<0.05).Compared with the model group,the g Moxi group increased the infiltration of CD3~-/NKP46~+NK cells in the tumor of LLC tumor-bearing mice(P<0.05),and the proportion of CD3~+CD4~+T cells and CD3~+CD8~+T cells in the internal environment and tumor microenvironment of Lewis lung cancer tumor-bearing mice had no significant difference(all P>0.05).4.The grain-sized moxibustion depended on NK cell inhibiting the tumor growth in non-small cell lung cancer:compared with the model group,the PK136 group reduced the proportion of CD3~-/NKP46~+NK cells in the internal environment and tumor microenvironment and the activation of CD3~-/NKP46~+NK cells in the internal environment of Lewis lung cancer tumor-bearing mice(all P<0.05).Compared with the model group,the proportion of CD3~-/NKP46~+NK cells in the internal environment and tumor microenvironment and the activation of CD3~-/NKP46~+NK cells in the internal environment of Lewis lung cancer tumor-bearing mice in the PK136+g Moxi group were significantly reduced(all P<0.05).Compared with the model group,PK136 group and PK136+g Moxi group showed no significant inhibitory effect on tumor growth in Lewis lung cancer tumor-bearing mice(all P>0.05).There was no significant difference in the proportion of CD3~+CD4~+T cells and CD3~+CD8~+T cells in the internal environment and tumor microenvironment of Lewis lung cancer mice in each group(all P>0.05).5.Tumor RNA sequencing(RNA-seq)analysis and verification of the tumor inhibition effect of grain-sized moxibustion(g Moxi)on Lewis lung cancer tumor-bearing mice:A total of 689 genes were differentially expressed between the wheat moxibustion group and the model group,among which 346 genes were significantly down-regulated and 343 genes were significantly up-regulated.The number of differentially expressed genes was statistically analyzed and the top 30 signaling pathways were selected according to the enrichment degree.Most of them are related to neuroendocrine signaling pathways,among which the adrenaline signaling pathway has attracted the attention of this study.6.Effect of adrenaline signaling pathway on grain-sized moxibustion to enhance anti-tumor immunity of NK cells:compared with the model group,propranolol group significantly inhibited tumor growth of Lewis lung cancer tumor-bearing mice,and promoted the proportion of CD3~-/NKP46~+NK cells in the internal environment and tumor microenvironment and the activation of CD3~-/NKP46~+NK cells in the internal environment of Lewis lung cancer tumor-bearing mice(all P<0.05);Compared with propranolol group,the propranolol+g Moxi group did not inhibit tumor growth but increased the proportion of CD3~-/NKP46~+NK cells in the internal environment and tumor microenvironment and the activation of CD3~-/NKP46~+NK cells in the internal environment of Lewis lung cancer tumor-bearing mice(all P>0.05).Compared with the model group,the epinephrine group inhibited the proportion of CD3~-/NKP46~+NK cells in the internal environment and tumor microenvironment and the activation of CD3~-/NKP46~+NK cells in the internal environment of Lewis lung cancer tumor- bearing mice(all P<0.05).Compared with epinephrine group,epinephrine+g Moxi group significantly inhibited tumor growth in Lewis lung cancer tumor-bearing mice,and increased the proportion of CD3~-/NKP46~+NK cells in the internal environment and tumor microenvironment and the activation of CD3~-/NKP46~+NK cells in the internal environment of Lewis lung cancer tumor-bearing mice(all P<0.05).There was no significant difference in the proportion of CD3~+CD4~+T cells and CD3~+CD8~+T cells in the internal environment and tumor microenvironment of Lewis lung cancer mice in each group(all P>0.05).Conclusion1.Grain-sized moxibustion can inhibit the growth of non-small cell lung cancer tumor,can prolong the survival time of Lewis lung cancer-bearing mice.2.Grain-sized moxibustion increased the proportion of NK cells in the internal environment and tumor microenvironment and activation of NK cells in the internal environment of Lewis lung cancer tumor-bearing mice,and enhanced the infiltration of NK cells in tumors.3.Grain-sized moxibustion inhibits tumor growth in Lewis lung cancer tumor-bearing mice,and tumor regression by Grain-sized moxibustion depended on NK cells and Grain-sized moxibustion inhibits adrenaline signaling pathway and then promotes the activation of NK cells to anti-tumor. |
PDF Full Text Request