| Objective:to study the effect of Shenling Baizhu Powder modified compound on VEGF and m TOR pathway related proteins in mice with bone cancer pain,and to explore the possible mechanism of Lingbaizhu Powder modified compound in the treatment of bone cancer pain.Methods:in 58 male BALB/c mice,9 mice were randomly selected as normal group,and the other 49 mice were used to establish the model of bone metastasis and pain of lung cancer.4 mice died during the modeling process,which were not included in the grouping.The other 45 model mice were randomly divided into two groups.Divided into model groups,Shenling Atractylodes macrocephala powder high,medium and low dose group,rapamycin group,a total of 5 groups,9 in each group,normal group,model group normal feeding;The mice in the high,middle and low dose groups of Shenling Baizhu Powder were given intragastric administration of 0.4ml daily for 7 days at the 6th week after modeling,and those in the rapamycin group were given intraperitoneally according to the dose of 4mg/kg once a day for 7days at the 6th week after modeling.The model was evaluated by behavioral changes,HE staining and pain behavior detection in mice,and the therapeutic effect of Shenling Baizhu Powder modified compound on bone cancer pain mice was evaluated.Tunel staining was used to observe the apoptosis of tumor cells in each group.The expression of VEGF and m TOR protein in tibia tissue of mice in each group was detected by immunohistochemistry and Western blot.Results:1.According to the behavioral changes of each group in the first week after the establishment of the model,the comparison of HE staining between the normal group and the model group at the 2nd week after the establishment of the model,and the comparison of the pain behavior of the normal group and the model after the establishment of the model,suggesting that the model was established successfully.2.The pain behavior of mice in each group showed that the pain threshold of mice in Shenling Baizhu powder group and rapamycin group was significantly higher than that in model group(P < 0.01).The pain behavior of mice in each group showed that the pain threshold of mice in Shenling Baizhu powder group and rapamycin group was significantly higher than that in model group(P < 0.01).Compared with the rapamycin group,the middle and low dose groups,There was no significant difference(P > 0.05).3.HE staining showed that the number of tumor cells in tibia bone marrow cavity in Shenling Baizhu powder group was lower than that in model group,and the number of bone marrow cells in tibia bone marrow cavity in dose group was about uniform,and the number of tumor cells in tibia bone marrow cavity in Shenling Baizhu powder group was lower than that in model group.There were a small number of tumor cells and slight changes in bone structure.The number of tumor cells in the bone marrow cavity of mice in rapamycin group decreased,but at the same time,there was obvious bleeding.4.The results of immunohistochemistry showed that compared with the model group,the positive rates of VEGF and m TOR protein in the high,middle and low dose groups and rapamycin group were significantly lower than those in the model group(P < 0 05).Compared with rapamycin group,the positive rate of VEGF and m TOR protein in middle dose group was significantly lower than that in rapamycin group(P < 0.01).5.The results of Tunel staining showed that compared with the model group,the apoptosis of tumor cells in Shenling Baizhu powder group,middle dose group and rapamycin group was significantly higher than that in the model group(P < 0 01).The results of DNA staining showed that the apoptosis of tumor cells in Shenling Baizhu powder group and rapamycin group were significantly higher than those in model group.There was no significant difference between rapamycin group and middle dose group(P > 0.05).6.Western blot test showed that compared with the model group,the expression of VEGF and m TOR protein in high,middle and low dose groups and rapamycin group were significantly lower than those in the model group(P < 0 01).The results showed that compared with the model group,the protein expression of VEGF and m TOR in Shenling Baizhu powder group was significantly lower than that in the model group(P < 0 01).Conclusion:1.The mouse model of bone metastasis and pain of lung cancer was successfully established.2.Shenling Baizhu Powder modified compound can relieve the pain of bone cancer pain mice and promote the apoptosis of tumor cells.3.Shenling Baizhu Powder modified compound can down-regulate the expression of VEGF and m TOR protein in tibia tissue of model mice.4.Shenling Baizhu Powder modified compound may play a therapeutic role in mice with bone cancer pain by inhibiting the expression of VEGF and m TOR,thus affecting tumor angiogenesis and blocking the conduction of bone cancer pain signal. |
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