| ObjectiveIn this study,the effect of sshenqi anti-tumor decoction on the immune function of H22liver cancer tumor-bearing mice and the expression of Bcl-2,Bax,Caspase-3 and CytC genes and proteins in tumor tissues was studied to explore the inhibitory effect of shenqi anti-tumor decoctionon liver cancer.Methods70 Kunming mice were purchased,weighing(20±2)g,half male and half female,10 of them were used for inoculation of ascites,and the remaining 60 were inoculated with H22liver cancer ascites cells.After modeling,the mice were randomly divided into six groups,10in each group,half male and half female.The groups were model group,cyclophosphamide group,Shenqi anti-tumor decoction high-dose group(Chinese medicine high-dose group),Shenqi anti-tumor decoction middle-dose group(Chinese medicine middle-dose group),Shenqianti-tumor decoction low-dose(Chinese medicine low-dose group),Shenqi anti-tumor decoction middle-dose combined with cyclophosphamide group(combination group).Drug intervention was given on the second day after grouping.The high,middle and low dose groups of Shenqi anti-tumor decoction were given 193.75g/kg,96.86g/kg,19.38g/kg respectively.The combination group and cyclophosphamide group were given cyclophosphamide 30mg/kg daily.once a day by intragastric administration for 15 days.At the end of the 15th day,the mice were fasted for 24 hours,and 24 hours later,the eyeballs of the mice were taken for blood collection,and then the tumor tissues and spleen tissues were removed.Observed and analyzed the results:(1)general life state;(2)spleen index,tumor weight and tumor inhibition rate;(3)tumor histopathological changes;(4)changes in mouse blood routine;(5)mouse peripheral CD3~+,CD4~+,CD8~+levels in blood;(6)Bcl-2,Bax,Caspase-3,CytC related gene expression in tumor tissues;(7)Bcl-2,Bax,Caspase-3,CytC in tumor tissues Related protein expression.Results1.The vital signs and general activity status of mice in each group were different.The results of weight change of mice in each group during drug administration showed that:compared with the model group,the weight of mice in each treatment group decreased,the weight loss was most significant in the high-dose group and the combination group of traditional Chinese medicine,and the weight loss was next in the low-dose group,medium dose group and cyclophosphamide group,the difference was statistically significant(P<0.05).2.Spleen index results showed that,compared with the model group,the spleen index of all treatment groups was increased,significantly increased in the combination group,the high-dose group of traditional Chinese medicine and the cyclophosphamide group,and slightly increased in the medium and low-dose group of traditional Chinese medicine,with statistically significant difference(P<0.05).Compared with the cyclophosphamide group,the spleen index of the high-dose group and the combined group were significantly increased,and the difference was statistically significant(P<0.05).3.The results showed that,compared with the cyclophosphamide group,the inhibitory rate of both the high-dose group and the combined group increased,with statistically significant differences(P<0.05).4.Blood routine results showed that:compared with the model group,the white blood cells in each treatment group decreased,and the cyclophosphamide group significantly decreased,with statistically significant difference(P<0.05).Compared with the cyclophosphamide group,the white blood cells of each treatment group and the combined treatment group were increased,and the difference was statistically significant(P<0.05).Compared with the model group,erythrocytes in each treatment group were increased,and the difference was statistically significant(P<0.05).Compared with the cyclophosphamide group,erythrocytes in the high-dose group,the medium-dose group and the combined group were significantly increased,with statistically significant differences(P<0.05).Compared with the model group,platelets in each treatment group decreased,and the difference was statistically significant(P<0.05).Compared with the cyclophosphamide group,platelets in the high,medium and low dose groups and the combination group were significantly increased,with statistically significant differences(P<0.05).5.Flow cytometry results showed that,compared with the model group,the expression of CD3~+and CD4~+T cells in each treatment group was increased,with statistically significant differences(P<0.05).Compared with cyclophosphamide group,the expression of CD3~+T and CD4~+T cells in the high-dose group,the medium-dose group and the combined group were significantly increased,with statistically significant differences(P<0.05).Compared with the model group,the expression of CD8~+T cells in each treatment group was decreased,and the difference was statistically significant(P<0.05).Compared with the cyclophosphamide group,the expression of CD8~+T cells in the high-dose group,the medium-dose group and the combined group were significantly decreased,and the difference was statistically significant(P<0.05).6.Tumor tissue pathology results:compared with model group,the Chinese medicine treatment group,cyclophosphamide group and combination group in the cells of the necrotic cells on the relatively loose,dyed lighter,smaller,there are different amount of dead cells,in traditional Chinese medicine(TCM)in high dose group and combination group most pronounced dead cells,low dose group and cyclophosphamide group is relatively small.7.Gene and protein expression result shows:compared with model way,the treatment group the Bcl-2 mRNA and protein expression decreased,high dose group,a significant reduction in the cyclophosphamide group and combination group,the difference is statistically significant(P<0.05),compared with cyclophosphamide group,high dose group and combination group of Bcl-2 mRNA and protein expression were markedly reduced,statistically significant difference(P<0.05);Compared with model group,the treatment group in the tumor tissue Bax,Caspase-3,CytC mRNA and protein expression of rise,high dose group,a significant rise in cyclophosphamide group and combination group,difference has statistical significance(P<0.05),compared with cyclophosphamide group,high dose group and combination group of Bax,Caspase-3,CytC mRNA and protein expression were significantly elevated,statistically significant difference(P<0.05).ConclusionShenqi anti-tumor decoction can induce apoptosis of H22 hepatoma cells in tumor-bearing mice through mitochondrial apoptosis pathway.Shenqi anti-tumor decoctioncombined with cyclophosphamide can effectively improve the body’s immunity,promote cancer cell apoptosis,and inhibit tumor recurrence and metastasis.Delay the growth rate. |
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