| Background:Retro-zuojin Pill was inspired by Zuojin Pill,which was composed of 6?1 ratio of fructus evodiae rhizoma coptidis.Modern pharmacology research shows that the anti-inflammatory anti-bacterial,anti-ulcer,analgesic and so on antiphlogistic anti-bacterial effects.Anti-tumor studies have shown that both Berberine and Evodiamine can block the activation of cancer-promoting substances to the potential cancerous cells and thus play an anti-tumor role.Oxaliplatin is the most common and effective chemotherapy drug for colon tumors.However,in the clinical application of L-OHP,it was found that it was easy to cause gastrointestinal toxicity and drug resistance,which affected the postoperative recovery of colon cancer patients.In recent years,a large number of clinical studies on the treatment of colon cancer with the combination of traditional Chinese and western medicine have shown that,after the operation of colon cancer patients,the treatment of colon cancer with targeted drugs of traditional Chinese medicine combined with chemotherapy can not only effectively alleviate the toxic and side effects of digestive tract caused by the process of radiotherapy and chemotherapy,but also can improve the immunity of the body to fight the recurrence and metastasis.Retro-zuojin pills with different concentrations combined with L-OHP were designed to treat colon cancer in nude mice for experimental study,in order to explore the best and effective combination of traditional Chinese and western medicine treatment,and to provide some new ideas for the clinical treatment of colon cancer patients.Objective:The purpose of this study was to investigate the effects of Retro-zuojin pill combined with L-OHP on the expression of IL-10,IL-2,and NF-?B p65 in tumor-bearing nude mice with human colon cancer SW480 cells,and to explore the mechanism of action.Methods:Colon cancer SW480 cells in logarithmic growth stage were collected,and their concentration was adjusted to become 5.5×10~6 cell suspension.0.2ml/dose was injected into the right underarm of 5 nude mice for modeling.Tumor growth was observed in nude mice.With trypan blue staining,tumor tissues with better activity were selected,cleaned with medium,cut into 2 mm~3 or so,and inoculated into 60nude mice under the right armpit.After the tumor grew to about 100 mm~3,the tumor was randomly divided into 6 groups,10 in each group,namely,the blank control group(the single tumor model group),the positive control group(the L-OHP group),the low dose Retro-zuojin pill group,the high dose Retro-zuojin pill group,the low dose Retro-zuojin pill+L-OHP group,and the high dose Retro-zuojin pill+L-OHP group.Then,the body weight and tumor volume of nude mice were measured at regular intervals after 4 weeks of continuous drug administration.On the next day after drug withdrawal,the body weight and tumor volume of nude mice were measured by eyeball and blood collection ELISA kit.Results:1.The mean tumor volume in each group was 1208±41.11mm~3;831.3±38.82mm~3;998.5±18.54mm~3;906.7±4.989mm~3;489±16.33mm~3;307±15mm~3.Compared with the blank control group,the tumor volume of nude mice in the L-OHP group,the low-dose group of Retro-zuojin pill,the high-dose group of Retro-zuojin pill,the low-dose group of Retro-zuojin pill+L-OHP group and the high-dose group of Retro-zuojin pill+L-OHP group were significantly reduced(p<0.05).Compared with the L-OHP group,the tumor volume of nude mice in the low-dose group and the high-dose group of the Retro-zuojin pill was significantly larger and statistically insignificant(p>0.05),while the tumor volume of nude mice in the low-dose+L-OHP group and the high-dose+L-OHP group was significantly reduced(p<0.05).Compared with the low-dose group,the tumor volume of nude mice in the low-dose+L-OHP group was significantly reduced(p<0.05).The tumor volume of nude mice in the group of L-OHP was significantly reduced(p<0.05).Compared with the blank control group,the tumor inhibition rates of the L-OHP group were 31.2%,17.4%,25.0%,59.5%and 74.5%,respectively,in the L-OHP group,the low-dose group,the high-dose group,the low-dose group,the low-dose group and the high-dose group,respectively(p<0.05).2.ELISA results showed that the mean value of IL-10 in each group was68.95±4.691,35.81±2.909,36.53±3.259,38.69±5.384,53.55±2.956,66.25±5.489.The mean values of IL-2 in each group were 27.78±3.149,56.59±5.064,48.6±2.837,43.8±1.978,34.58±2.584,and 29.94±1.544.Compared with blank control group,L-OHP,the Retro-zuojin pill to low dose group,the levels of IL-10 Retro-zuojin pill dose group were falling,IL-2 levels,the difference was statistically significant(p<0.05),but the Retro-zuojin pill to low dose+L-OHP group,the Retro-zuojin pill+L-OHP group of IL-10 levels drop,IL-2 levels,differences statistically insignificant(p>0.05);Compared with the L-OHP group,the level of IL-10 in the low-dose group and the high-dose group was increased and the level of ILl-2 was decreased,but the difference was statistically insignificant(p>0.05).However,the IL-10 level increased and the IL-2 level decreased in the low dose+L-OHP group and the high dose+L-OHP group respectively(p<0.05),and the difference between the two groups was proportional to the concentration of Retro-zuojin pill.3.compared with the blank control group,the IL-10 level of the positive control group decreased and the IL-2 level increased.Compared with the positive control group,the level of IL-10 increased and the level of IL-2 decreased in the high and low dose group.Compared with the positive control group,the level of IL-10increased and the level of IL-2 decreased in the low dose Retro-zuojin pill combined with L-OHP group.The differences between groups were statistically significant(p<0.05).Compared with the positive control group,the level of IL-10 increased and the level of IL-2 decreased in the L-OHP group combined with the high dose of Retro-zuojin pill(p<0.0001).4.Immunohistochemical staining results showed that compared with the blank control group,the expression levels of NF-?B p65 in the L-OHP group,the low-dose group of Retro-zuojin pill,the high-dose group of Retro-zuojin pill,the low-dose group of Retro-zuojin pill+L-OHP group and the high-dose group of Retro-zuojin pilll+L-OHP group decreased significantly.Compared with the positive control group,the expression levels of NF-?B p65 were increased in the low-dose group and the high-dose group,but the expression levels of NF-?B p65 were significantly decreased in the low-dose group and the high-dose group.Conclusions:Retro-zuojin pill combined with L-OHP has obvious anti-tumor effect on the tumor-bearing nude mice with colon cancer SW480 cells.The mechanism may be to inhibit the production of pro-inflammatory factor IL-2 and promote the secretion of anti-inflammatory factor IL-10 by blocking the activation of NF-?B p65. |
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