Compound A Mortar Concurrent Chemotherapy Attenuated The Synergistic Effect And Its Mechanism Of Experimental Research

Comprehensive treatment of malignant tumors has become a model in anti-tumor therapy. With the progress on the study of traditional Chinese medicine in the treatment of cancer and the updates annually in research methods, various formulations of Chinese medicine integrated in the treatment of malignant tumors have got much attention from scholars in our country and abroad. There are lots of clinical and experimental research in the treatment of malignant tumors with Chinese medicine in recent years, which suggest the unique advantages and certain efficacy of Chinese medicine in alleviating clinical symptoms, prolonging survival time with tumor, improving the quality of life, toxicity reducing and efficacy enhancing of chemotherapy. Many Chinese patent drugs have been listed, such as Xihuang pills, Fufang Banmao capsule, Shenlian capsule, Pingxiao capsule, Jinlong capsule, Shenyi capsule, Weimaining, Zilongjin, Zijinlong capsule, Compound Kushen injection, Xiaoaiping injection, Aidi injection, Kang ai injection, Kanglaite injection and Chemoradiotherapy adjuvants such as Zhenqifuzheng granule, Jianpiyishen Granules, Bailing capsule, Yizhong shengxue capsule, and Shenqi Fuzheng Injection. Besides that, the study in the anti-tumor mechanism of traditional Chinese medicine has also made lots of achievements, which mainly centered on the tumor cell proliferation, differentiation, apoptosis, oncogenes and tumor suppressor genes, and immune regulation. However, the experimental study on toxicity reducing has not deep enough, and especially the research on toxicity decreasing but not effect reducing with both anti-tumor effects of traditional Chinese medicine is still a little. So it is necessary to actively carry out basic experimental research in this field.1 ObjectiveIn this study, lewis lung cancer xenograft model were made, and treated with Compound Ejiao slurry (CEJS) and chemotherapy drug cyclophosphamide (CTX). Explore the influence of CEJS on the toxic and side effects of chemotherapy agents, from the peripheral blood and immune functions associated with the toxicity of chemotherapy, especially for the protection of blood, to the tumor inhibition rate and anti-tumor mechanism. Observe the anti-tumor effect of the drug and further seek the possible anti-tumor mechanism for CEJS in order to provide new experimental basis and scientific guidance for the clinical application of CEJS.2 MethodsLewis lung cancer xenograft model were made by armpit injection using lewis cells of lung cancer. C57BL/6 mice were randomly divided into model group, CTX group, CEJS group(medium dosage), CEJS high, medium, low-dosage plus CTX groups(combination group 1,2,3) according to latin table method. Thirteen days were observed. The measurement of blood routine were carried out on day 7th, 13th. The weight of mice and tumor volume were measured every four days before the experiment and after. Drew the curve of weight changes and tumor growth. The volume were measured with a vernier caliper. Calculated thymus and spleen index, and tested lymphocyte proliferation function with MTT method after the experiment. Removed the tumor and calculated the inhibitory rate. The expression of vascular endothelial growth factor VEGF, apoptosis protein Bcl-2/Bax, CyclinD1, and adhesion molecules CD44 were detected by immuno-histochemistry. Compared the survival rate of mice in each group.3 Results3.1 Study on the attenuation effect of CEJS on chemotherapy drug3.1.1 Effects of CEJS on hematological toxicity of chemotherapy drugOn the 7th day after treatment, the CTX group declined more obviously, compared the numerical value of WBC with other groups (p<0.05～0.01).The WBC amount of each combination group rised significantly than CTX group (p<0.05～0.01). There were no significant difference between the experimental groups and model group in the number of RBC, HGB, and PLT (p>0.05). On the 13th day after treatment, the WBC amount of CTX and every combination group increased more rapidly compared with the model group (p<0.01). Compared the numerical value of RBC and HGB with the model group, the CTX and combination group 3 were lower significantly (p<0.01). The numerical value of RBC and HGB increased in combination group 2 compared with CTX group (p<0.05). Each group had no significant difference in platelet count (p>0.05).3.1.2 CEJS on the regulation of immune functionThe spleen index in the CEJS group and combination groups increased significantly compared with the control group (p<0.01). The spleen index in combination group 1 and 2 raised significantly compared with the CTX group (p<0.05).The thymus gland index in CEJS increased significantly (p<0.01). The proliferation function of spleen cells in CEJS and combination group 2 were stronger than the control and CTX group (p<0.05).3.2 Study on the synergy effect of CEJS and the mechanism of anti-tumor effectCompared with model group, administration could reduce the tumor weight in each group (p<0.05). CEJS had an inhibition rate of 50%. The combination administration groups had no significant difference between CTX group (p>0.05). Inhibition rate after combination therapy were increased by 9.78%(combination group 1),7.79%(combination group 2). There was a trend of inhibition rates of CTX increasing by CEJS, with the same results from tumor volume curve.The expression level of VEGF could be significantly down-regulated in CTX or combination group 1 and 2, compared with the control group (p<0.05). CEJS group had no significant difference with the control (p>0.05). The expression in combination group 1 and 2 were not significantly different with the CTX group (p>0.05). The expression level of Bcl-2 could be significantly down-regulated and the level of bax could be significantly up-regulated in each experimental group, compared with the control group (p<0.01～0.05). Compared with the CTX group, the expression of Bcl-2 were obviously lower in combination group 1 (p<0.05). The expression of bax were significantly higher in combination group 2, compared with the CTX group (p<0.01). CEJS and combination group 1 resulted in more significant suppression of the expression of CyclinDl when compared with the model group (p<0.05). The expression of CD44 in CTX group, CEJS group, combination groupl,2 were lower than the model group (p<0.05).4 Conclusions4.1 Successfully prepared lewis lung cancer xenograft model in mice. White cell amounts in CTX group significantly reduced compared with other groups on the 7th day after administration. RBC and HGB amounts in CTX group significantly reduced compared with other groups on the 13th after administration. CEJS could effectively recover WBC, RBC, HGB count and promote the bone marrow hematopoietic function.4.2 CEJS could increase the spleen and thymus index of tumor-bearing mice, and promote proliferation of spleen lymphocytes, suggesting that it may have a moderating effect on immunity.4.3 CEJS could not improve the effects of chemotherapy obviously, but it itself had some anti-tumor effect, which may be associated with down-regulation of apoptosis-related protein Bcl-2, Bax and down-regulation of cyclin protein CyclinD1, and adhesion CD44 molecules.Summary:CEJS had toxic-reducing effects of chemotherapy, reflected in two aspects:①Protected the decreased blood caused by CTX.②Enhanced immune function. Reduced the damage of bone marrow hematopoietic function, also played an anti-tumor effect. CEJS did not affect the efficacy of chemotherapy drugs only, but also had a synergistic trend to a certain extent.