| Objective The syndrome of "liver-stagnation and spleen-deficiency (LSSD)" had become one of the common clinical syndromes in Traditional Chinese Medicine (TCM), and it had been exerted widely in the differentiation and typing of tumor diagnosis. The reasons that cause LSSD were negative psychological factors and irregular diet .And these also play important roles in the development of tumors. Therefore, The mouse and rat models of "integrated LSSD and tumor" were set up in our research. The T cellular immune function and Th1/Th2 balance in lump- bearing mice were observed. The cell cycles of ascitic fluid tumor cells in mice were detected. And the gene expression of ornithine decarboxylase (ODC) was detected in the rat model. The whole experiments were to verify the promotion of LSSD to the development of tumor, and to reveal its mechanism.Methods The study were divided into three parts. And different animal model was set up in each part, including lump-bearing mouse model, ascitic fluid tumor-bearing mouse model and rat model of chemical hepatocarcinoma induced with diethylnitrosamine (DENA). In each part of the study, the laboratory animals were randomly divided into five groups, including the normal control, LSSD group, tumor group, integrated LSSD and tumor group, and the treatment group. Acting as a counterevidence, the treatment group was treated with Shuganjianpi Decoction which could eliminate the effect of LSSD factor. The changes of appearance, behavior, diet, stool and body weight of the animals were observed during the course of experiment. 10 days later, the spleens of mice in the lump-bearing model were picked off and T-lymphocyte proliferation and B-lymphocyte proliferation rate were detected by MTT colorimetry, IFN- y and IL-2 levels were detected by mitogen-activating lymphoblast assay and macrophage NCV production assay respectively, and the weight of the tumors and thymuses were measured. At the same time, the levels of IL-4 and IL-10 in serum were detected by ELISA. The change of Thl/Th2 cytokines reflects the influence of LSSD on Thl/Th2 immune balance of tumor-bearing mice. In the ascitic fluid tumor-bearing mouse model, the ascitic fluid cells of each group were extracted and the cell cycles of them was detected by flow cytometry. In the model of chemical hepatocarcinoma, the gene expression of ODC in rat livers were detected by reverse transcription-polymerase chain reaction (RT-PCR) in each group in the 9th, 14th and 19th respectively.Results The results indicated that LSSD could suppress the immune function in the mouse model of "integrated LSSD and tumor". T-lymphocyte proliferation rate and B-lymphocyte proliferation rate both decreased, thymus indexes were diminished. The secretion of IL-2 and IFN-y of spleen lymphocytes diminished, but the serum levels of EL-4 and IL-10 increased. Thus led to the shift of Thl/Th2 immune balance towards Th2. Meanwhile, LSSD also changed the cell cycles of ascitic fluid tumors in mice. It caused an increased S phase fraction of tumor cellsand the acceleration of cell division and proliferation. And thus to promote the growth of tumors. After the treatment of Shuganjianpi Decoction, the above indicators were all improved significantly. In the rat model of experimental heptocarcinoma induced with DENA, LSSD obviously enhanced the gene expression of ODC in rat livers. The effect became more and more predominant with the development of hepatocarcinoma and immobilization. It was ascending rapidly during the early stage. The expression of ODC decreased after the treatment of Shuganjianpi Decoction.Cone I us i on LSSD had a negative effect on the cellular immune function of tumor-bearing animals. It could decrease the Thl cytokines, increase the Th2 cytokines and promote the shift of Thl/Th2 balance towards Th2, thus to help the tumor cells to escape from the immune surveillance. LSSD could also change the cell cycle and enhance the gene expression of ODC in tumor cells, and thus to promote the development of tumor. |
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