Study On Immunosuppressive And Anti-tumor Effects Of Different Extracts From Caesalpinia Sappan L

With the effects of various tumorigenic factors, cells of local tissues will lose control on their growth in the level of genes, which leads to clonal hyperplasia or insufficient apoptosis and results in a neoformation called tumor, often existing in the form of local mass. Whether the tumor is benign or malignant, or has its origins in epithelial tissue or mesenchymal tissue, it manifests abnormal proliferation of cells losing control. This abnormal growing ability can not only be seen from tumor's continuous growth but also from aggression to its adjacent normal tissues and transfer to other parts through blood vessels, lymphatic vessels and body cavity especially for malignant tumor, which is always the cause of death.In traditional Chinese medicine, physicians in past dynasties regarded that the pathogenesis of tumor mainly contained insufficient healthy qi, blood stasis and phlegm stagnation and invasion of pathogenic toxins etc. They thought tumor is closely related with blood stasis syndrome based on the clinical manifestations of tumor patients such as pathologic tumor mass, pain, hemorrhage, purple tongue with stasis or ecchymosis or blood spots, swarthy complexion and scaly skin in the advanced stage.In recent years, with constant study on molecular biology theory, the study of treating tumor with traditional Chinese medicine has made great progress, giving us a preliminary understanding of the molecule mechanism of inhibiting tumor growth, enhancing immunity and preventing the transfer of tumor with Chinese herbs. The study of traditional Chinese medicine's effects on tumor has developed from the preliminary study on immune function regulation to the current anti-tumor angiogenesis research and apoptosis induction and telomerase activity inhibition research.Hematoxylon (Caesalpinia Sappan L, CAE) is the dry heartwood of Leguminosae Caesalpinia which has the effect of promoting blood flow to eliminate stasis and dissolving the swelling to kill the pain. Caesalpinia Sappan L is used to treat dysmenorrheal, rheumatic pain and traumatic injuries and pains etc in Chinese medicine. It can also be used to treat tumor, as the modern pharmacological research shows that it has anti-tumor and immunosuppressive functions, although its specific anti-tumor mechanism is still unknown. Therefore, we can use modern medical testing technology to study the anti-tumor and immunosuppressive mechanism of Hematoxylin alcohol extracts.In the first part, the main purpose is to observe the effect of different extracts of Hematoxylon in mouse thymus, peripheral T cell proliferation and INF-γgeneration. Different extracts prepared from Caesalpinia Sappan L were lavaged to the mice in order to count thymus index by taking out their thymus, to calculate the proliferation capacity of spleen T cells by using MTT method, to measure the INF-γcontent generated by T cell culture supernatant by using double-antibody sandwich ELISA method. The results showed that different extracts of Hematoxylon not only had remarkable inhibition effect in thymus weight but also in T cell's proliferation and INF-γgeneration abilities. Hematoxylin alcohol and n-butanol extracts had stronger inhibiting effects on T cells.Given the strong effects of alcohol extract, in the second part of the study, the main purpose is to observe Hematoxylin alcoholl extract's cytotoxicity and apoptosis inducing effects on A549 cell of lung caner. Hematoxylin alcoholl extracts with different concentration were co-cultured with lung cancer A549 cells in order to observe effects on cell proliferation by using MTT method, study the effects on cytotoxicity and apoptosis of lung cancer A549 cells using optical microscope, phosphatidylserine ecstrophy analysis and cell period analysis. The results showed that Hematoxylin alcohol extracts had inhibiting effects on human lung cancer A549 cells in a dose-dependent manner; besides the cell apoptosis induced by these extracts, there existed other forms of cell death and affected cells in every period.Following the second part of anti-tumor experiment outside, here comes the internal one. The third part is mainly to observe the extract's anti-tumor effect on H22 ascites tumor cells. Tumor-bearing rat model was built by subcutaneous inoculation of H22 ascites tumor cells and lavaged with Hematoxylin alcohol extracts in order to observe the tumor growth conditionsmof tumor-bearing mice in each group, measure the tumor cell apoptotic index by using the nick end labeling method and apoptotic protein Bcl-2, Bax expression by using immunohistochemical technology. The results showed that Hematoxylin alcohol extract can inhibit the H22 transplantation tumor growth, facilitate tumor cell apoptosis, reduce Bcl-2/Bax ratio of tumor cells and the mechanism was related with the induction of tumor cell apoptosis by regulating the expression of apoptosis-related protein Bcl-2/Bax.