| Author: | Wang, Yuen-yuen |
| Title: | Proteomic approach in the screening of immunomodulatory activities in traditional Chinese medicine |
| Degree: | M.Phil. |
| Year: | 2002 |
| Subject: | Hong Kong Polytechnic University -- Dissertations. Medicine, Chinese. Herbs -- Therapeutic use. Immunological adjuvants. |
| Department: | Department of Applied Biology and Chemical Technology |
| Pages: | xiii, 187 leaves : ill. ; 30 cm. |
| Language: | English |
| Abstract: | The proteomic approach has fast evolved as the method of choice for novel drug discovery. Today, many screening programs with this approach use transformed cell lines as the effectors for investigation. After exposing to the compound of interest, enhanced propagation of a cell line will denote prospective immunomodulatory activities. However, this "popular." approach does not account for the fact that the induction of a wide spectrum of immunomodulatory activities requires the presence and interactions of various types of immune cells including macrophages, B cells and T cells. More importantly, using this approach, multi-targeting effects will be difficult to detect. In our study, we used primary splenocytes as the effectors. Splenocytes (which consists of macrophages, B- and T-cells found in the spleen) are capable of inducing a full spectrum of immunomodulatory activities. On the other hand, two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) can be used to examine the concurrent expression of more than 1,000 different protein species. Therefore, multi-targeting effects of any drug under investigation through the up- or down-regulation of compounds can be detected conveniently. After confirming the immunomodulatory activities of ginsenoside Rgl on production of TNF-a and IFN-g using western blot and ELISA assay, comparison of 2D gel patterns between normal and Rgl treated samples was performed. Our results showed that 87 proteins were up-regulated, 38 were down-regulated, and 38 had no significant change. With the aid of MALDI-TOF-MS, 7 proteins were identified. For the identified proteins, the T cell surface glycoprotein CD5, DNA polymerase and homeotic protein HL-2 were up-regulated, and they are involved in the lymphocyte proliferation process. Cytochrome C oxidase and G protein are the other two identified up-regulated proteins, which are involved in the transmembrane signaling process and cytokine IL-2 induction process. Hypothetical anti-proliferative protein is the only identified protein that was down-regulated. It may be involved in the suppression of white cell proliferation. The role of another identified up-regulated protein, alpha-mannosidase II is not clear. The significance of this finding is currently unknown. Nevertheless, the exact immunomodulatory mechanism of Rgl can be easily demonstrated using this approach. Results of this study not only will be very useful in understanding immunomodulatory mechanisms of Rgl, this screening platform can also provide a framework and background for large-scale screening of TCM for immunomodulation as well as development of new drugs for cancer prevention and treatment. |
| Rights: | All rights reserved |
| Access: | open access |
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