碩士 / 國立宜蘭大學 / 生物技術研究所碩士班 / 97 / The shrimp anti-lipopolysaccharide factor peptide (anti-LPS), SALF, has been demonstrated to protect animals against sepsis. Gram-negative bacterial sepsis may be due to the activation of host immune responses by LPS. LPS may go through the MAPK or NFκB pathway to increase the level of the proinflammatory cytokines, such as tumor necrosis factor alpha (TNF-α), interleukin (IL)-6, IL-12, and IL-10, which may damage cells and lead to organ injury. The most important proinflammatory cytokine was TNF-α. RAW264.7 cell was treated with LPS (10 or 100 ng/ml) with different concentrations of SALF (0~100 μg/ml) for 4h, and the cell culture supernatants were screened for TNF-α by ELISA. The observation showed that SALF (20 μg/ml) was able to modulate the inflammatory responses by inhibiting the release of the TNF-α in LPS (100μg/ml)-stimulated mouse macrophage cells (RAW264.7). Treatment of SALF for 2h followed by LPS for 24h in RAW264.7 cells inhibited the TNF-α production around 79.1%. The microarray results showed that some genes had been modulated by two-fold changes. We selected the genes encoding immune-related proteins such as CD36, SCAR, ITGAX, CLU, ACTB, MYC, ANP32A, KLF2, CDC25L, SLPI, KS1, NOD1 and NFκB, which changed two-fold up or down for further analysis. Furthermore, we checked the expression of selected genes by real-time PCR in SALF and LPS treated RAW264.7 cells with the presence of specific inhibitors or activators to confirm the NFκB mediated mechanism. Finally, we found that SALF not only inhibited TNF-α, but also inhibited the cytokine of IL-1β and IL-6.
| Identifer | oai:union.ndltd.org:TW/097NIU07108007 |
| Date | January 2009 |
| Creators | Ming-Ching Lin, 林明靜 |
| Contributors | Shih-Bin Lin, Jyh-Yih Chen, 林世斌, 陳志毅 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 97 |
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