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題 名 | 以系統生物學研究平台探討中草藥防治敗血症之病理生理與治療機轉--以靈芝為例,結合基因體學、蛋白質體學與生物資訊學之研究=Evaluation of Pathophysiological and Therapeutic Mechanisms of Herbal Medicines in the Prevention and Treatment of Sepsis by Systems Biology Research Platform: Studies on Ganoderma lucidum with a Combination of Genomics, Proteomics and Bioin formatics |
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作 者 | 汪貴珍; | 書刊名 | 中醫藥年報 |
卷 期 | 29:8 2011.09[民100.09] |
頁 次 | 頁93-129 |
專 輯 | 中醫藥基因體相關研究 |
分類號 | 414.33 |
關鍵詞 | 系統生物學; 基因體學; 蛋白質體學; 生物資訊學; 敗血症; 新藥開發; Systems biology; Genomics; Proteomics; Bioinformaticsm; Sepsis; Therapeutic drug development; |
語 文 | 中文(Chinese) |
中文摘要 | 敗血症是因感染而引起的複雜發炎反應,具高罹患率與死亡率;研發安全性高的防治藥物,實刻不容緩。本計畫利用系統生物學¾基因體學、蛋白質體學及生物資訊學,以細胞與動物模型配合生化與病理藥理學分析,建立並驗證敗血症之系統生物平台與網絡建構,同時研究中草藥防治敗血症之分子、細胞到活體動物各層面的系統性藥理機制。運用大鼠cDNA 微陣列、二維電泳膠、介質輔助雷射脫附/游離飛行時間式質譜儀,以及細胞激素蛋白質陣列等分析,透過生物資訊學資料庫:生物訊息路徑(BioCarta)、生物代謝路徑(KEGG)及基因註解與分類(Gene Ontology)進行注解;並以西方墨點法、qRT-PCR、siRNA 及microRNA 等實驗,建立並驗證特定的訊息傳遞或功能路徑。 實驗結果顯示血管內皮細胞受到細菌內毒素刺激後,直接或間接改變NF-κB上游Pias1 基因及SPA6 蛋白質的活性,及其下游基因(IL-1RN、IL-1β 及CD4)及蛋白質(IL-6 及MCP-1)的表現;並引起抗發炎(IL-1RN)、抗細胞程式性死亡(Rbpsun)及抗氧化(Mpo、PRDX1、SODM、FRIH 及FRIL1)基因及蛋白質的表現。針對特定標的基因與蛋白質研究,發現靈芝、肝炎草、厚葉鐵線蓮、金絲草等中草藥成分具有保護作用。其中,靈芝的部分純化物具有雙向的免疫調節作用,肝炎草與金絲草之抗發炎活性成分為新化合物,厚葉鐵線蓮成分具選擇性抗炎作用。 |
英文摘要 | Sepsis describes a complex inflammatory response with high incidence and mortality that results from the infection and innate immune activation. Systems biology, including microarray, proteomics and bioinformatics, was used to establish the pathophysiological platform. In the present study, the methods, including cDNA microarray, 2-DE and MALDI-TOF MS/MS, cytokine protein array and bioinformatic tools, combining with BioCarta, KEGG and Gene Ontology, were used to establish the systemic platform and the specific molecular detecting markers of sepsis. The effects of herbal medicines on gene and protein network were also examined by Western blotting, qRT-PCR, siRNA, microRNA, ELISA and other molecular biology analysis in in vivo and in vitro models. In conclusion, the results showed that the proinflammatory cytokines (IL-1b and IL-6) and chemokine (MCP-1) can be up-regulated by lipopolysaccharide (LPS) through the activation of the NF-kB signaling and transcription regulators (SPA6 and Pias1) which can also regulate the activity of the NF-kB signaling pathway. Interestingly, the endothelial cells can also up-regulate the mediators of anti-inflammation (IL-1RN), antiapoptosis (Rbpsun), and antioxidation (Mpo, PRDX1, SODM, FRIH, and FRIL1) to protect themselves against LPS treatment. The bioactive components have been isolated and identified from Ganoderma lucidum, Murdannia bracteata, Clematis crassifolia, Pogonatherum crinitum etc. Among them, G. lucidum produced two-way immunomodulation in macrophages. The naturally new ingredients from M. bracteata, C. crassifoliaand P. crinitum have potential in the prevention and treatment of disorders caused by inflammatory diseases. |
本系統中英文摘要資訊取自各篇刊載內容。