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如何探討中藥有效成分對 基因調控的影響

如何探討中藥有效成分對 基因調控的影響. Gene Regulation. Stimulation. Inflammatory protein. ex. cytokine, LPS, stress. ex. TNF, IL-1, MCP-1, ICAM-1. Receptor. Membrane. Protein kinase phosphorylation. Signal transduction. (Western Blot). (ex. ERK, P38, JNK). Protein. (ELISA). Protein-DNA binding.

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如何探討中藥有效成分對 基因調控的影響

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  1. 如何探討中藥有效成分對基因調控的影響

  2. Gene Regulation Stimulation Inflammatory protein ex. cytokine, LPS, stress ex. TNF, IL-1, MCP-1, ICAM-1 Receptor Membrane Protein kinase phosphorylation Signal transduction (Western Blot) (ex. ERK, P38, JNK) Protein (ELISA) Protein-DNA binding Transcription factor *(EMSA) (ex. AP-1,NF-B,Egr-1) mRNA MCP-1 Nuclear (Northern Blot) AP-1 NF-B TF AP-1 NF-B Egr-1 CM98/03/07/94

  3. Electrophoretic Mobility Shift Assay (EMSA) • 研究DNA結合蛋白與其相關結合序列相互作用的方法 Protein / DNA (transcription factor) / (promoter region)

  4. EMSA • Nuclear protein • DNA probe • Native gel (non-denaturing agarose gel) • Autoradiography

  5. Promoter activity Promoter Reporter gene Luciferase Luciferase Lucoferin + ATP Light

  6. Regulation of atherosclerosis-related genes by herbal medicines : Wog inhibits MCP-1 and TF gene expressions

  7. Protocol for gene regulation • mRNA ― Northern Blot • mRNA stability ― Northern Blot • protein ― ELISA or Western Blot • promoter activity ― Luciferase activity • activation of transcription factor ― EMSA • signal transduction ― Western Blot

  8. Atherosclerosis a progressive disease characterized by the accumulation of lipids and fibrous elements in the large arteries • classic concept : as a disorder of lipid metabolism and deposition • current view : as a chronic inflammatory disease

  9. Three stages in the life history of an atheroma • Initiation : permeability/ adhesion/ migration • Progression : fatty-streak formation/ advanced lesion • Complication : plaques rupture

  10. Gene expression in the development of atherosclerosis Initiation Progression Complication MCP-1 MMP TF VCAM-1 ICAM-1

  11. 中醫的觀點： 《外科正宗》--- 厚味膏梁, 薰蒸臟腑, 丹石補藥 消灼腎水; 房勞過度, 氣竭津傷, 致使經絡阻塞, 氣血凝滯而發病。 屬中醫“脈痺” 和“脫疽”的範疇 血瘀證

  12. 粥狀動脈硬化症：慢性的發炎性疾病 • Initiation Progression Complication • MCP-1 MMP TF • VCAM-1 • ICAM-1 • 清熱藥：預防及治療粥狀動脈硬化症  • 辨證論治→非得有證才能治﹖

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