如何探討中藥有效成分對 基因調控的影響

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

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)

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

Promoter activity Promoter Reporter geneLuciferase Luciferase Lucoferin + ATP Light

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

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

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

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

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

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

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﹖