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

mRNAAP-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-1ex. 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 gene Luciferase

　　　　　　　　　 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﹖