Induction of Various Pro-inflammatory Mediators and Their Receptors in Human Astrocytoma Cell Line Stimulated by β-amyloid Protein

J Korean Neurol Assoc > Volume 20(5); 2002 > Article

Journal of the Korean Neurological Association 2002;20(5): 515-524.

"성상세포종에서 β-amyloid 단백 투여 후 염증매개체와 수용체의 유도"

한서영 , 최영숙 ·이광수 ·김상호

가톨릭대학교 의과대학 병리학교실,가톨릭대학교 의과대학 신경과학교실

Induction of Various Pro-inflammatory Mediators and Their Receptors in Human Astrocytoma Cell Line Stimulated by β-amyloid Protein

Seo young Han

"Department of Pathology, College of Medicine, The Catholic University of Korea, Department of Neurology, College of Medicine, The Catholic University of Korea"

Abstract

"Background : The induction of production and production inhibition of α1-antichymotrypsin (ACT), IL-1 α, IL-1 α receptor and macrophage inflammatory protein-1 (MIP-1) receptor in A β1-42 (A β)-stimulated U373MG cell, the human astrocytoma cell line, have never been reported. Methods : U373MG cells (1 ×10 6 cells in RPMI-1640 media) were incubated for overnight after administration of a single dose of 20 μM of A βor 0.5 ng/ml of TNF αor both. Actinomycin D (2.5 μM) or cycloheximide (2.5 μM) was also added to the cell suspension. Messenger RNA expression of ACT, IL-1 α, IL-1 αreceptor and MIP-1 receptor was measured by RT-PCR. Western blot was done and nitrocellulose paper was stained with anti-ACT and anti-GFAP antibody. NF κB activation after treatment of A βin U373MG cells was detected by electrophoretic mobility-shift assay. Results : A βand TNF αboth increased production of ACT in a dose-dependent manner. TNF αenhanced A β-induced mRNA had increases of ACT, IL-1 α, IL-1 αreceptor and MIP-1 α receptor. Activated NF κB was demonstrated in the A β, TNF α-stimulated U373MG cells. Actinomycin suppresses mRNA level of ACT and IL-1 αreceptor but cycloheximide inhibits the expression of ACT, IL-1 αand MIP-1 αreceptor. Conclusions : TNF αincreases synthesis of ACT, IL-1 α, IL-1 αreceptor and MIP-1 αreceptor in A β-stimulated astro-cyte, which, as a result, may contribute to the neuroinflammation of Alzheimer ’s disease.Key Words : Amyloid beta-protein, Alpha-1-antichymotrypsin, Interleukin-1, Macrophage inflammatory protein-1, Actinomycin, NF-kappa B"