Induction of Various Pro-inflammatory Mediators and Their Receptors in Human Astrocytoma Cell Line Stimulated by beta-amyloid Protein.
- Author:
Seo Young HAN
1
;
Young Sook CHOI
;
Kwang Soo LEE
;
Sang Ho KIM
Author Information
1. Department of Pathology, College of Medicine, The Catholic University of Korea, Korea. complt@cmc.cuk.ac.kr
- Publication Type:Original Article
- Keywords:
Amyloid beta-protein;
Alpha-1-antichymotrypsin;
Interleukin-1;
Macrophage inflammatory protein-1;
Actinomycin;
NF-kappa B
- MeSH:
Alzheimer Disease;
Amyloid beta-Peptides*;
Astrocytes;
Astrocytoma*;
Blotting, Western;
Cell Line*;
Collodion;
Cycloheximide;
Dactinomycin;
Humans*;
Interleukin-1;
Interleukin-1alpha;
Macrophage Inflammatory Proteins;
NF-kappa B;
RNA, Messenger
- From:Journal of the Korean Neurological Association
2002;20(5):515-524
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
BACKGROUND: The induction of production and production inhibition of alpha1-antichymotrypsin (ACT), IL-1 alpha, IL-1 alpha receptor and macrophage inflammatory protein-1 (MIP-1) receptor in A beta1-42 (A beta)-stimulated U373MG cell, the human astrocytoma cell line, have never been reported. METHODS: U373MG cells (1 x 10(6) cells in RPMI-1640 media) were incubated for overnight after administration of a single dose of 20 micro M of A beta or 0.5 ng/ml of TNF alpha or both. Actinomycin D (2.5 micro M) or cycloheximide (2.5 micro M) was also added to the cell suspension. Messenger RNA expression of ACT, IL-1 alpha, IL-1 alpha receptor 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 kappa B activation after treatment of A beta in U373MG cells was detected by electrophoretic mobility-shift assay. RESULTS: A beta and TNF alpha both increased production of ACT in a dose-dependent manner. TNF alpha enhanced A beta-induced mRNA had increases of ACT, IL-1 alpha, IL-1 alpha receptor and MIP-1 alpha receptor. Activated NF kappa B was demonstrated in the A beta, TNF alpha-stimulated U373MG cells. Actinomycin suppresses mRNA level of ACT and IL-1 alpha receptor but cycloheximide inhibits the expression of ACT, IL-1 alpha and MIP-1 alpha receptor. CONCLUSIONS: TNF alpha increases synthesis of ACT, IL-1 alpha, IL-1 alpha receptor and MIP-1 alpha receptor in A beta-stimulated astrocyte, which, as a result, may contribute to the neuroinflammation of Alzheimer's disease.
