J Korean Orthop Res Soc.
2010 Dec;13(2):80-87.
Glucocorticoid Mechanism of Inhibition of the Inflammatory Cells in Lumbar Intervertebral Disc Cells Stimulated by TNF-alpha Production of Nuclear Factor-kappaB
- Affiliations
-
- 1Department of Orthopedic surgery, Incheon St. Mary's hospital, College of Medicine, the Catholic University of Korea, Seoul, Korea.
- 2Department of Orthopedic surgery, Daejeon St. Mary's hospital, College of Medicine, the Catholic University of Korea, Seoul, Korea. kimtwins72@paran.com
Abstract
- PURPOSE
To analyze the action mechanism of NF-kappaB, IkappaB-alpha and effect of the Dexamethasone (DEXA) in mediating this inflammation, after stimulating cultured herniated intervertebral disc cells with TNF-alpha.
MATERIALS AND METHODS
After cultured human intervertebral disc cells passaged three times, they were divided into four groups: A control group (A), DEXA treatment group (B), TNF-alpha treated group (C), TNF-alpha and DEXA were treated at the same time (D). IL-6 and IL-1beta gene expression were measured with semi-quantitative RT-PCR. Western blot analysis was performed to measure protein expression of IkappaB-alpha in the above groups for 10 minutes, 1 hour, 2 hours. In addition, in order to explain the mechanism of NF-kappaB nuclear binding for each group, the nuclear amount of NF-kappaB binding in the nucleus is measured by EMSA.
RESULTS
In RT-PCR, expression of IL-6 and IL-1beta was greatest in group C, followed by group D, group A. IkappaB-alpha expression of the group treated with DEXA was not detected in Western blot results within 10 minutes. However, if stimulated by TNF-alpha, the DEXA was not inhibited of IkappaB-alpha concentration. After 1 hour and 2 hours, IkappaB-alpha levels were expressed by cells autonomously (autoregulatory induction). EMSA results expression levels in nuclear protein was maintained in accordance with protein expression.
CONCLUSIONS
Our study shows that DEXA inhibits the production of mediators such as inflammatory IL-6 and IL-1beta, however, may not inhibit the transcription of NF-kappaB stimulated by TNF-alpha.
- Full Text Links
-
- Actions
-
Cited
- CITED
-
- Close
- Share
-
- Similar articles
-
- Effects of Peroxisome Proliferator-Activated Receptor-gamma on the Production of Tumor Necrosis Factor-alpha in Stimulated Human Monocoytes
- Apigenin Inhibits Tumor Necrosis Factor-alpha-Induced Production and Gene Expression of Mucin through Regulating Nuclear Factor-Kappa B Signaling Pathway in Airway Epithelial Cells
- The Regulatory Mechanism of Src Familly Kinase in Lipopolysaccharide (LPS) induced HF-kappaB Activation Pathway
- Role of protein kinases on NF- kappaB activation and cell death in bovine cerebral endothelial cells
- The Role of NF-kappaB in the TNF-alpha-induced Apoptosis of Lung Cancer Cell Line
