Increase of TNFα-stimulated osteoarthritic chondrocytes apoptosis and decrease of matrix metalloproteinases 9 by NF-κB inhibition.
- Author:
Yan WANG
1
;
De Ling LI
;
Xin Bo ZHANG
;
Yuan Hui DUAN
;
Zhi Hong WU
;
Dong Sheng HAO
;
Bao Sheng CHEN
;
Gui Xing QIU
Author Information
- Publication Type:Journal Article
- Keywords: Apoptosis; Chondrocytes; Gelatinase; NF-κB; Tumor necrosis factor α
- MeSH: Aged; Apoptosis; drug effects; Caffeic Acids; pharmacology; therapeutic use; Calcium; physiology; Cells, Cultured; Chondrocytes; drug effects; enzymology; secretion; Drug Evaluation, Preclinical; Female; Humans; Matrix Metalloproteinase 2; metabolism; Matrix Metalloproteinase 9; metabolism; Middle Aged; NF-kappa B; antagonists & inhibitors; Osteoarthritis; drug therapy; enzymology; Phenylethyl Alcohol; analogs & derivatives; pharmacology; therapeutic use; Tumor Necrosis Factor-alpha; pharmacology
- From: Biomedical and Environmental Sciences 2013;26(4):277-283
- CountryChina
- Language:English
-
Abstract:
OBJECTIVETo investigate the in vitro effect of caffeic acid phenethyl ester (CAPE), a NF-κB inhibitor, on the apoptosis of osteoarthritic (OA) chondrocytes and on the regulation of the gelatinases matrix metalloproteinase 2 (MMP-2) and matrix metalloproteinase 9 (MMP-9).
METHODSAnnexin V-FITC/propidium iodide (PI) labeling and western blotting were used to observe and determine the apoptosis in TNFα-stimulated primary cultured osteoarthritic chondrocytes. Also, gelatin zymography was applied to examine MMP-2 and MMP-9 activities in supernatants.
RESULTSIt was confirmed by both flow cytometry and western blotting that chondrocytes from OA patients have an apoptotic background. Use of CAPE in combination with 10 ng/mL of TNFα for 24 h facilitated the apoptosis. MMP-9 in the supernatant could be autoactivated (from proMMP-9 to active MMP-9), and the physiologic calcium concentration (2.5 mmol/L) could delay the autoactivation of MMP-9. The activities of MMP-2 and MMP-9 in the fresh supernatant increased significantly in response to stimulation by 10 ng/mL of TNFα for 24 h. The stimulatory effect of TNFα just on proMMP-9 was counteracted significantly by CAPE.
CONCLUSIONNF-κB could prevent chondrocytes apoptosis though its activation was attributed to the increase of proMMP-9 activity induced by TNFα (a pro-apoptotic factor). Therefore, therapeutic NF-κB inhibitor was a 'double-edged swords' to the apoptosis of chondrocytes and the secretion of MMP-9.