title: Regulation of p38 MAPK phosphorylation inhibits chondrocyte apoptosis in response to heat stress or mechanical stress.
abstract: Activation of p38 MAPK has been associated with a stress response and with apoptotic processes. However, the function of p38 MAPK in chondrocytes is not clearly understood. In this study, we analyzed the expression of p38 MAPK in chondrocytes and investigated the function of p38 MAPK in response to heat stress and mechanical stress. Chondrocytes were isolated from human cartilage and cultured. Expression of p38 and phosphorylated p38 in cartilage of patients with osteoarthritis (OA) was compared to those in normal cartilage by immunohistochemistry and Western blotting. Human knee chondrocytes were exposed to heat stress or mechanical stress. Normal knee chondrocytes were pre-treated with SB203580 or p38 small interfering RNA (siRNA) before induction of heat stress or mechanical stress. Chondrocyte apoptosis was detected by TUNEL staining and Western blotting of cleaved caspases. OA and normal chondrocytes expressed p38; however, OA chondrocytes showed much higher phosphorylated p38 compared to normal chondrocytes. Heat stress or mechanical stress induced apoptosis and increased phosphorylated p38 in normal chondrocytes. The TUNEL positive cells and expression levels of phosphorylated p38 in response to stress decreased when chondrocytes were incubated with SB203580 or transfected with siRNA against p38. In conclusion, we have demonstrated that heat stress or mechanical stress increased chondrocyte apoptosis via phosphorylation of p38. Stress-induced chondrocyte apoptosis decreased due to inhibition of p38 MAPK activation. In contrast, the phosphorylation of p38 MAPK increased in OA chondrocytes. Our results show that down-regulation of p38 MAPK activation inhibits chondrocyte death induced by heat stress or mechanical stress.
authors.affiliation: Department of Orthopaedic Surgery, Kobe University Graduate School of Medicine, Kobe, Hyogo 670-0017, Japan.
authors.forename: Ken
authors.initials: K
authors.lastname: Takebe
authors.forename: Takayuki
authors.initials: T
authors.lastname: Nishiyama
authors.forename: Shinya
authors.initials: S
authors.lastname: Hayashi
authors.forename: Shingo
authors.initials: S
authors.lastname: Hashimoto
authors.forename: Takaaki
authors.initials: T
authors.lastname: Fujishiro
authors.forename: Noriyuki
authors.initials: N
authors.lastname: Kanzaki
authors.forename: Kohei
authors.initials: K
authors.lastname: Kawakita
authors.forename: Kenjiro
authors.initials: K
authors.lastname: Iwasa
authors.forename: Ryosuke
authors.initials: R
authors.lastname: Kuroda
authors.forename: Masahiro
authors.initials: M
authors.lastname: Kurosaka
mesh_terms: D017209:Apoptosis; D002478:Cells, Cultured; D019902:Chondrocytes / Q000201:enzymology* / Q000473:pathology; D015536:Down-Regulation / Q000187:drug effects / Q000235:genetics; D004789:Enzyme Activation / Q000187:drug effects / Q000235:genetics; D004791:Enzyme Inhibitors / Q000494:pharmacology; D015971:Gene Expression Regulation, Enzymologic; D018869:Heat-Shock Response; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology; D020370:Osteoarthritis, Knee / Q000201:enzymology* / Q000235:genetics / Q000473:pathology; D010766:Phosphorylation / Q000187:drug effects; D011725:Pyridines / Q000494:pharmacology; D034741:RNA, Small Interfering / Q000235:genetics; D048051:p38 Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*
chemical_list: D004791:Enzyme Inhibitors; D007093:Imidazoles; D011725:Pyridines; D034741:RNA, Small Interfering; D048051:p38 Mitogen-Activated Protein Kinases; C093642:SB 203580