Lipopolysaccharide and proinflammatory cytokines stimulate interleukin-6 expression in C2C12 myoblasts:: role of the Jun NH2-terminal kinase

被引:72
作者
Frost, RA [1 ]
Nystrom, GJ [1 ]
Lang, CH [1 ]
机构
[1] Penn State Univ, Milton S Hershey Med Ctr, Coll Med, Dept Cellular & Mol Physiol, Hershey, PA 17033 USA
关键词
skeletal muscle; tumor necrosis factor-alpha; interleukin-1; beta; mitogen-activated protein kinase;
D O I
10.1152/ajpregu.00164.2003
中图分类号
Q4 [生理学];
学科分类号
071003 ;
摘要
IL-6 is a major inflammatory cytokine that plays a central role in coordinating the acute-phase response to trauma, injury, and infection in vivo. Although IL-6 is synthesized predominantly by macrophages and lymphocytes, skeletal muscle is a newly recognized source of this cytokine. IL-6 from muscle spills into the circulation, and blood-borne IL-6 can be elevated > 100-fold due to exercise and injury. The purpose of the present study was to determine whether inflammatory stimuli, such as LPS, TNF-alpha, and IL-1beta, could increase IL-6 expression in skeletal muscle and C2C12 myoblasts. Second, we investigated the role of mitogen-activated protein ( MAP) kinases, and the Jun NH2-terminal kinase (JNK) in particular, as a mediator of this response. Intraperitoneal injection of LPS in mice increased the circulating concentration of IL-6 from undetectable levels to 4 ng/ml. LPS also increased IL-6 mRNA 100-fold in mouse fast-twitch skeletal muscle. Addition of LPS, IL-1beta, or TNF-alpha directly to C2C12 myoblasts increased IL-6 protein (6- to 8-fold) and IL-6 mRNA (5- to 10-fold). The response to all three stimuli was completely blocked by the JNK inhibitor SP-600125 but not as effectively by other MAP kinase inhibitors. SP-600125 blocked LPS-stimulated IL-6 synthesis dose dependently at both the RNA and protein level. SP-600125 was as effective as the synthetic glucocorticoid dexamethasone at inhibiting IL-6 expression. SP-600125 inhibited IL-6 synthesis when added to cells up to 60 min after LPS stimulation, but its inhibitory effect waned with time. LPS stimulated IL-6 mRNA in both myoblasts and myotubes, but myoblasts showed a proportionally greater LPS-induced increase in IL-6 protein expression compared with myotubes. SP-600125 and the proteasomal inhibitor MG-132 blocked LPS-induced degradation of IkappaB-alpha/epsilon and LPS-stimulated expression of IkappaB-alpha mRNA. Yet, only SP-600125 and not MG-132 blocked LPS-induced IL-6 mRNA expression. This suggests that IL-6 gene expression is a downstream target of JNK in C2C12 myoblasts.
引用
收藏
页码:R1153 / R1164
页数:12
相关论文
共 56 条
[1]   HUMAN INTERLEUKIN-6 AND TUMOR NECROSIS FACTOR-ALPHA PRODUCTION STUDIED AT A SINGLE-CELL LEVEL [J].
ANDERSSON, U ;
MATSUDA, T .
EUROPEAN JOURNAL OF IMMUNOLOGY, 1989, 19 (06) :1157-1160
[2]   Chronic inflammation and the effect of IGF-I on muscle strength and power in older persons [J].
Barbieri, M ;
Ferrucci, L ;
Ragno, E ;
Corsi, A ;
Bandinelli, S ;
Bonafè, M ;
Olivieri, F ;
Giovagnetti, S ;
Franceschi, C ;
Guralnik, JM ;
Paolisso, G .
AMERICAN JOURNAL OF PHYSIOLOGY-ENDOCRINOLOGY AND METABOLISM, 2003, 284 (03) :E481-E487
[3]   In vivo expression of proinflammatory mediators in the adult heart after endotoxin administration: the role of toll-like receptor-4 [J].
Baumgarten, G ;
Knuefermann, P ;
Nozaki, N ;
Sivasubramanian, N ;
Mann, DL ;
Vallejo, JG .
JOURNAL OF INFECTIOUS DISEASES, 2001, 183 (11) :1617-1624
[4]   SP600125, an anthrapyrazolone inhibitor of Jun N-terminal kinase [J].
Bennett, BL ;
Sasaki, DT ;
Murray, BW ;
O'Leary, EC ;
Sakata, ST ;
Xu, WM ;
Leisten, JC ;
Motiwala, A ;
Pierce, S ;
Satoh, Y ;
Bhagwat, SS ;
Manning, AM ;
Anderson, DW .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2001, 98 (24) :13681-13686
[5]   Purification of mouse primary myoblasts based on α7 integrin expression [J].
Blanco-Bose, WE ;
Yao, CC ;
Kramer, RH ;
Blau, HM .
EXPERIMENTAL CELL RESEARCH, 2001, 265 (02) :212-220
[6]   Eccentric exercise markedly increases c-Jun NH2-terminal kinase activity in human skeletal muscle [J].
Boppart, MD ;
Aronson, D ;
Gibson, L ;
Roubenoff, R ;
Abad, LW ;
Bean, J ;
Goodyear, LJ ;
Fielding, RA .
JOURNAL OF APPLIED PHYSIOLOGY, 1999, 87 (05) :1668-1673
[7]   Restraint of proinflammatory cytokine biosynthesis by mitogen-activated protein kinase phosphatase-1 in lipopolysaccharide-stimulated macrophages [J].
Chen, PL ;
Li, J ;
Barnes, J ;
Kokkonen, GC ;
Lee, JC ;
Liu, YS .
JOURNAL OF IMMUNOLOGY, 2002, 169 (11) :6408-6416
[8]   Quinone reductase inhibitors block SAPK/JNK and NFκB pathways and potentiate apoptosis [J].
Cross, JV ;
Deak, JC ;
Rich, EA ;
Qian, YY ;
Lewis, M ;
Parrott, LA ;
Mochida, K ;
Gustafson, D ;
Vande Pol, S ;
Templeton, DJ .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1999, 274 (44) :31150-31154
[9]   Interleukin 6 causes growth impairment in transgenic mice through a decrease in insulin-like growth factor-I - A model for stunted growth in children with chronic inflammation [J].
DeBenedetti, F ;
Alonzi, T ;
Moretta, A ;
Lazzaro, D ;
Costa, P ;
Poli, V ;
Martini, A ;
Ciliberto, G ;
Fattori, E .
JOURNAL OF CLINICAL INVESTIGATION, 1997, 99 (04) :643-650
[10]   INSULIN-STIMULATED MUSCLE GLUCOSE CLEARANCE IN PATIENTS WITH NIDDM - EFFECTS OF ONE-LEGGED PHYSICAL-TRAINING [J].
DELA, F ;
LARSEN, JJ ;
MIKINES, KJ ;
PLOUG, T ;
PETERSEN, LN ;
GALBO, H .
DIABETES, 1995, 44 (09) :1010-1020