Induction of nerve growth factor expression and release by mechanical and inflammatory stimuli in chondrocytes: possible involvement in osteoarthritis pain.
Induction of nerve growth factor expression and release by mechanical and inflammatory stimuli in chondrocytes: possible involvement in osteoarthritis pain.
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DOI：
10.1186/ar4443
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发表时间：
2014-01-20
影响因子：
4.9
通讯作者：
Houard X
中科院分区：
文献类型：
--
作者：
Pecchi E;Priam S;Gosset M;Pigenet A;Sudre L;Laiguillon MC;Berenbaum F;Houard X
神经生长因子(NGF)水平在骨关节炎(OA)关节中升高，并与骨关节炎相关的疼痛有关。在软骨细胞中刺激NGF的刺激物是未知的。我们研究了机械应力和促炎细胞因子是否会影响软骨细胞NGF的合成。分别用增加IL-1β、前列腺素E_2、内脂素/烟酰胺磷酸核糖基转移酶或循环机械加压(0.5 MHz，1 Mpa)刺激原代培养的人骨关节炎软骨细胞、新生小鼠关节软骨细胞或软骨移植块。在刺激前，软骨细胞用NAMPT酶活性的特异性抑制剂消炎痛Apo866预处理，或通过靶向visfatin/NAMPT的siRNA转染组。用实时荧光定量聚合酶链式反应检测培养上清液中NGF的表达水平，用ELISA法测定培养上清液中NGF的释放。未受刺激的人和小鼠关节软骨细胞表达低水平的神经生长因子(人和小鼠关节软骨细胞和肋骨组织分别为19.2 ± 8.7pg/m l，13.5 ± 1.0m g/m l和4.4 ± 0.8m g/m l/mg组织)。机械应力可诱导条件培养液中NGF的释放。在IL-1β或内脂素/NAMPT(一种由软骨细胞产生的促炎症脂肪因子)刺激下，人和小鼠软骨细胞条件培养液中NGF mRNA的表达和NGF的释放均呈剂量依赖性增加。Visfatin/NAMPT也是一种胞内酶，作为NAD产生的限速酶。Apo866可抑制内脂素/NAMPT诱导的神经生长因子的表达，而靶向内脂素/NAMPT的小干扰RNA不能改变IL-1β介导的神经生长因子的表达。有趣的是，软骨细胞在IL-1β和Visfatin/NAMPT的作用下产生的前列腺素E_2并没有刺激神经生长因子的产生。环氧合酶抑制剂吲哚美辛始终不能抑制IL-1β诱导的神经生长因子的产生。这些结果表明，机械应力、IL-1β和细胞外内脂素/NAMPT均能刺激软骨细胞表达和释放神经生长因子，提示内脂素/NAMPT和IL-1β在骨性关节炎关节中的过度表达和软骨机械负荷的增加可能通过刺激软骨细胞表达和释放神经生长因子来介导骨性关节炎疼痛。
Nerve growth factor (NGF) level is increased in osteoarthritis (OA) joints and is involved in pain associated with OA. Stimuli responsible for NGF stimulation in chondrocytes are unknown. We investigated whether mechanical stress and proinflammatory cytokines may influence NGF synthesis by chondrocytes. Primary cultures of human OA chondrocytes, newborn mouse articular chondrocytes or cartilage explants were stimulated by increasing amounts of IL-1β, prostaglandin E2 (PGE2), visfatin/nicotinamide phosphoribosyltransferase (NAMPT) or by cyclic mechanical compression (0.5 Hz, 1 MPa). Before stimulation, chondrocytes were pretreated with indomethacin, Apo866, a specific inhibitor of NAMPT enzymatic activity, or transfected by siRNA targeting visfatin/NAMPT. mRNA NGF levels were assessed by real-time quantitative PCR and NGF released into media was determined by ELISA. Unstimulated human and mouse articular chondrocytes expressed low levels of NGF (19.2 ± 8.7 pg/mL, 13.5 ± 1.0 pg/mL and 4.4 ± 0.8 pg/mL/mg tissue for human and mouse articular chondrocytes and costal explants, respectively). Mechanical stress induced NGF release in conditioned media. When stimulated by IL-1β or visfatin/NAMPT, a proinflammatory adipokine produced by chondocytes in response to IL-1β, a dose-dependent increase in NGF mRNA expression and NGF release in both human and mouse chondrocyte conditioned media was observed. Visfatin/NAMPT is also an intracellular enzyme acting as the rate-limiting enzyme of the generation of NAD. The expression of NGF induced by visfatin/NAMPT was inhibited by Apo866, whereas IL-1β-mediated NGF expression was not modified by siRNA targeting visfatin/NAMPT. Interestingly, PGE2, which is produced by chondrocytes in response to IL-1β and visfatin/NAMPT, did not stimulate NGF production. Consistently, indomethacin, a cyclooxygenase inhibitor, did not counteract IL-1β-induced NGF production. These results show that mechanical stress, IL-1β and extracellular visfatin/NAMPT, all stimulated the expression and release of NGF by chondrocytes and thus suggest that the overexpression of visfatin/NAMPT and IL-1β in the OA joint and the increased mechanical loading of cartilage may mediate OA pain via the stimulation of NGF expression and release by chondrocytes.
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影响因子：
1.9
作者：
Alstergren, P;Ernberg, M;Kopp, S
通讯作者：
Kopp, S
影响因子：
3.7
作者：
Berenbaum, F;Jacques, C;Masliah, J
通讯作者：
Masliah, J
影响因子：
--
作者：
Evans, Laura;Williams, Anwen S.;Nowell, Mari
通讯作者：
Nowell, Mari
影响因子：
4.2
作者：
FRIEDMAN, WJ;ALTIOK, N;PERSSON, H
通讯作者：
PERSSON, H
DOI：
10.1152/ajpendo.00008.2005
发表时间：
2005-07-01
影响因子：
5.1
作者：
Bulló, M;Peeraully, MR;Trayhurn, P
通讯作者：
Trayhurn, P