IL-17A suppresses the expression of bone resorption-related proteinases and osteoclast differentiation via IL-17RA or IL-17RC receptors in RAW264.7 cells
IL-17A suppresses the expression of bone resorption-related proteinases and osteoclast differentiation via IL-17RA or IL-17RC receptors in RAW264.7 cells
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DOI：
10.1016/j.biochi.2009.12.011
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发表时间：
2010-04-01
期刊：
影响因子：
3.9
通讯作者：
Maeno, Masao
中科院分区：
文献类型：
--
作者：
Kitami, Satoshi;Tanaka, Hideki;Maeno, Masao
白介素17(IL-17)是由活化的T细胞和中性粒细胞产生的，通过诱导“核因子-kappaB受体激活剂(RANKL)”的表达，通过成骨细胞刺激破骨细胞性骨吸收。然而，IL-17对破骨细胞前体向破骨细胞分化以及对破骨细胞功能的直接影响尚不清楚。因此，我们利用RAW264.7细胞研究了IL-17A对破骨细胞前体细胞分化的影响，以及对这些细胞中碳酸氢酶II(CA II)、组织蛋白酶K、基质金属蛋白酶-9(MMP9)、RANK、c-FMS和IL-17受体表达的影响。在加入或不加入0.1、1.0、10或50 ng/mLIL-17的条件下，加入可溶性RANKL培养10天。采用实时定量聚合酶链式反应(Real-time PCR)和免疫印迹法(Western Blotting)分别检测CA II、组织蛋白酶K和基质金属蛋白酶-9(MMP9)的mRNA和蛋白表达水平。实时定量聚合酶链式反应检测RANK、c-FMS和IL-17受体的mRNA表达水平。用抗酒石酸酸性磷酸酶(TRAP)染色检测破骨细胞分化程度。培养第5天出现TRAP阳性细胞，加入10和50 ng/mLIL-17A后第5天和第7天细胞数量减少。加入IL-17A后第5天和/或第7天组织蛋白酶K、基质金属蛋白酶9和c-FMS的表达明显减少，而CA II和1L-17受体(A型)的表达分别在第3天和第7天显著增加。加入IL-17A不影响RANK和IL-17受体(C型)的表达。这些结果表明，高浓度的IL-17A抑制了破骨细胞前体向破骨细胞的分化。此外，IL-17A通过减少破骨细胞中组织蛋白酶K和基质金属蛋白酶-9的产生，抑制骨吸收过程中基质蛋白的水解。(C)2009年爱思唯尔·马森公司。版权所有。
Interleukin-17 (IL-17) is produced exclusively by activated T cells and neutrophils, and stimulates osteoclastic bone resorption via osteoblasts by inducing the expression of "receptor activator of NF-kappa B (RANK) ligand" (RANKL). However, the direct effects of IL-17 on the differentiation of osteoclast precursors into osteoclasts and on the function of osteoclasts have not been clarified. Therefore, we examined the effects of IL-17A on the differentiation of osteoclast precursors using RAW264.7 cells and also on the expression of carbonic anhydrase II (CA II), cathepsin K, matrix metalloproteinases-9 (MMP-9), RANK, c-fms, and IL-17 receptors in these cells. The cells were cultured with or without 0.1, 1.0, 10 or 50 ng/mL IL-17 in the presence of soluble RANKL for up to 10 days. The CA II, cathepsin K, and MMP-9 mRNA and protein expression levels were examined using real-time PCR and Western blotting, respectively. The mRNA expression levels of RANK, c-fms, and IL-17 receptors were monitored by real-time PCR. Osteoclast differentiation was estimated using tartrate-resistant acid phosphatase (TRAP) staining of the cells. TRAP-positive cells were observed after day 5 of culture, and the number of cells decreased in the presence of 10 and 50 ng/mL IL-17A at days 5 and 7. In the presence of IL-17A, the expressions of cathepsin K, MMP-9 and c-fms decreased markedly on days 5 and/or 7 of culture, whereas the expression of CA II and 1L-17 receptor (type A) increased remarkably at days 3 and 7, respectively. The expression of RANK and IL-17 receptor (type C) was not affected by the addition of IL-17A. These results suggest that the differentiation of osteoclast precursors into osteoclasts is suppressed at high concentrations of IL-17A. Furthermore, IL-17A suppresses the hydrolysis of matrix proteins during bone resorption by decreasing the production of cathepsin K and MMP-9 in osteoclasts. (C) 2009 Elsevier Masson SAS. All rights reserved.