Effect of hydrogen sulfide sources on inflammation and catabolic markers on interleukin 1β-stimulated human articular chondrocytes
Effect of hydrogen sulfide sources on inflammation and catabolic markers on interleukin 1β-stimulated human articular chondrocytes
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DOI：
10.1016/j.joca.2014.04.031
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发表时间：
2014-07-01
影响因子：
7
通讯作者：
Blanco, F. J.
中科院分区：
文献类型：
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作者：
Burguera, E. F.;Vela-Anero, A.;Blanco, F. J.
目的：硫化氢(H_2S)是继NO、CO之后的第三种气体递质，是一种炎症调节物质。为了验证其在骨性关节炎(OA)治疗中的应用价值，我们观察了两种外源性过氧化氢NASH和GYY4137对骨性关节炎炎症和分解代谢标志物的影响。用促炎症细胞因子(IL-1β，IL-1β，5 ng/ml)刺激细胞，并检测两种硫化氢来源改善其对细胞影响的能力。通过Griess反应定量一氧化氮(NO)的产生。免疫细胞化学方法检测诱导型一氧化氮合酶(NOS2)和基质金属蛋白酶13(MMP13)的蛋白表达。用定量逆转录-聚合酶链式反应(qRT-PCR)定量检测相关基因的表达。用特异性EIA法检测前列腺素E-2(PGE-2)、白介素6(IL-6)和基质金属蛋白酶13(MMP13)水平。结果：两种硫化氢来源细胞释放的NO、PGE-2、IL-6和MMP13在蛋白水平均显著降低。这是通过下调参与这些分子合成途径的相关基因，即NOS2、环氧合酶-2(COX2)、前列腺素E合成酶(PTGES)、IL-6和MMP13实现的。结论：NASH和GYY4137与IL1β激活的骨关节炎CHS联合应用，具有抗炎和抗分解代谢的作用。补充外源性硫化氢可以调节骨性关节炎发病和发展过程中相关基因的表达，部分通过减少核因子-kappaB的激活来对抗导致软骨破坏的IL-1β促炎信号。(C)2014国际骨性关节炎研究会。爱思唯尔有限公司出版。保留所有权利。
Objective: Hydrogen sulfide (H2S), the third gasotransmitter together with NO and CO, is emerging as a regulator of inflammation. To test if it might offer therapeutic value in the treatment of osteoarthritis (OA) we evaluated the effects of two exogenous sources of H2S, NaSH and GYY4137, on inflammation and catabolic markers that characterize OA.Method: Human chondrocytes (CHs) were isolated from OA tissue. Cells were stimulated with a pro-inflammatory cytokine (interleukin-1 beta, IL1 beta, 5 ng/ml) and the ability of the two H2S sources to ameliorate its effects on the cells was tested. Nitric oxide (NO) production was quantified through the Griess reaction. Protein levels of inducible NO synthase (NOS2) and matrix metalloproteinase 13 (MMP13) were visualized through immunocytochemistry (ICC). Relative mRNA expression was quantified with qRT-PCR. Prostaglandin-2 (PGE-2), interleukin 6 (IL6) and MMP13 levels were measured with specific EIAs. NF kappa B nuclear translocation was visualized with immunofluorescence.Results: Both H2S sources led to significant reductions in NO, PGE-2, IL6 and MMP13 released by the cells and at the protein level. This was achieved by downregulation of relevant genes involved in the synthesis routes of these molecules, namely NOS2, cyclooxigenase-2 (COX2), prostaglandin E synthase (PTGES), IL6 and MMP13. NF kappa B nuclear translocation was also reduced.Conclusion: NaSH and GYY4137 show anti-inflammatory and anti-catabolic properties when added to IL1 beta, activated osteoarthritic CHs. Supplementation with exogenous H2S sources can regulate the expression of relevant genes in OA pathogenesis and progression, counteracting IL1 beta pro-inflammatory signals that lead to cartilage destruction in part by reducing NF kappa B activation. (C) 2014 Osteoarthritis Research Society International. Published by Elsevier Ltd. All rights reserved.