Inducible hydrogen sulfide synthesis in chondrocytes and mesenchymal progenitor cells: is H2S a novel cytoprotective mediator in the inflamed joint?
Inducible hydrogen sulfide synthesis in chondrocytes and mesenchymal progenitor cells: is H2S a novel cytoprotective mediator in the inflamed joint?
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DOI：
10.1111/j.1582-4934.2011.01357.x
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发表时间：
2012-04
影响因子：
5.3
通讯作者：
Whiteman M
中科院分区：
文献类型：
--
作者：
Fox B;Schantz JT;Haigh R;Wood ME;Moore PK;Viner N;Spencer JP;Winyard PG;Whiteman M
硫化氢(H_2S)最近被认为是一种内源性炎症介质，存在于人类滑液中。本研究检测了原代培养的人关节软骨细胞(HAC)和间充质祖细胞(MPC)能否在与人类关节疾病相关的促炎细胞因子的作用下合成H_2S，以及细胞对内源性和药物性H_2S的反应。用IL-1β、IL-6、肿瘤坏死因子-α和脂多糖刺激HAC和MPC。用Western印迹和锌离子捕集分光光度法分别检测硫化氢合成酶CBS和Cse的表达和酶活性。用过氧化氢、过氧亚硝酸盐供体SIN-1和4-羟基壬烯醛(4-HNE)诱导细胞氧化应激。用3-(4，5-二甲基-2-基)-2，5-二苯基四氮唑(MTT法)和乳酸脱氢酶(LDH)法检测细胞死亡。用流式细胞仪原位检测线粒体膜电位(DCM)。内源性H_2S的合成分别被小干扰RNA介导的CSE和CBS以及药物抑制剂D、L-丙氨酸甘氨酸和氨基乙酸酯所抑制。利用GYY4137产生外源硫化氢。在基础条件下，HAC和MPC表达CBS和CSE，并以CBS依赖的方式合成H_2S，而IL-1β、肿瘤坏死因子-α、IL-6或内毒素诱导细胞表达CsE和活性。GYY4137处理可显著抑制氧化应激诱导的细胞死亡，但药物抑制硫化氢合成或CBS/CSE-siRNA处理可增加氧化应激诱导的细胞死亡。这些数据表明，CSE是培养的HAC和MPC中硫化氢的诱导源。硫化氢可能代表了炎症关节中一种新的内源性细胞保护机制，提示了治疗干预的潜在机会。
Hydrogen sulfide (H2S) has recently been proposed as an endogenous mediator of inflammation and is present in human synovial fluid. This study determined whether primary human articular chondrocytes (HACs) and mesenchymal progenitor cells (MPCs) could synthesize H2S in response to pro-inflammatory cytokines relevant to human arthropathies, and to determine the cellular responses to endogenous and pharmacological H2S. HACs and MPCs were exposed to IL-1β, IL-6, TNF-α and lipopolysaccharide (LPS). The expression and enzymatic activity of the H2S synthesizing enzymes cystathionine-β-synthase (CBS) and cystathionine-γ-lyase (CSE) were determined by Western blot and zinc-trap spectrophotometry, respectively. Cellular oxidative stress was induced by H2O2, the peroxynitrite donor SIN-1 and 4-hydroxynonenal (4-HNE). Cell death was assessed by 3-(4,5-dimethyl-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and lactate dehydrogenase (LDH) assays. Mitochondrial membrane potential (DCm) was determined in situ by flow cytometry. Endogenous H2S synthesis was inhibited by siRNA-mediated knockdown of CSE and CBS and pharmacological inhibitors D,L-propargylglycine and aminoxyacetate, respectively. Exogenous H2S was generated using GYY4137. Under basal conditions HACs and MPCs expressed CBS and CSE and synthesized H2S in a CBS-dependent manner, whereas CSE expression and activity was induced by treatment of cells with IL-1β, TNF-α, IL-6 or LPS. Oxidative stress-induced cell death was significantly inhibited by GYY4137 treatment but increased by pharmacological inhibition of H2S synthesis or by CBS/CSE-siRNA treatment. These data suggest CSE is an inducible source of H2S in cultured HACs and MPCs. H2S may represent a novel endogenous mechanism of cytoprotection in the inflamed joint, suggesting a potential opportunity for therapeutic intervention.
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影响因子：
4.8
作者：
Chiku, Taurai;Padovani, Dominique;Banerjee, Ruma
通讯作者：
Banerjee, Ruma
影响因子：
20.1
作者：
Calvert JW;Jha S;Gundewar S;Elrod JW;Ramachandran A;Pattillo CB;Kevil CG;Lefer DJ
通讯作者：
Lefer DJ
影响因子：
37.8
作者：
Calvert JW;Elston M;Nicholson CK;Gundewar S;Jha S;Elrod JW;Ramachandran A;Lefer DJ
通讯作者：
Lefer DJ
影响因子：
6.7
作者：
Asada, S;Fukuda, K;Hamanisi, C
通讯作者：
Hamanisi, C
影响因子：
4.4
作者：
Blanco, FJ;López-Armada, MJ;Maneiro, E
通讯作者：
Maneiro, E