Retinoid X receptor and peroxisome proliferator-activated receptor-gamma agonists cooperate to inhibit matrix metalloproteinase gene expression
Retinoid X receptor and peroxisome proliferator-activated receptor-gamma agonists cooperate to inhibit matrix metalloproteinase gene expression
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DOI：
10.1186/ar2564
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发表时间：
2008-01-01
影响因子：
4.9
通讯作者：
Brinckerhoff, Constance E.
中科院分区：
文献类型：
--
作者：
Burrage, Peter S.;Schmucker, Adam C.;Brinckerhoff, Constance E.
前言：我们最近研究了维甲酸X受体(RXR)配体LG100268(LG268)抑制白介素1-β(IL-1-β)诱导的软骨肉瘤细胞中基质金属蛋白酶-1(MMP1)和MMP13基因表达的能力。其他研究人员已经在罗格列酮处理的软骨细胞中证明了类似的效果，罗格列酮是过氧化物酶体增殖物激活受体-伽马(PPAR-Gamma)的配体，RXR是其专有的二聚化伙伴。本研究的目的是评价RXR和PPARγ配体联合作用对IL-1β诱导的软骨肉瘤细胞MMP1和MMP13表达的抑制作用，并探讨其分子机制。方法采用实时定量逆转录聚合酶链式反应检测LG268和罗格列酮对IL-1β诱导的Sw-1353软骨肉瘤细胞MMP1和MMP13基因转录的抑制作用。体外胶原蛋白破坏试验是对基质金属蛋白酶胶原酶活性的功能性读数。荧光素酶报告实验检测了一个可能的调控元件在基质金属蛋白酶-1和基质金属蛋白酶-13的启动子中的功能，染色质免疫沉淀(ChIP)检测了PPAR-γ和组蛋白乙酰化的变化。结果罗格列酮在体外可抑制IL-1-β诱导的Sw-1353细胞中基质金属蛋白酶-1和基质金属蛋白酶-13的表达，并抑制IL-1-β诱导的胶原破坏。联合应用LG268和罗格列酮对基质金属蛋白酶-1和基质金属蛋白酶-13的转录和胶原降解有相加的抑制作用。IL-1-β可抑制荧光素酶的表达，但罗格列酮和LG268对此无影响。CHIP显示，用IL-1-β处理，而不是LG268和罗格列酮处理，增加了两个MMP近端启动子的PPARγ。最后，罗格列酮或LG268可诱导靶受体及其结合配对的交叉SUMO化，IL-1-β单独作用对RXR和PPARγ的SUMO化无影响，但可拮抗配体诱导的两者的SUMO化。结论PPARγ和RXR的配体罗格列酮和LG268可能通过相似的机制抑制MMP1和MMP13的转录。联合治疗激活RXR：PPAR伽马异源二聚体的每个配对，并抑制IL-1-β诱导的基质金属蛋白酶-1和基质金属蛋白酶-13的表达，比单独使用任何一种化合物都更有效。我们的结论是，每种药物较低剂量的联合治疗效果可能会将这些化合物治疗的潜在副作用降至最低。
Introduction We recently described the ability of retinoid X receptor (RXR) ligand LG100268 (LG268) to inhibit interleukin-1- beta (IL-1-beta)-driven matrix metalloproteinase-1 (MMP-1) and MMP-13 gene expression in SW-1353 chondrosarcoma cells. Other investigators have demonstrated similar effects in chondrocytes treated with rosiglitazone, a ligand for peroxisome proliferator-activated receptor-gamma (PPAR gamma), for which RXR is an obligate dimerization partner. The goals of this study were to evaluate the inhibition of IL-1-beta-induced expression of MMP1 and MMP-13 by combinatorial treatment with RXR and PPAR gamma ligands and to investigate the molecular mechanisms of this inhibition.Methods We used real-time reverse transcription-polymerase chain reaction to measure LG268-and rosiglitazone-mediated inhibition of MMP gene transcription in IL-1-beta-treated SW-1353 chondrosarcoma cells. An in vitro collagen destruction assay was a functional readout of MMP collagenolytic activity. Luciferase reporter assays tested the function of a putative regulatory element in the promoters of MMP-1 and MMP-13, and chromatin immunoprecipitation (ChIP) assays detected PPAR gamma and changes in histone acetylation at this site. Posttranslational modification of RXR and PPAR gamma by small ubiquitinlike modifier (SUMO) was assayed with immunoprecipitation and Western blot.Results Rosiglitazone inhibited MMP-1 and MMP-13 expression in IL-1-beta-treated SW-1353 cells at the mRNA and heterogeneous nuclear RNA levels and blunted IL-1-beta-induced collagen destruction in vitro. Combining LG268 and rosiglitazone had an additive inhibitory effect on MMP-1 and MMP-13 transcription and collagenolysis. IL-1-beta-inhibited luciferase expression in the MMP reporter assay, but rosiglitazone and LG268 had no effect. ChIP indicated that treatment with IL-1-beta, but not LG268 and rosiglitazone, increased PPAR gamma at the proximal promoters of both MMPs. Finally, rosiglitazone or LG268 induced 'cross-SUMOylation' of both the target receptor and its binding partner, and IL-1-beta-alone had no effect on SUMOylation of RXR and PPAR gamma but antagonized the ligand-induced SUMOylation of both receptors.Conclusions The PPAR gamma and RXR ligands rosiglitazone and LG268 may act through similar mechanisms, inhibiting MMP-1 and MMP-13 transcription. Combinatorial treatment activates each partner of the RXR: PPAR gamma heterodimer and inhibits IL-1-beta-induced expression of MMP-1 and MMP-13 more effectively than either compound alone. We conclude that the efficacy of combined treatment with lower doses of each drug may minimize potential side effects of treatment with these compounds.