MMP13 is a critical target gene during the progression of osteoarthritis.
MMP13 is a critical target gene during the progression of osteoarthritis.
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DOI：
10.1186/ar4133
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发表时间：
2013-01-08
影响因子：
4.9
通讯作者：
Chen D
中科院分区：
文献类型：
--
作者：
Wang M;Sampson ER;Jin H;Li J;Ke QH;Im HJ;Chen D
骨关节炎（OA）是一种退行性关节疾病，影响了大量人群。这种高度流行的疾病的机制尚未完全了解。目前还没有有效的疾病缓解治疗OA。本研究有两个目的：1）研究MMP 13在OA发生中的作用; 2）评价MMP 13抑制剂CL 82198作为预防OA进展的药物治疗的疗效。为了研究内源性Mmp 13基因在OA发展中的作用，将他莫昔芬给予两周龄的Col 2CreER; Mmp 13 fx/fx（Mmp 13 Col 2 ER）和Cre阴性对照小鼠5天。当小鼠为10周龄时，通过踝关节韧带损伤（MLI）诱导OA，并在手术后4、8、12或16周收获MLI或假手术关节。为了评估CL 82198的功效，对10周龄野生型小鼠进行MLI手术。在手术后立即开始每天向小鼠施用CL 82198或盐水，持续长达16周。通过软骨分级、组织学/组织形态计量学、免疫组织化学（IHC）和末端脱氧核苷酸转移酶dUTP缺口末端标记（TUNEL）染色评价从两个实验中收集的关节组织。通过ELISA证实CL 82198体外抑制MMP 13活性的能力。术后8、12和16周，Mmp 13 Col 2 ER小鼠的OA进展减慢。由盲法观察者进行的软骨分级证实，与Cre阴性小鼠相比，Mmp 13 Col 2 ER小鼠在8、12和16周时的关节软骨变性减少。组织形态计量学分析表明，Mmp 13 Col 2 ER小鼠在术后12周和16周与对照小鼠相比具有更高的关节软骨面积和厚度。免疫组化结果显示，Mmp 13 Col 2 ER小鼠中II型胶原和蛋白多糖的表达更高。软骨细胞凋亡，通过TUNEL染色测定，在对照组小鼠相比，Mmp 13 Col 2 ER小鼠。CL 82198抑制来自溶剂（> 85%）或骨形态发生蛋白2（BMP 2）处理（> 90%）的原代小鼠胸骨软骨细胞的条件培养基中的MMP 13活性。与生理盐水处理相比，腹腔注射CL 82198可减缓MLI诱导的OA进展，增加II型胶原和蛋白聚糖水平，并抑制软骨细胞凋亡，分别通过OA分级、组织学、组织形态学、IHC和TUNEL染色确定。Mmp 13对OA进展至关重要，MMP 13的药理学抑制是在损伤诱导的膝关节OA小鼠模型中减缓关节软骨损失的有效策略。
Osteoarthritis (OA) is a degenerative joint disease affecting a large population of people. The mechanism of this highly prevalent disease is not fully understood. Currently there is no effective disease-modifying treatment for OA. The purpose of this study was two-fold: 1) to investigate the role of MMP13 in the development of OA; and 2) to evaluate the efficacy of the MMP13 inhibitor CL82198 as a pharmacologic treatment for preventing OA progression. To investigate the role of the endogenous Mmp13 gene in OA development, tamoxifen was administered to two-week-old Col2CreER;Mmp13fx/fx (Mmp13Col2ER) and Cre-negative control mice for five days. OA was induced by meniscal-ligamentous injury (MLI) when the mice were 10 weeks old and MLI or sham-operated joints were harvested 4, 8, 12, or 16 weeks after surgery. To evaluate the efficacy of CL82198, MLI surgery was performed on 10-week-old wild type mice. CL82198 or saline was administered to the mice daily beginning immediately after the surgery for up to 16 weeks. The joint tissues collected from both experiments were evaluated by cartilage grading, histology/histomorphometry, immunohistochemistry (IHC), and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining. The ability of CL82198 to inhibit MMP13 activity in vitro was confirmed by ELISA. The OA progression was decelerated in Mmp13Col2ER mice 8, 12, and 16 weeks post-surgery. Cartilage grading by blinded observers confirmed decreased articular cartilage degeneration in Mmp13Col2ER mice at 8, 12 and 16 weeks compared to Cre-negative mice. Histomorphometric analysis demonstrated that Mmp13Col2ER mice had a higher articular cartilage area and thickness at 12 and 16 weeks post-surgery compared to the control mice. Results of IHC revealed greater type II collagen and proteoglycan expression in Mmp13Col2ER mice. Chondrocyte apoptosis, as determined by TUNEL staining, was higher in control mice compared to Mmp13Col2ER mice. CL82198 inhibited MMP13 activity in conditioned media from vehicle (> 85%) or bone morphogenetic protein 2 (BMP2)-treated (> 90%) primary murine sternal chondrocytes. Intraperitoneal injection of CL82198 decelerated MLI-induced OA progression, increased type II collagen and proteoglycan levels, and inhibited chondrocyte apoptosis compared to saline treatment as determined by OA grading, histology, histomorphometry, IHC, and TUNEL staining, respectively. Mmp13 is critical for OA progression and pharmacologic inhibition of MMP13 is an effective strategy to decelerate articular cartilage loss in a murine model of injury-induced knee OA.
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DOI：
10.1186/ar401
发表时间：
2002
期刊：
Arthritis research
影响因子：
--
作者：
Vincenti MP;Brinckerhoff CE
通讯作者：
Brinckerhoff CE
影响因子：
7
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Wang M;Jin H;Tang D;Huang S;Zuscik MJ;Chen D
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影响因子：
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作者：
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通讯作者：
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影响因子：
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作者：
Clements, KM;Price, JS;Mason, RM
通讯作者：
Mason, RM
影响因子：
15
作者：
Chen, JM;Nelson, FC;Powers, R
通讯作者：
Powers, R