An altered heparan sulfate structure in the articular cartilage protects against osteoarthritis.
An altered heparan sulfate structure in the articular cartilage protects against osteoarthritis.
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DOI：
10.1016/j.joca.2020.04.002
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发表时间：
2020-07
影响因子：
7
通讯作者：
Vortkamp A
中科院分区：
文献类型：
--
作者：
Severmann AC;Jochmann K;Feller K;Bachvarova V;Piombo V;Stange R;Holzer T;Brachvogel B;Esko J;Pap T;Hoffmann D;Vortkamp A
骨关节炎（OA）是由蛋白酶、细胞因子和其他分泌蛋白的不平衡活性引起的关节软骨的进行性退行性疾病。由于硫酸乙酰肝素（HS）决定了许多细胞外因子的活性，我们研究了其在OA进展中的作用。为了分析HS水平的作用，在软骨细胞克隆中携带Ext 1功能丧失等位基因的转基因小鼠（Col 2-rtTA-Cre; Ext 1 e2 fl/e2 fl）中，通过前交叉韧带横断（ACLT）诱导OA。为了研究HS硫酸化模式的影响，在具有磺基转移酶Ndst 1的杂合（Ndst 1 +/−）或软骨细胞特异性（Col 2-Cre; Ndst 1fl/fl）功能丧失等位基因的小鼠中手术诱导OA。使用OARSI评分系统评价OA进展。为了研究软骨降解蛋白酶的表达和活性，通过qRT-PCR、蛋白质印迹和明胶酶谱分析Ndst 1 +/−突变体的股骨头外植体。所有研究的小鼠品系均显示OA评分降低（Col 2-rtTA-Cre; Ext 1 e2 fl/e2 fl：0.83; 95%HDI 0.72-0.96; Ndst 1 +/−：0.83，95%HDI 0.74-0.9; Col 2-Cre; Ndst 1fl/fl：0.87，95%HDI 0.76-1）。使用Ndst 1动物的软骨外植体培养物，我们在野生型样品中检测到更高量的聚集蛋白聚糖降解产物（NITEGE 4.24倍，95%HDI 1.05-18.55; VDIPEN 1.54倍，95%HDI 1.54-2.34）。因此，明胶酶谱显示RA处理后突变体样品中Mmp 2活性较低（0.77倍，95%HDI：0.60-0.96）。由于主要蛋白酶及其抑制剂的表达没有改变，HS似乎通过影响蛋白酶活性来调节软骨退变。降低HS含量或降低硫酸化水平通过调节蛋白酶活性而不是表达来防止OA进展。
Osteoarthritis (OA) is a progressive degenerative disease of the articular cartilage caused by an unbalanced activity of proteases, cytokines and other secreted proteins. Since heparan sulfate (HS) determines the activity of many extracellular factors, we investigated its role in OA progression. To analyze the role of the HS level, OA was induced by anterior cruciate ligament transection (ACLT) in transgenic mice carrying a loss-of-function allele of Ext1 in clones of chondrocytes (Col2-rtTA-Cre;Ext1e2fl/e2fl). To study the impact of the HS sulfation pattern, OA was surgically induced in mice with a heterozygous (Ndst1+/−) or chondrocyte-specific (Col2-Cre;Ndst1fl/fl) loss-of-function allele of the sulfotransferase Ndst1. OA progression was evaluated using the OARSI scoring system. To investigate expression and activity of cartilage degrading proteases, femoral head explants of Ndst1+/− mutants were analyzed by qRT-PCR, Western Blot and gelatin zymography. All investigated mouse strains showed reduced OA scores (Col2-rtTA-Cre;Ext1e2fl/e2fl: 0.83; 95% HDI 0.72–0.96; Ndst1+/−: 0.83, 95% HDI 0.74–0.9; Col2-Cre;Ndst1fl/fl: 0.87, 95% HDI 0.76–1). Using cartilage explant cultures of Ndst1 animals, we detected higher amounts of aggrecan degradation products in wildtype samples (NITEGE 4.24-fold, 95% HDI 1.05–18.55; VDIPEN 1.54-fold, 95% HDI 1.54–2.34). Accordingly, gelatin zymography revealed lower Mmp2 activity in mutant samples upon RA-treatment (0.77-fold, 95% HDI: 0.60–0.96). As expression of major proteases and their inhibitors was not altered, HS seems to regulate cartilage degeneration by affecting protease activity. A decreased HS content or a reduced sulfation level protect against OA progression by regulating protease activity rather than expression.
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影响因子：
2
作者：
KOCH, AL
通讯作者：
KOCH, AL
影响因子：
2
作者：
Otsuki, Shuhei;Murakami, Tomohiko;Neo, Masashi
通讯作者：
Neo, Masashi
DOI：
10.1073/pnas.0910875107
发表时间：
2010-02-02
影响因子：
11.1
作者：
Jones, Kevin B.;Piombo, Virginia;Sheffield, Val C.
通讯作者：
Sheffield, Val C.
影响因子：
15.9
作者：
Fosang, AJ;Last, K;Maciewicz, RA
通讯作者：
Maciewicz, RA
影响因子：
7.8
作者：
Holzer, Tatjana;Probst, Kristina;Brachvogel, Bent
通讯作者：
Brachvogel, Bent