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首页> 外文期刊>Arthritis & Rheumatism >Cytokine-induced increases in ADAMTS-4 messenger RNA expression do not lead to increased aggrecanase activity in ADAMTS-5–deficient mice
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Cytokine-induced increases in ADAMTS-4 messenger RNA expression do not lead to increased aggrecanase activity in ADAMTS-5–deficient mice

机译：细胞因子诱导的ADAMTS-4信使RNA表达的增加不会导致ADAMTS-5缺陷小鼠中聚集蛋白聚糖酶活性的增加

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ObjectiveTo compare the regulation of aggrecanase messenger RNA (mRNA) and enzyme activity by proinflammatory cytokines in primary mouse chondrocytes.MethodsPrimary chondrocytes were isolated from knee epiphyses of 6–8-day-old mice and cultured as monolayers. The cells were incubated with tumor necrosis factor (TNF), oncostatin M (OSM), or interleukin-6 (IL-6)/soluble IL-6 receptor, and mRNA levels were measured by quantitative polymerase chain reaction at various time points. To measure aggrecanase activity, the cells were incubated with cytokine in the presence of exogenous aggrecan, and substrate cleavage was measured using antibodies to neoepitopes.ResultsExpression of both ADAMTS-4 and ADAMTS-5 mRNA was up-regulated by TNF and OSM. ADAMTS-5 mRNA expression was also up-regulated by IL-6. Treatment of wild-type mouse chondrocytes with each of the 3 cytokines increased cleavage of aggrecan at Glu373↓374Ala and Glu1670↓1671Gly; in chondrocytes lacking ADAMTS-5 activity, there was negligible cleavage at either site despite increased expression of ADAMTS-4 mRNA in the presence of TNF or OSM. None of the cytokines substantially altered mRNA expression of ADAMTS-1 or ADAMTS-9.ConclusionDespite substantial increases in the expression of ADAMTS-4 mRNA induced by TNF and OSM, these cytokines induced little if any increase in aggrecanolysis in ADAMTS-5–deficient mouse chondrocytes. Our data show a poor correlation between the level of cytokine-induced ADAMTS-4 mRNA expression and the level of aggrecan-degrading activity in cultured chondrocytes.
机译：目的比较促炎细胞因子对小鼠原代软骨细胞中软骨聚集蛋白聚糖信使RNA（mRNA）的调节和酶活性。方法从6-8日龄小鼠的膝骨ses中分离原代软骨细胞，并进行单层培养。将细胞与肿瘤坏死因子（TNF），制瘤素M（OSM）或白介素6（IL-6）/可溶性IL-6受体一起孵育，并通过定量聚合酶链反应在不同时间点测量mRNA水平。为了测量软骨聚集蛋白聚糖酶的活性，在存在外源性软骨聚集蛋白聚糖的情况下，将细胞与细胞因子一起孵育，并使用针对新表位的抗体测量底物的裂解。结果TNF和OSM均上调了ADAMTS-4和ADAMTS-5 mRNA的表达。 ADAMTS-5 mRNA表达也被IL-6上调。用三种细胞因子中的每一种处理野生型小鼠软骨细胞均会增加Glu 373 ↓ 374 Ala和Glu 1670 ↓ 1671 Gly;在缺乏ADAMTS-5活性的软骨细胞中，尽管在存在TNF或OSM的情况下ADAMTS-4 mRNA的表达增加，但在任一位点的切割均可以忽略不计。结论：尽管TNF和OSM诱导ADAMTS-4 mRNA表达显着增加，但这些细胞因子几乎没有诱导ADAMTS-5缺陷小鼠聚集蛋白水解的增加。软骨细胞。我们的数据显示细胞因子诱导的ADAMTS-4 mRNA表达水平与培养的软骨细胞中聚集蛋白聚糖降解活性水平之间的相关性较差。

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