雪腐镰刀菌烯醇对培养软骨细胞蛋白聚糖合成的影响

Influence of nivalenol on aggrecan synthesis of cultured chondrocytes

背景：雪腐镰刀菌烯醇是真菌毒素之一,它可能与大骨节病的发生具有一定的相关性。目的：验证雪腐镰刀菌烯醇对软骨细胞蛋白聚糖合成的影响。方法：在体外单层培养的人胚软骨细胞中加入不同质量浓度（0.025,0.05,0.1,0.2,0.4,0.8,1.6mg/L）的雪腐镰刀菌烯醇毒素,作用1~5d后收集软骨细胞,用MTT法检测软骨细胞存活率;用紫外分光光度法检测细胞DNA含量,RT-PCR方法检测蛋白聚糖mRNA表达;用Westernblot法检测人软骨细胞蛋白聚糖表达。结果与结论：0.025mg/L雪腐镰刀菌烯醇毒素可刺激软骨细胞生长,其刺激作用呈先升高后降低的趋势。0.05~0.1mg/L毒素作用早期的细胞存活率增加,随后细胞生长被毒素抑制。当毒素质量浓度升高至0.2mg/L以上时,随着雪腐镰刀菌烯醇毒素质量浓度的增加及作用时间延长,细胞存活率明显下降。0.1~0.5mg/L雪腐镰刀菌烯醇毒素可以抑制软骨细胞蛋白聚糖的合成及其mRNA表达。结果表明雪腐镰刀菌烯醇毒素对软骨细胞蛋白聚糖合成有明显的抑制作用,并以剂量和时间依赖性方式影响细胞的增殖。

BACKGROUND： Nivalenol （NIV） toxin is one of the mycotoxins, which is supposed to probably contribute to the classical features of Kashin-beck disease. OBJECTIVE： To investigate the influence of NIV on the expression of aggrecan in the cultured chondrocytes. METHODS： Human chondrocytes cultured in vitro were treated with NIV at different concentrations for varied time periods （1- 5 days）, and the cell viability was measured by MTT assay. DNA content was determined by UV Spectrophotometry. Aggrecan mRNA expression was studied by semiquantitative RT-PCR. The protein expression of aggrecan was examined by using Western blot analysis. RESULTS AND CONCLUSION： NIV at a dose of 0.025 mg/L could stimulate chondrocytes growth in a tendency of increasing followed by decreasing. The chondrocytes viability was initially increased when treated with NIV, which would be suppressed by toxin when treated with NIV at a dose of 0.05-0.1 mg/L. When treated with more than 0.2 mg/L NIV, the chondrocytes viability was obviously decreased with concentration increasing and time prolonging. The NIV with concentration of 0.1-0.5 mg/L inhibited aggrecan synthesis and reduced its mRNA expression. The results demonstrated that NIV significantly decrease aggrecan synthesis in the chondrocytes and reduced its mRNA expression. NIV could inhibit chondrocyte proliferation in a dose-and time-dependent manner.