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目的进一步探讨大骨节病(KBD)的发病机制。方法取4月龄引产胎儿软骨细胞原代分离、培养并随机分为四组。对照组不处理,加硒组加入硒浓度为0.1 mg/L,雪腐镰刀菌烯醇毒素(NIV)组加入NIV毒素使终质量浓度为0.1 mg/L,联合组加硒和NIV毒素,浓度同上。RT-PCR法检测各组CD44 mRNA在软骨细胞中的表达;流式细胞仪检测各组软骨细胞表面CD44蛋白的表达;酶联免疫吸附法检测细胞培养液中可溶性CD44(sCD44)水平。结果 NIV组、联合组软骨细胞CD44 mRNA及蛋白表达水平均明显高于对照组和加硒组(P均<0.05)。与NIV组比较,联合组CD44表达有所降低,但不明显。NIV组细胞培养液中溶解的sCD44水平最高,其次为联合组,对照组水平最低。结论 NIV毒素和硒能影响软骨细胞CD44的代谢,这可能是造成软骨细胞损伤的机制。
Objective To explore the pathogenesis of Kashin-Beck disease (KBD). Methods Primary fetal chondrocytes from 4-month old abortion were isolated, cultured and randomly divided into four groups. In the control group, selenium concentration was 0.1 mg / L, NIV toxin was added to make the final concentration 0.1 mg / L, selenium and NIV toxin in the combination group, Ibid. The expression of CD44 mRNA in chondrocytes of each group was detected by RT-PCR. The expression of CD44 on the chondrocytes was detected by flow cytometry. The level of soluble CD44 (sCD44) was detected by enzyme-linked immunosorbent assay (ELISA). Results The levels of CD44 mRNA and protein in chondrocytes of NIV group and combination group were significantly higher than those of control group and selenium group (all P <0.05). Compared with the NIV group, the expression of CD44 in the combined group was decreased, but not obvious. The highest level of sCD44 was dissolved in the cell culture medium of NIV group, followed by the combination group and the lowest level in the control group. Conclusion NIV toxin and selenium can affect the metabolism of CD44 in chondrocytes, which may be the mechanism of chondrocyte injury.