NIV毒素和硒对软骨细胞CD44代谢的影响

摘要

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,加硒组加人硒浓度为0.1 mg/L,雪腐镰刀菌烯醇毒素(NIV)组加人NIV毒素使终质量浓度为0.1mg/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(aCD44)水平.结果 NIV组、联合组软骨细胞CD,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入水平最高,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 patlogenesis of Kashin-Beck Disease further. Methods Chondrocytes were isolated from a 4-month-old human fetal cartilage and cuhred in vitro, and then were divided into 4 groups. The control group was not treated, Se group was given Se of 0.1 mg/L, nivalenol (NIV) group was given NIV of 0.1 mg/L, and joined group was given Se and NIV with aboved dose. the expression of CD44 mRNA was measured by RT-PCR; the expression of CD44 protein on the chondrocyte membrane was determined by flow cytometer; the level of soluble CD44 ( sCD44 )in cultured medium was measured by enzyme-linked immunosorbent assay (ELISA). Results The expression of CD44 mRNA and CD44 protein in NIV group and joined group were all significantly higher than those in control group and Se group ( P＜0.05). Compared with NIV group, the expression of CD44 decreased in joined group, but with no significant difference.The content of sCD44 in medium of NIV group was highest than that in other groups, secondly was the joined group, and that of control group was the lowest. Conclusions NIV toxin and Se can effect the metabolism of CD44, which may eventually lead to the metabolic disorder of the cartilage matrix components.