锌与金雀异黄素对小鼠胚胎肢芽的影响

THE EFFECT OF ZINC AND GENISTEIN ON MOUSE LIMB BUD IN VITRO

[目的]研究锌与金雀异黄素对软骨发育分化的影响。[方法]利用12d小鼠胚胎肢芽体外培养方法,在培养基中分别加入10-7～10-5mol/L不同含量的金雀异黄素以及在加入10-5mol/L金雀异黄素的同时加ZnSO450μmol/L和70μmol/L,并设对照组,培养3d后,收获肢芽用原位杂交的方法测量肢芽软骨细胞II型胶原mRNA及BMP-2mR-NA的表达水平以及IL-6的表达水平。[结果]体外培养3d的肢芽在金雀异黄素(10-5mol/L)组与金雀异黄素(10-5mol/L)加ZnSo4组的II型胶原mRNA及BMP-2mRNA阳性表达密度与积分光密度显著高于对照组;体外培养3d的IL-6表达水平在金雀异黄素(10-5mol/L)组与金雀异黄素(10-5mol/L)加ZnSo4组显著低于对照组。[结论]锌与金雀异黄素在一定浓度下能协同促进胚胎肢芽软骨细胞BMP-2mRNA及II型胶原mRNA的表达,以及降低IL-6的表达水平,促进发育中软骨面积的增加,具有刺激胚胎软骨发育分化的作用。

[Objective] To study the effect of zinc and genistein on the cartilage development of mouse fetal limb bud in vitro. [Methods] The fetal limb buds of 12-day gestational period were cultured in the rotating apparatus in vitro for 3 days. Genistein from 10^-5 mol/L to 10^-5 mol/L and Zinc （50 μmol/L, 70 μmol/L）were added to the modium. The expression of type 2 collagen mRNA, BMP-2 mRNA and IL-6 were determined by in situ hybridization. [ Results] After culturing for 3 days, the density and IOD of positive expression of type 2 collagen mRNA and BMP-2 mRNA of chonddroeyte in genistein 10^-5 mol/L and Zinc 50 μmol/L-70 μmol/L group were significantly higher than that of the eontal. IL-6 in gemstein 10-^5 mol/L and zinc 50 μmol/L-70 μmol/L group were significantly lower than that of the control. [ Conclusions ] Genistein and zinc increase the expression of BMP-2 mRNA and type 2 collagen mRNA and IL-6 of the limb bud chOndroeyte, enlarge the area of the cartilage, promotes the development and differentiation of the cartilage of limb bud.