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糖代谢紊乱对大骨节病软骨细胞功能的影响

The Effect of Disordered Glycometabolism of Kashin-Beck Disease on the Function of Chondrocytes

  • 摘要: 目的观察大骨节病糖代谢紊乱对软骨细胞外基质成分胶原和蛋白多糖、细胞凋亡以及氧化应激物质产生的作用,揭示糖代谢异常对大骨节病软骨细胞功能的影响。方法正常培养基和高浓度葡萄糖(高糖)培养基分别体外培养的大骨节病软骨细胞和正常软骨细胞,进行细胞增殖和形态学观察,检测软骨细胞培养基中葡萄糖的浓度,采用实时荧光定量PCR(qRT-PCR)、甲苯胺蓝染色等方法检测两组软骨细胞Ⅱ型胶原和蛋白多糖的表达,流式细胞术检测细胞凋亡和活性氧积累。结果大骨节病软骨细胞生长速度和增殖能力低于正常软骨细胞。高糖环境下正常软骨细胞和大骨节病软骨细胞对葡萄糖的代谢量基本相同(P>0.05)。高糖造成的糖代谢紊乱使得大骨节病软骨细胞Ⅱ型胶原和蛋白多糖的表达量均降低(P<0.05)。同时,高糖培养下,大骨节病软骨细胞的凋亡率和细胞内活性氧的含量均增加(P<0.05)。结论紊乱的糖代谢降低大骨节病软骨细胞Ⅱ型胶原和蛋白多糖的表达,增加细胞凋亡和氧化应激反应,从而影响软骨细胞的功能。

     

    Abstract: Objective To reveal the effect of disordered glycometabolism in Kashin-Beck disease (KBD) chondrocytes, we compared changes in expressions of extracellular matrix components (collagen and aggrecan), apoptosis and oxidative stress under the condition of different concentrations of glucose. Methods The damage of KBD chondrocytes and normal chondrocytes under high glucose culture was measured in compared with cells under normal culture, that included the changes of proliferation and morphology; the concentrations of glucose in culture medium during the process of chondrocytes culture; the expressions of type Ⅱ collagen and aggrecan detected by quantitative real-time polymerase chain reaction (qRT-PCR) and Toluidine blue staining; cell apoptosis and reactive oxygen species (ROS) content detected by flow cytometry and fluorescence staining. Results The growth and proliferation of KBD chondrocytes were inferior to normal chondrocytes. The glucose uptake of KBD chondrocytes and normal chondrocytes under high glucose culture were basically the same (P>0.05). Disordered glycometabolism caused by high glucose decreased the expression of type Ⅱ collagen and aggrecan in KBD chondrocytes (P<0.05), meanwhile, increased apoptosis and cellular ROS generation of cultured chondrocytes (P<0.05). Conclusion The disordered glycometabolism can affect the function of KBD chondrocytes through reducing the expression of type Ⅱ collagen and aggrecan and increasing the apoptosis and the oxidative stress.

     

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