南方医科大学学报 ›› 2013, Vol. 33 ›› Issue (02): 225-.

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硫酸软骨素纳米硒可抑制T-2毒素诱导的大骨节病软骨细胞凋亡

韩晶,郭雄,吴翠艳,李春燕,何淑兰,段琛,宁玉洁   

  • 出版日期:2013-02-20 发布日期:2013-02-20

Nano-Se-chondroitin sulfate inhibits T-2 toxin-induced apoptosis of cultured chondrocytes from patients with Kashin-Beck disease

  • Online:2013-02-20 Published:2013-02-20

摘要: 目的确定硫酸软骨素纳米硒对T-2毒素干预体外大骨节病软骨细胞生长的影响。方法合成并表征硫酸软骨素纳米硒粒
子,依据《大骨节病临床诊断标准》(WS/T 207-2010),选择Ⅱ/Ⅲ度KBD患者6例关节软骨进行体外分离、培养。分别给予硫酸
软骨素纳米硒联合T-2毒素进行干预,采用MTT、HE染色和流式细胞仪观察细胞生长和凋亡的变化。结果合成的硫酸软骨素
纳米硒中硒的含量为10.1%,可在蒸馏水中自组装成粒径为30~200 nm纳米粒子,红外图谱提示纳米硒与硫酸软骨素可能以共
价键的方式结合;硫酸软骨素纳米硒在浓度为50~200 ng/ml可有效抑制20 ng/ml T-2毒素诱导大骨节病软骨细胞的凋亡作用,
降低T-2毒素诱导软骨细胞的早期凋亡率[(8.64±1.57)%,P<0.05]。结论硫酸软骨素纳米硒具有抗T-2毒素诱导软骨细胞凋亡
的作用,可作为一种潜在的治疗大骨节病的药物制剂。

Abstract: Objective To observe the effect of nano-Se-chondroitin sulfate on the growth and apoptosis of chondrocytes from
patients with Kashin-Beck disease (KBD) exposed to T-2 toxin in vitro. Methods Samples of the articular cartilage were
obtained from 6 patients with grade II/III KBD diagnosed in line with the National Clinical Diagnostic Criteria of KBD (WS/T
207-2010) for chondrocyte separation and culture in vitro. The separated chondrocytes were treated with synthesized
nano-Se-chondroitin sulfate particles and T-2 toxin, alone or in combination, and the cell growth and apoptosis were observed
using MTT assay, HE staining and flow cytometry. Results The synthesized nano-Se-chondroitin sulfate, with a selenium
entrapment ratio of 10.1%, spontaneously formed nanoparticles in distilled water with sizes ranging from 30 to 200 nm.
Fourier-transform infrared spectroscopy suggested a possible covalent bond that bound Nano-Se and chondroitin sulfate.
Within the concentration range of 50-200 ng/ml, nano-Se-chondroitin sulfate significantly inhibited T-2 toxin-induced
apoptosis of the cultured chondrocytes and reduced the early apoptosis rate to (8.64±1.57)% (P<0.05). Conclusion Nano-Sechondroitin
sulfate can inhibit T-2 toxin-induced apoptosis of cultured chondrocytes from KBD patients in vitro, and serves as
a promising candidate therapeutic agent for KBD.