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

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

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。

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 MTF 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-Se- chondroitin 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.