CAJ | 학술논문

目的比较纳米硒(Nano-Se)、亚硒酸钠(Na2SeO3)对体外培养的大骨节病关节软骨细胞凋亡和活性氧的影响,为大骨节病的药物治疗提供科学依据.方法依据《大骨节病临床诊断》标准(WS/T207-2010),收集8例进行Ⅰ、Ⅱ度大骨节病膝关节置换病例的关节软骨.分离软骨组织,得到软骨细胞.分别给予0、25、50、100、200、300、400、500 μg/L Nano-Se和Na2SeO3培养液培养5d,采用四氮唑蓝(MTT)法检测软骨细胞存活率,以Nano-Se和Na2SeO3干预后的最高存活率对应的浓度与时间作为后续实验条件.取软骨细胞,分为对照组、Na2SeO3组、Nano-Se组,每组8例,以上述得到的最佳实验浓度和时间进行培养,采用流式细胞术测定法测定软骨细胞凋亡率及活性氧荧光强度.结果用Nano-Se、Na2SeO3干预后,软骨细胞最高存活率对应的最佳浓度分别为100、400 μg/L,最佳干预时间均为3d.Nano-Se组、Na2SeO3组的总凋亡率[(4.67±0.89)％、(7.07±0.25)％]、晚期凋亡率[(1.51±0.48)％、(4.37±0.37)％]均低于对照组[(9.95±0.38)％、(6.93±0.42)％,P均＜0.01],Nano-Se组的总凋亡率、晚期凋亡率均低于Na2SeO3组(P均＜0.01).对照组、Na2SeO3组、Nano-Se组的早期凋亡率[(3.02±0.41)％、(2.7±0.46)％、(3.16±0.56)％]组间比较,差异无统计学意义(F=2.11,P=0.35).Nano-Se组、Na2SeO3组软骨细胞活性氧荧光强度[(87.13±6.60)％、(56.04±4.81)％]均低于对照组[(125.17±16.60)％,P＜ 0.01],Nano-Se组低于Na2SeO3组(P＜0.01).结论适宜浓度纳米硒较亚硒酸钠能更有效地缓解大骨节病软骨细胞的氧化应激水平,降低细胞凋亡,为深入研究大骨节病药物治疗提供了新的科学依据.
목적 비교납미서(Nano-Se)、아서산납(Na2SeO3)대체외배양적대골절병관절연골세포조망화활성양적영향,위대골절병적약물치료제공과학의거.방법 의거《대골절병림상진단》표준(WS/T207-2010),수집8례진행Ⅰ、Ⅱ도대골절병슬관절치환병례적관절연골.분리연골조직,득도연골세포.분별급여0、25、50、100、200、300、400、500 μg/L Nano-Se화Na2SeO3배양액배양5d,채용사담서람(MTT)법검측연골세포존활솔,이Nano-Se화Na2SeO3간예후적최고존활솔대응적농도여시간작위후속실험조건.취연골세포,분위대조조、Na2SeO3조、Nano-Se조,매조8례,이상술득도적최가실험농도화시간진행배양,채용류식세포술측정법측정연골세포조망솔급활성양형광강도.결과 용Nano-Se、Na2SeO3간예후,연골세포최고존활솔대응적최가농도분별위100、400 μg/L,최가간예시간균위3d.Nano-Se조、Na2SeO3조적총조망솔[(4.67±0.89)％、(7.07±0.25)％]、만기조망솔[(1.51±0.48)％、(4.37±0.37)％]균저우대조조[(9.95±0.38)％、(6.93±0.42)％,P균＜0.01],Nano-Se조적총조망솔、만기조망솔균저우Na2SeO3조(P균＜0.01).대조조、Na2SeO3조、Nano-Se조적조기조망솔[(3.02±0.41)％、(2.7±0.46)％、(3.16±0.56)％]조간비교,차이무통계학의의(F=2.11,P=0.35).Nano-Se조、Na2SeO3조연골세포활성양형광강도[(87.13±6.60)％、(56.04±4.81)％]균저우대조조[(125.17±16.60)％,P＜ 0.01],Nano-Se조저우Na2SeO3조(P＜0.01).결론 괄의농도납미서교아서산납능경유효지완해대골절병연골세포적양화응격수평,강저세포조망,위심입연구대골절병약물치료제공료신적과학의거.
Objective To compare the effects of selenium nanoparticles (Nano-Se) and sodium selenium (Na2SeO3) on apoptosis and reactive oxygen species (ROS) of articular chondrocytes from patients with Kashin-Beck Disease (KBD) in vitro,and provide a scientific basis for preventing KBD.Methods The subjects with KBD were diagnosed on National Clinical Diagnostic Criteria of KBD (WS/T207-2010),articular cartilage from 8 patients undertaken joint replacement operation were collected.In vitro,chondrocytes were treated with concentration of 0,25,50,100,200,300,400 and 500 μg/L of Nano-Se and Na2SeO3 for 5 d,respectively.Cell growth was detected by MTT assay,and the highest concentration and time corresponding to the highest survival rate of Nano-Se and Na2SeO3 were used in the following experiment.KBD chondrocytes were treated with Nano-Se and Na2SeO3,and divided into control group,Na2SeO3 group,Nano-Se group according to the randomized design.Each group had 8 cases.The cell apoptosis and ROS were detected by flow cytometry.Results The optimal intervention concentration of Nano-Se and Na2SeO3 was 100 and 400 μg/L,respectively.The optimal intervention time of NanoSe and Na2SeO3 both was 3 days.There was a significant decrease in the total and terminal apoptosis,ROS level of chondrocytes in Nano-Se group [(4.67 ± 0.89)％,(1.51 ± 0.48)％,(56.04 ± 4.81)％] and Na2SeO3 group [(7.07 ±0.25)％,(4.37 ± 0.37)％,(87.13 ± 6.60)％] compared with those of control group [(9.95 ± 0.38)％,(6.93 ± 0.42)％,(125.17 ± 16.60)％,all P ＜ 0.01].The difference of early apoptotic rate among control group,Na2SeO3 group,NanoSe group [(3.02 ± 0.41)％,(2.7 ± 0.46)％,(3.16 ± 0.56)％] was not statistically significant (F =2.11,P =0.35).Conclusion Appropriate concentration of Nano-Se can significantly decrease oxidative stress of KBD chondrocytes and inhibit apoptosis compared to Na2SeO3.