天津医药

天津医药 ›› 2015, Vol. 43 ›› Issue (6): 587-590.doi: 10.11958/j.issn.0253-9896.2015.06.003

• 细胞与分子生物学 • 上一篇    下一篇

β-蜕皮甾酮对脂多糖诱导兔软骨细胞损伤的保护作用

王刚涛,张旭辉,张卫东,夏磊   

  1. 解放军第371中心医院关节外科

  • 收稿日期:2014-10-11 修回日期:2015-01-14 出版日期:2015-06-15 发布日期:2015-06-10
  • 通讯作者: 王刚涛 E-mail:wanggangtao318@163.com

Protective effect of ecdysterone on rabbits chondrocytes that is injured by lipopolysaccharide

WANG Gangtao, ZHANG Xuhui, ZHANG Weidong, XIA Lei   

  1. Department of Joint surgery, the 371th Center Hospital of The PLA

  • Received:2014-10-11 Revised:2015-01-14 Published:2015-06-15 Online:2015-06-10

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摘要： 目的 探讨蜕皮甾酮 （EDS） 对脂多糖 （LPS） 诱导的兔软骨细胞损伤的保护作用及其机制。方法 体外分离、 培养兔关节软骨细胞， 随机分为对照组、 LPS 诱导损伤组（LPS 组）， 蜕皮甾酮干预组(LPS+EDS 组)。MTT 法和流式细胞术分别检测各组细胞增殖率及细胞凋亡率； RT-PCR 和 Western blot 检测软骨细胞中诱导型一氧化氮合酶（iNOS）表达； 硝酸还原酶法和 ELISA 法分别检测各组 NO 及白细胞介素（IL） -1β含量。结果 与对照组相比， LPS组细胞增殖率降低， 凋亡率升高， iNOS mRNA 和蛋白表达量以及 NO 和 IL-1β含量增高(均 P < 0.05)。LPS+EDS 组较LPS 组细胞增殖率升高， 凋亡率降低， iNOS mRNA 和蛋白表达量及 NO 和 IL-1β的含量降低（均 P < 0.05）。结论蜕皮甾酮对 LPS 诱导的兔软骨细胞损伤具有保护作用， 其保护作用可能与抑制 iNOS 表达有关

关键词: 蜕皮甾酮, 软骨细胞, 脂多糖类, 细胞增殖, 细胞凋亡, 白细胞介素 1β, 诱导型一氧化氮合酶

Abstract: Objective To study the effect of ecdysterone (EDS) on rabbits chondrocytes that is injuried by lipopolysaccharide (LPS). Methods Aricular chondrocytes were isolated from rabbits and randomly divided into three groups: control group; chondrocytes with LPS induced injury (LPS group); injury chondrocytes treated with EDS (LPS+EDS group). The cell proliferation and cell apoptosis of chondrocytes were determined by MTT method and flow cytometry assay respectively. The mRNA and protein expression levels of inducible nitric oxide synthase (iNOS) were detected by RT-PCR and western blot.In addition, the content of NO and IL-1β were measured by nitric acid reductase assay and enzyme-linked immunosorbent assay (ELISA) respectively. Results Attenuated proliferation, increased cell apoptosis, iNOS, NO and IL-1β were seen in LPS group , but all these changes were significantly reversed by EDS treatment (P < 0.05). Conclusion Ecdysterone exhibited a protective effect on LPS induced rabbits chondrocytes injury through inhibiting the expression of iNOS.

Key words: Ecdysterone, chondrocytes, lipopolysaccharides, cell proliferation, apoptosis, interleukin-1beta, iNOS