目的:研究脱氢表雄酮(DHEA)对白细胞介素(IL-1β)诱导的人骨关节炎软骨细胞退变的影响。方法:分离人骨关节炎软骨细胞传代培养并加入IL-1β,分别将50,25和12.5μmol/L的DHEA作用于该软骨细胞。其后检测细胞增殖、上清液糖胺聚糖(GAG)和NO含量、细胞凋亡以及质金属蛋白酶-1及其组织抑制剂-1(MMP-1和TIMP-1)含量。结果:50,25和12.5μmol/L的DHEA均能促进软骨细胞增殖和GAG合成(均P〈0.01),抑制IL-1β诱导的软骨细胞凋亡(均P〈0.01)以及MMP-1合成(均P〈0.01);50和25μmol/L的DHEA可抑制IL-1β诱导的软骨细胞NO形成(均P〈0.01),还可抑制软骨细胞自身的凋亡(均P〈0.01)和MMP-1合成(均P〈0.05),其效应呈浓度依赖性;50μmol/L的DHEA还可改善IL-1β对骨关节炎软骨细胞TIMP-1合成的抑制作用(P〈0.05)。结论:脱氢表雄酮能在一定程度上对抗IL-1β诱导的人骨关节炎软骨细胞退变。
Objective: To investigate the effects of dehydroepiandrosterone(DHEA) on in vitro human osteoarthritic chondrocytes induced by IL-1β.Methods: Chondrocytes isolated from human osteoarthritic knee cartilage were cultured and passaged.After monolayers were established,chondrocytes induced by IL-1β were treated with DHEA of 50,25 and 12.5 μmol/L respectively.The effects on chondrocytes were analyzed using 3-(4,5-dimethylthiazol-2yl)-5-(3-carboxymethoxy-phenyl)-2-(4-sulfophenyl)-2H-tetrazoliuminner salt(MTS) assay for chondrocyte proliferation and dimethylmethylene blue assay for the content of glycosaminoglycan(GAG).The content of nitric oxide(NO) in cultured supernate was measured by nitrate reductase method.Cell cycle and apoptosis of the chondrocytes were evaluated by flow cytometry.Protein contents of metalloproteinase-1(MMP-1) and tissue inhibitor of metalloproteinase-1(TIMP-1) were determined by Western blotting.Results: ① DHEA had no toxic effect on chondrocytes in all observed concentrations.②DHEA with a concentration of 50,25,or 12.5 μmol/L up-regulated chondrocyte proliferation(all P0.01) and GAG synthesis(all P0.01),inhibited chondrocyte apoptosis induced by IL-1β(all P0.01),and the MMP-1 synthesis of the osteoarthritic chondrocytes induced by IL-1β was also inhibited(all P0.01).③DHEA with a concentration of 50 μmol/L or 25 μmol/L inhibited NO formation induced by IL-1β(both P0.01).Simultaneously,the intrinsic apoptosis and MMP-1 synthesis of the osteoarthritic chondrocytes were inhibited(P0.01 or P0.05).All these changes showed a concentration-dependent effect.④DHEA with a concentration of 50 μmol/L improved TIMP-1 synthesis inhibited by IL-1β(P0.05).Conclusion: Dehydroepiandrosterone can reverse the degenerative changes of human chondrocytes induced by IL-1β in vitro.