CAJ | 학술논문

目的研究重组大鼠白细胞介素-1β (rrIL-β)诱导关节软骨细胞退变是否与其破坏线粒体的完整性和促进细胞内活性氧的表达有关.方法体外分离、培养大鼠软骨细胞成功后,以10 ng/ml白细胞介素(IL)-1β刺激,培养24 h后,通过Annexin V-异硫氰酸荧光素(FITC)和碘化丙啶双标及Hoechst 33342荧光染色检测细胞凋亡；以罗丹明-123荧光染色和活性氧检测试剂盒检测线粒体膜电位及细胞内活性氧浓度；用荧光素酶反应检测线粒体合成三磷酸腺苷(ATP)能力来评价线粒体功能.采用t检验进行统计分析.结果 IL-1β刺激组软骨细胞的线粒体膜电位、ATP含量及软骨细胞内活性氧浓度分别为(24±4)U,(4.1 ±0.8) pmol/106个细胞及(89±7)U,均低于对照组[分别为(86±10)U,(13.3±3.0) pmol/106个细胞及(17±3)U],差异均有统计学意义(t值分别为2.693,2.587,2.665,P值分别为0.026,0.039,0.021).IL-1β刺激能明显降低软骨细胞的线粒体膜电位及其合成ATP的能力,并且能促进细胞内活性氧的表达.结论 IL-Iβ能通过破坏软骨细胞线粒体膜的完整性和促进细胞内活性氧的表达,来诱导软骨细胞去分化,从而促进关节软骨退变.
목적 연구중조대서백세포개소-1β (rrIL-β)유도관절연골세포퇴변시부여기파배선립체적완정성화촉진세포내활성양적표체유관.방법 체외분리、배양대서연골세포성공후,이10 ng/ml백세포개소(IL)-1β자격,배양24 h후,통과Annexin V-이류청산형광소(FITC)화전화병정쌍표급Hoechst 33342형광염색검측세포조망；이라단명-123형광염색화활성양검측시제합검측선립체막전위급세포내활성양농도；용형광소매반응검측선립체합성삼린산선감(ATP)능력래평개선립체공능.채용t검험진행통계분석.결과 IL-1β자격조연골세포적선립체막전위、ATP함량급연골세포내활성양농도분별위(24±4)U,(4.1 ±0.8) pmol/106개세포급(89±7)U,균저우대조조[분별위(86±10)U,(13.3±3.0) pmol/106개세포급(17±3)U],차이균유통계학의의(t치분별위2.693,2.587,2.665,P치분별위0.026,0.039,0.021).IL-1β자격능명현강저연골세포적선립체막전위급기합성ATP적능력,병차능촉진세포내활성양적표체.결론 IL-Iβ능통과파배연골세포선립체막적완정성화촉진세포내활성양적표체,래유도연골세포거분화,종이촉진관절연골퇴변.
Objective To study the effect of interleukin (IL)-1β on mitochondria of chondrocytes and reactive oxygen species.Methods Rat chondrocytes were isolated and cultured in vitro,10 ng/ml IL-1β was added for establishing model of osteoarthritis.Then,ratio of apoptosis was surveyed by Annexin V-FITC and PI flow-cytometry.After Hoechst 33342 dyeing,morphology of nucleus was observed by fluorescence microscope.After rhodamine-123 luorescent staining applied,change of mitochondria membrane potential was observed by confocal microscopy.We evaluated the ability of ATP synthesizing in mitochondria by luciferase reaction.Content of active oxygen was observed by confocal microscopy.T test was used for statistical analysis.Results IL-1β could obviously reduce the mitochondrial membrane potential of chondrocytes and its ability to synthesize ATP,and could promote the expression of reactive oxygen species in cells.The values were changed to (24±4) U,(4.1±0.8) pmol/106 cells and (89±7) U from (86±10) U,(13.3±3.0) pmol/106 cells and (17±3) U.The difference had statistical significance.Conclusion IL-1β can induce chondrocyte differentiation by destroying the mitochondrial membrane integrity of chondrocytes and promoting the expression of reactive oxygen species.It can also promote the articular cartilage degeneration.