摘要
目的:探讨β-蜕皮甾酮(β-Ecd)抑制IL-1β诱导关节软骨细胞退变的作用及机制。方法:进行关节软骨细胞的分离、培养和制备空白血清、β-Ecd含药血清。取对数生长期第三代软骨细胞按1×105接种于两块96孔酶标板中,分为4组,A组:正常软骨细胞+20%空白血清;B组:正常软骨细胞+20%β-Ecd含药血清;C组:IL-1β诱导软骨细胞+20%空白血清;D组:IL-1β诱导软骨细胞+20%β-Ecd含药血清。4组同一时间培养,并相同时间对C组和D组的正常软骨细胞加入10μg/L的IL-1β进行诱导退变,诱导时间为48 h。RT-PCR计数检测各组关节软骨细胞中胶原(I、II、X型)和软骨细胞外基质中金属蛋白酶(MMP-13、MMP-9)的表达。结果:关节软骨细胞中胶原表达结果显示:与A组比较,C组I型胶原mRNA表达升高(P<0.05),B组差异无显著性(P>0.05);与C组比较,D组差异无显著性(P>0.05)。与A组比较,B组和C组Ⅱ型胶原mRNA表达明显降低(P<0.05,P<0.01);与C组比较,D组Ⅱ型胶原mRNA表达均明显升高(P<0.01)。与A组比较,C组X型胶原mRNA表达明显升高(P<0.05),B组差异无显著性(P>0.05);与C组比较,D组差异无显著性(P>0.05)。关节软骨细胞外基质中金属蛋白酶表达结果显示:与A组比较,C组MMP-9、MMP-13 mRNA表达均明显升高(P<0.01),B组差异无显著性(P>0.05);而与C组比较,D组MMP-9、MMP-13 mRNA表达均明显降低(P<0.01)。结论:β-Ecd可能通过影响膝关节软骨细胞胶原和金属蛋白酶的表达而延缓关节软骨细胞的退变。
Objective:To investigate the role and mechanism ofβ-ecdysterone(β-ecd)on inhibition of the degeneration of articular chondrocytes induced by IL-1β.Methods:The articular chondrocytes were isolated and cultured.Blank serum andβ-ecd drug serum were prepared.The third generation of logarithmic growth chondrocytes were inoculated in two 96-well microplate plates at 1×105.All cells were divided into four groups,group A:normal chondrocytes+20%blank serum,group B:normal chondrocytes+20%β-ecd containing serum,group C:chondrocytes induced by IL-1β+20%blank serum,group D:chondrocytes induced by IL-1β+20%β-ecd containing serum.Four groups of cells were cultured at the same time,and normal chondrocytes of group C and group D were added with 10μg/L of IL-1βfor induction degeneration at the same time for 48 hours.RT-PCR was used to detect the expression of collagen(I,II,X)and matrix metalloproteinase(MMP-13,MMP-9)in articular chondrocytes.Results:Collagen expression in articular chondrocytes showed that:compared with group A,mRNA expression of collagen I was increased in group C(P<0.05),while no significant difference was found in group B(P>0.05).There was no significant difference between group D and group C(P>0.05).Compared with group A,the mRNA expression of collagenⅡwas significantly decreased in group B and group C(P<0.05,P<0.01).Compared with group C,the mRNA expression of collagenⅡwere significantly higher in group D(P<0.01).Compared with group A,mRNA expression of collagen X was significantly increased in group C(P<0.05),while no significant difference was found in group B(P>0.05).There was no significant difference between group D and group C(P>0.05).The expression of metalloproteinase in the extracellular matrix of articular chondrocytes showed that:compared with group A,the mRNA expressions of mmp-9 and mmp-13 in group C were significantly increased(P<0.01),while there was no significant difference in group B(P>0.05).Compared with group C,mRNA expressions of mmp-9 and mmp-13 in group D were significantly decreased(P<0.01).Conclusion:β-ecdysterone may delay the degeneration of articular chondrocytes by affecting the expression of collagen and metalloproteinase.
作者
汤样华
徐灿达
岳振双
熊振飞
李国松
辛大伟
曾林如
黄忠名
TANG Yanghua;XU Canda;YUE Zhenshuang;XIONG Zhenfei;LI Guosong;XIN Dawei;ZENG Linru;HUANG Zhongming(Department of Orthopedics,Xiaoshan Chinese Medicie Hospital,Hangzhou,Zhejiang 311201;Department of Orthopedics,Ganzhou People’s Hospital,Ganzhou,Jiangxi 341000)
出处
《中国中医药科技》
CAS
2020年第3期366-369,共4页
Chinese Journal of Traditional Medical Science and Technology
基金
国家自然科学基金(81702187)
杭州市科技计划引导项目(20171226Y97)
杭州市萧山区社会发展重大科技攻关政策引导项目(2017308)
杭州市萧山区社会发展重大科技计划项目(2018211)。
