摘要
目的使用IL-1β/TNF-α诱导小鼠终板软骨(CEP)细胞退变,探索褪黑素对该模式下细胞退变的治疗机制。方法使用组织培养法培养小鼠CEP细胞并进行鉴定,通过鉴定的细胞用于后续实验。用IL-1β/TNF-α刺激CEP细胞模拟炎性环境,使用免疫荧光染色和Western blotting检测细胞整合素β1和Ⅱ型胶原的表达水平,评估炎性环境下细胞与细胞外基质黏附相关分子的变化。然后,使用同样方法检测CEP细胞ki67的表达,以及通过检测细胞黏附率评估细胞的活性和黏附能力。最后,使用上述方法检测褪黑素是否可以缓解IL-1β/TNF-α刺激的CEP细胞退变。结果经甲苯胺蓝和Ⅱ型胶原染色确定培养的细胞为CEP细胞,免疫荧光染色和Western blotting结果显示IL-1β/TNF-α会导致细胞整合素β1、Ⅱ型胶原和ki67的表达水平降低(均P<0.01),细胞黏附率下降(P<0.01),但是这些病理变化可以被褪黑素抑制(P<0.05或P<0.01)。结论褪黑素可以通过增强细胞黏附缓解IL-1β/TNF-α诱导的小鼠CEP细胞退变。
Objective To use IL-1β/TNF-αto induce cartilage endplate(CEP)chondrocyte degeneration in mice and explore the therapeutic mechanism of melatonin.Methods The CEP chondrocytes of mice were isolated and cultured in the method of tissue culture,and the identified cells were used for subsequent experiments.IL-1β/TNF-αwas used to stimulate CEP chondrocytes,simulating the inflammatory environment in vivo.Immunofluorescence staining and Western blotting were used to detect the expression levels of integrinβ1 and collagenⅡ,and to evaluate the changes of molecules related to cell-extracellular matrix adhesion in inflammatory environment.Then,we used the same methods to detect the expression of ki67 in CEP chondrocytes,and detected the rate of cell adhesion to assess the cell activity and cell adhesion ability.Finally,we used methods mentioned above to detect whether melatonin could alleviate CEP chondrocytes degeneration induced by IL-1β/TNF-α.Results The cultured cells were identified as CEP chondrocytes by toluidine blue and collagenⅡstaining.Results of immunofluorescence staining and Western blotting showed that IL-1β/TNF-αcould reduce the expression levels of integrinβ1,collagenⅡand ki67,as well as the rate of cell adhesion(all P<0.01),while these pathological changes could be inhibited by melatonin(P<0.05 or P<0.01).Conclusion Melatonin can alleviate IL-1β/TNF-α-induced CEP chondrocyte degeneration in mice by enhancing cell adhesion.
作者
全惠琳
郇宇
胡学昱
QUAN Huilin;HUAN Yu;HU Xueyu(Department of Orthopedics,Xijing Hospital,Air Force Medical University,Xi'an 710032,China;Department of Neurosurgery,Xijing Hospital,Air Force Medical University,Xi'an 710032,China)
出处
《空军军医大学学报》
CAS
2022年第4期444-449,共6页
Journal of Air Force Medical University
基金
陕西省重点研发计划项目(2021ZDLSF02-10)
空军军医大学军事医学“珠峰工程”项目(2018RCFC02)
空军军医大学飞行人员作战效能提升航空医学重大问题科技攻关项目(2019ZTA12)。