应用原代软骨细胞与SW1353软骨肉瘤细胞系建立体外骨关节炎模型的效果评价  

Evaluation of in vitro osteoarthritis models established by primary chondrocytes and SW1353 chondrosarcoma cell lines

作　　者：刘晓彤 黄悦 刘旭丹[1] 徐小磊[1] 姜梦琪[1] 杨迎春 何健宜 顾海伦[2] 刘莉[1] Liu Xiaotong;Huang Yue;Liu Xudan;Xu Xiaolei;Jiang Mengqi;Yang Yingchun;He Jianyi;Gu Hailun;Liu Li(Department of Nutrition and Food Hygiene,School of Public Health,China Medical University,Shenyang 110100;Department of Orthopedics,Shengjing Hospital,China Medical University,Shenyang 110004,China)

机构地区：[1]中国医科大学公共卫生学院营养与食品卫生教研室,沈阳110100 [2]中国医科大学附属盛京医院骨科,沈阳110004

出　　处：《中国组织化学与细胞化学杂志》2018年第5期459-464,共6页Chinese Journal of Histochemistry and Cytochemistry

基　　金：国家自然科学基金资助(81372971);沈阳市科技局项目(RC170476)

摘　　要：目的通过比较白细胞介素-1β(interleukin-1β,IL-1β)处理对原代软骨细胞与SW1353软骨肉瘤细胞系增殖活力、炎症因子与炎症通路表达水平变化的影响,为骨关节炎体外研究用细胞提供多重选择。方法免疫细胞化学法与甲苯胺蓝染色分别检测细胞中Ⅱ型胶原与蛋白多糖,鉴定所培养的原代细胞是否为软骨细胞。CCK-8法检测IL-1β(10ng/ml)处理24h、48h、72h对原代软骨细胞增殖活力的影响,IL-1β(1、10、20、40ng/ml)处理24h对SW1353软骨肉瘤细胞系增殖活力的影响。IL-1β(10ng/ml)分别处理原代软骨细胞与SW1353软骨肉瘤细胞系细胞24h后,ELISA法检测细胞培养上清中白细胞介素6(interleukin-6,IL-6)与基质金属蛋白酶-13(matrix metalloproteinase-13,MMP-13)的表达水平。Real-time PCR法检测核因子-κB(nuclear factor-κB,NF-κB)mRNA表达水平。结果所培养的原代细胞为原代软骨细胞。IL-1β(10ng/ml)处理可显著抑制原代软骨细胞增殖活力,但IL-1β(1、10、20、40 ng/ml)处理对SW1353软骨肉瘤细胞系增殖活力无明显影响。IL-1β(10ng/ml)处理可使IL-6、MMP-13表达水平及NF-κB mRNA的表达量均显著增加。结论IL-1β作用下原代软骨细胞与SW1353软骨肉瘤细胞系均可表现出骨关节炎样炎症反应,二者均可用于骨关节炎的体外实验研究。Objective To provide multiple choices for cells for in vitro studies of osteoarthritis by comparing the effect of Interleukin-1β(IL-1β)treatment on proliferation activity,inflammatory factors and the expression of inflammatory pathways in primary chondrocytes and SW1353 chondrosarcoma cell lines.Methods Immunocytochemical staining and toluidine blue staining were used to detect type II collagen and proteoglycan,respectively,and to identify whether the cultured primary cells were chondrocytes.cck-8 assay was used to detect the effects of IL-1β(10ng/ml)treatment on the proliferation of primary chondrocytes after 24h,48h and 72h and the effect of IL-1β(1,10,20,40ng/mL)treatment on the proliferation of SW1353 cell lines after 24h.After IL-1β(10ng/ml)treatment on primary chondrocytes and SW1353 cell lines for 24h,the expression of Interleukin-6(IL-6)and matrix metalloproteinase-13(MMP-13)in the cell culture medium supernatant was detected by ELISA.The expression of nuclear factor-kappa B(NF-κB)mRNA was examined by Real-time PCR.Results The results showed that the cultured cells were primary chondrocytes.IL-1β(10ng/ml)treatment could significantly inhibit the proliferation activity of primary chondrocytes,but IL-1β(1,10,20,40ng/ml)had no significant effect on the proliferation activity of SW1353 cell lines.Treatment with IL-1β(10ng/ml)significantly increased the expression level of IL-6,MMP-13 and NF-κB mRNA.Conclusion Both primary chondrocytes and SW1353 chondrosarcoma cell lines can exhibit ILosteoarthritis-like inflammatory reactions,when treated with IL-1β,and therefore can be used for in vitro experimental studies of osteoarthritis.〔Keywords

关 键 词：骨关节炎 原代软骨细胞 SW1353软骨肉瘤细胞系 核因子-ΚB 

分 类 号：R364.5[医药卫生—病理学]