摘要
目的:探讨不同浓度地塞米松对白介素-1β(IL-1β)诱导的SW1353细胞骨关节炎细胞基质金属蛋白酶(matrix metalloproteinase,MMPs)表达水平的抑制作用。方法:取人软骨肉瘤细胞(SW1353细胞)分别置于含浓度为10^(1)~10^(8)nmol/L地塞米松培养液中,培养12 h后采用MTS法筛选适宜的地塞米松浓度范围进行后续实验。将SW1353细胞使用10 ng/mL IL-1β进行致炎造模后,给予10^(1)~10^(7)nmol/L地塞米松干预,使用实时定量-聚合酶链反应(real-time quantitative polymerase chain reaction,RT-PCR)方法检测MMP-1、MMP-3、MMP-13的mRNA表达水平。选取10^(4)nmol/L地塞米松干预后提取MMP-1、MMP-3、MMP-13蛋白,使用Western Blot法检测其蛋白表达水平。结果:MTS增殖实验结果显示,10^(1)~10^(7)nmol/L地塞米松对SW1353细胞活性无明显影响,10^(8)nmol/L地塞米松对SW1353有明显抑制作用(P<0.05),加入10 ng/mL IL-1β后对细胞增殖无明显影响。RT-PCR结果提示,IL-1β组和空白对照组比较,MMP-1、MMP-3、MMP-13 mRNA的表达水平明显升高,差异有统计学意义(P<0.05)。和IL-1β组相比,10^(2)~10^(7)nmol/L浓度地塞米松干预后,MMP-1 mRNA表达水平明显降低(P<0.05);10^(4)~10^(7)nmol/L地塞米松干预后,MMP-3 mRNA表达水平明显降低(P<0.05);10^(1)~10^(7)nmol/L地塞米松干预后,MMP-13 mRNA水平明显减少(P<0.05)。Western blot结果显示,相对于空白对照组,IL-1β组MMP-1、MMP-3、MMP-13蛋白表达水平明显增加;继续加入10^(4)nmol/L地塞米松干预后MMP-1、MMP-3、MMP-13蛋白表达水平明显降低。结论:地塞米松能够在mRNA及蛋白质水平抑制IL-1β诱导的骨关节炎细胞模型MMP-1、MMP-3、MMP-13的表达。
Objective:To investigate the inhibition effects of dexamethasone at different concentrations on the expression of MMP-1,MMP-3,and MMP-13 in SW1353 osteoarthritis cells induced by IL-1β.Methods:SW1353 cells were placed in a culture medium containing 10^(1)-10^(8)nmol/L dexamethasone,and after 12 h of culture,the appropriate intervention concentration range of dexamethasone was screened by the MTS for further experiments.After SW1353 cell models were established by adding 10 ng/mL IL-1β,10^(1)-10^(7)nmol/L dexamethasone were given for intervention,and RT-PCR was used to detect the mRNA expression levels of MMP-1,MMP-3,and MMP-13.In another experiment,MMP-1,MMP-3,and MMP-13 proteins were extracted after the intervention of 10^(4)nmol/L dexamethasone,and the expression levels of these proteins were detected with Western blot method.Results:MTS proliferation experiment results showed that 10^(1)-10^(7)nmol/L dexamethasone had no significant effects on SW1353 cell viability,while 10^(8)nmol/L dexamethasone had a significant inhibitory effect on SW1353(P<0.05).SW1353 cell viability was not obviously inhibited by adding 10 ng/mL IL-1βneither.RT-PCR results indicated that the IL-1βsignificantly increased the mRNA expression levels of MMP-1,MMP-3,and MMP-13 compared with the blank control group(P<0.05).While after intervention with 10^(2)-10^(7)nmol/L dexamethasone,the increased MMP-1 mRNA expression levels were decreased(P<0.05);after intervention with 10^(3)-10^(7)nmol/L dexamethasone,MMP-3 mRNA expression levels decreased(P<0.05);after intervention with 10-10^(7)nmol/L dexamethasone,the mRNA expression of MMP-13 decreased(P<0.05).The Western blot results showed that compared with the blank control group,the expression levels of MMP-1,MMP-3,and MMP-13 proteins in the IL-1βgroup were significantly increased;but the increased expression levels of MMP-1,MMP-3,and MMP-13 proteins were significantly reduced after the intervention of 10^(4)nmol/L dexamethasone.Conclusions:Dexamethasone can inhibit the expression of MMP-1,MMP-3,and MMP-13 in the IL-1β-induced osteoarthritis cell models at the mRNA and protein levels.
作者
陈光耀
陈嘉琪
姚传辉
徐愿
罗静
鄢泽然
陶庆文
CHEN Guang-yao;CHEN Jia-qi;YAO Chuan-hui;XU Yuan;LUO Jing;YAN Ze-ran;TAO Qing-wen(Beijing University of Traditional Chinese Medicine,Beijing 100029,China;Rheumatology Department of Traditional Chinese Medicine,China‑Japan Friendship Hospital,Beijing Key Laboratory of Immune Inflammatory Diseases,Beijing 10029,China)
出处
《海南医学院学报》
CAS
2021年第10期757-762,767,共7页
Journal of Hainan Medical University
基金
国家自然科学基金资助项目(81673941,81704050,81804042)。
