丹参酮ⅡA下调NF-κB信号通路发挥对IL-1β所致软骨细胞损伤的保护作用  

TanshinoneⅡ-A Plays a Protective Effect on Chondrocyte Damage Induced by IL-1βby Down-regulating the NF-κB Signaling Pathway

作　　者：洪瑛 晏为华 刘修树[2] HONG Ying;YAN Wei-hua;LIU Xiu-shu(Hanshan County People′s Hospital,Hanshan 238100,China;Hefei Vocational and Technical College,Chaohu 238000,China)

机构地区：[1]含山县人民医院,安徽含山238100 [2]合肥职业技术学院,安徽巢湖238000

出　　处：《海峡药学》2021年第6期4-8,共5页Strait Pharmaceutical Journal

基　　金：国家自然科学基金青年项目(NO:81603523)。

摘　　要：目的探究丹参酮ⅡA(TanⅡA)对IL-1β所致炎性损伤软骨细胞保护作用的分子机制。方法以酶消化法分离得到的小鼠原代软骨细胞为研究对象,通过CCK-8法检测IL-1β和TanⅡA对软骨细胞增殖活性的影响,确定IL-1β与TanⅡA的作用浓度与作用时间。选择一定浓度的IL-1β处理软骨细胞构建炎性损伤模型,用不同浓度的TanⅡA进行共干预,实验分为对照组、炎性损伤模型组(IL-1β刺激)、药物治疗组(不同浓度TanⅡA干预)。通过Real-time PCR分别检测各组NF-κB信号通路关键转录因子TRAF6、RelA mRNA表达情况,Western Blot分别检测各组NF-κB信号通路关键转录因子TRAF6、p65、p-p65蛋白表达情况,分析IL-1β对软骨细胞NF-κB信号通路的促进作用,以及TanⅡA对其的抑制作用。结果≤10 ng·mL^(-1)浓度的IL-1β对软骨细胞增殖具有一定促进作用(P<0.01),<300μmol·L^(-1)浓度的TanⅡA对炎性损伤软骨细胞增殖无显著影响,而≥300μmol·L^(-1)浓度的TanⅡA对炎性损伤软骨细胞增殖具有抑制作用(P<0.05)。10 ng·mL^(-1)IL-1β分别作用软骨细胞48 h、72 h后,能够明显激活NF-κB信号通路TRAF6、RelA mRNA(P<0.01)与TRAF6、p65、p-p65蛋白的表达(P<0.01),而当用50、100、200μmol·L^(-1)的TanⅡA与IL-1β同时处理细胞时,TanⅡA能够显著抑制IL-1β刺激对NF-κB信号通路的激活作用(P<0.01)。结论TanⅡA通过抑制NF-κB信号通路激活来缓解IL-1β对软骨细胞的炎性损伤,起到保护软骨细胞的作用。OBJECTIVE To investigate the molecular mechanism of tanshinoneⅡA(TanⅡA)in protecting chondrocytes from inflammatory injury induced by IL-1β.METHODS The primary chondrocytes were isolated by enzyme digestion.The effects of IL-1βand TanⅡA on the proliferation of chondrocytes were detected by CCK-8,and the concentration and time of action of IL-1βand TanⅡA on chondrocytes were determined.The chondrocytes were treated with certain concentration of IL-1βto construct inflammatory injury model.The experimental groups were divided into control group,inflammatory injury model group(IL-1βstimulation)and drug treatment group(TanⅡA with different concentrations).The mRNA expression of key transcription factor TRAF6,RelA in NF-κB signaling pathway was detected by Real-time PCR.And western Blot was used to detect the expression of key transcription factor TRAF6,p65,p-p65 in NF-κB signaling pathway in each group.To analyze the promoting effect of IL-1βon NF-κB signaling pathway in chondrocytes and the inhibitory effect of TanⅡA on it.RESULTS IL-1β(≤10 ng·mL^(-1))could promote the proliferation of chondrocytes(P<0.01),and TanⅡA(<300μmol·L^(-1))had no significant effect on the proliferation of inflammatory injured chondrocytes.In contrast,the high concentration of TanⅡA(≥300μmol·L^(-1))inhibited the proliferation of inflammatory injured chondrocytes(P<0.05).When chondrocytes were treated with 10 ng·mL^(-1)IL-1βfor 48 h,the mRNA expression of TRAF6,RelA in NF-κB signaling pathway was significantly activated(P<0.01).And the protein expression of TRAF6,p-p65 in NF-κB signaling pathway were also significantly increased after 72 h treatment(P<0.01).When treated with 50μmol·L^(-1),100μmol·L^(-1),200μmol·L^(-1)TanⅡA and IL-1β,respectively,TanⅡA could significantly inhibit the activation of NF-κB signaling pathway induced by IL-1β(P<0.01).CONCLUSION TanⅡA protects chondrocytes by inhibiting the activation of NF-κB signaling pathway to alleviate the inflammatory injury of chondrocytes

关 键 词：骨性关节炎 软骨细胞 炎性损伤 丹参酮ⅡA NF-ΚB 

分 类 号：R965[医药卫生—药理学]