姜黄素通过调控DUSP1/p38 MAPK通路减轻IL-1β诱导的软骨细胞损伤  

Curcumin attenuates IL-1β-induced chondrocyte damage by modulating the DUSP1/p38 MAPK pathway

作　　者：宋飞 范学菲[3] 刘楠楠 陈苏环 江敏 陈广祎 陈唔奇 陈晓宇 周剑[1] Song Fei;Fan Xuefei;Liu Nannan;Chen Suhuan;Jiang Min;Chen Guangyi;Chen Wuqi;Chen Xiaoyu;Zhou Jian(Dept of Orthopaedics,The First Affiliated Hospital of Anhui Medical University,Hefei230022;Dept of Orthopaedics,No.901 Hospital,PLA,Hefei230031;Dept of Histology and Embryology,Anhui Medical University,Hefei230022;Dept of Clinical Medicine,Anhui Medical University,Hefei230022)

机构地区：[1]安徽医科大学第一附属医院骨科,合肥230022 [2]中国人民解放军联勤保障部队第901医院骨科,合肥230031 [3]安徽医科大学组织胚胎学教研室,合肥230022 [4]安徽医科大学临床医学系,合肥230022

出　　处：《安徽医科大学学报》2024年第11期1903-1910,共8页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金项目(编号:81373421);安徽医科大学“早期接触科研”项目(编号:2022-ZQKY-19、2023-ZQKY-093)。

摘　　要：目的探讨姜黄素(Cur)对IL-1β诱导的软骨损伤的抑制作用,并研究DUSP1/p38 MAPK信号通路在上述过程中的调控机制关系。方法将软骨细胞(C28/I2)及骨关节炎患者术后原代软骨细胞分为对照组、实验组,实验组应用IL-1β(10μg/L)行炎性诱导处理后,应用不同浓度的Cur(0、10、20、40、60、80μmol/L)处理,细胞活力测定(CCK-8)细胞增殖抑制率,流式细胞实验检测细胞凋亡率,实时荧光定量PCR(qRT-PCR)、蛋白质印迹法(Western blot)以及免疫荧光实验检测II型胶原α1链(CollagenⅡ)、基质金属肽酶13(MMP13)、白细胞介素-1β(IL-1β)、BCL2相关X的蛋白质(Bax)、B淋巴细胞瘤-2(Bcl-2)、双特异性磷酸酶1(DUSP1)、p38丝裂原活化蛋白激酶(p38)、磷酸化p38丝裂原活化蛋白激酶(p-p38)的RNA和蛋白的表达水平。使用DUSP1干扰RNA和p38 MAPK通路抑制剂(SB)进一步验证DUSP1/p38 MAPK轴在Cur抑制软骨细胞氧化应激、凋亡与炎症中的作用。结果Cur显著抑制IL-1β诱导的软骨细胞活力的下降,显著降低了软骨细胞的凋亡与炎症水平,Cur抑制了MMP13、IL-1β、Bax、p-p38蛋白的表达,而CollagenⅡ、Bcl-2、DUSP1蛋白的表达则明显增加;IL-1β和干扰RNA沉默DUSP1并激活了p38通路,Cur则抑制p38通路的激活;使用p38 MAPK通路抑制剂可减轻细胞炎症。结论Cur通过促进DUSP1蛋白的表达,抑制p38 MAPK通路的激活,从而达到减轻IL-1β所诱导的软骨细胞凋亡与炎症的作用。Objective To investigate the inhibitory effect of curcumin(Cur)on IL-1β-induced cartilage damage and to study the relationship between the regulatory mechanisms of the DUSP1/p38 MAPK signalling pathway in the above process.Methods Chondrocytes(C28/I2)and postoperative primary chondrocytes from osteoarthritis patients were divided into control and experimental groups,and the experimental group was treated with different concentrations of Cur(0,10,20,40,60,80μmol/L)after applying the inflammatory induction treatment with IL-1β(10μg/L).The cell proliferation inhibition rate was determined by cell viability assay(CCK-8),the apoptosis rate was detected by flow cytometry assay.Real-time fluorescence quantitative PCR(qRT-PCR),Western blot,and immunofluorescence assay were used to detect type II collagenα1 chain(CollagenⅡ),matrix metallopeptidase 13(MMP13),interleukin-1β(IL-1β),BCL2-related X protein(Bax),B lymphocytoma-2(Bcl-2),dual-specificity phosphatase 1(DUSP1),p38 mitogen-activated protein kinase(p38),and phosphorylated p38 mitogen-activated protein kinase(p-p38)RNA and protein expression levels.The role of the DUSP1/p38 MAPK axis in the inhibition of chondrocyte oxidative stress,apoptosis and inflammation by Cur was further validated using DUSP1 interfering RNA and p38 MAPK pathway inhibitor(SB).Results Cur significantly inhibited the IL-1β-induced decrease in chondrocyte viability and significantly reduced the levels of oxidative stress,apoptosis,and inflammation in chondrocytes;Cur inhibited the expression of MMP13,IL-1β,Bax,and p-p38 proteins,while the expression of Collagen II,Bcl-2,and DUSP1 proteins significantly increased;IL-1βand interfering RNA silencing DUSP1 activated the p38 pathway,while Cur inhibited the activation of the p38 pathway;the use of p38 MAPK pathway inhibitors reduced cellular inflammation.Conclusion Cur attenuates IL-1β-induced oxidative stress,apoptosis and inflammation in chondrocytes by promoting the expression of DUSP1 protein and inhibiting the activation of p38 MAPK pa

关 键 词：姜黄素 骨关节炎 软骨 DUSP1 p38 MAPK 

分 类 号：R684.3[医药卫生—骨科学]