蠲痹汤调控软骨细胞铁死亡治疗骨关节炎  

Juanbi decoction regulates iron death of chondrocytes to treat osteoarthritis

作　　者：高浩阳 贾庆运 吴立生[2] 王乾 孙铭远 刘玉兴 高万里[2] Gao Haoyang;Jia Qingyun;Wu Lisheng;Wang Qian;Sun Mingyuan;Liu Yuxing;Gao Wanli(School of Clinical Medicine,Shandong Second Medical University,Weifang 261053,China;Linyi People’s Hospital,Shandong Second Medical University,Linyi 276000,China)

机构地区：[1]山东第二医科大学临床医学院,潍坊261053 [2]山东第二医科大学附属临沂市人民医院,临沂276000

出　　处：《中华实验外科杂志》2024年第8期1784-1788,共5页Chinese Journal of Experimental Surgery

基　　金：山东省自然科学基金青年项目(ZR2021QH230)。

摘　　要：目的通过体外细胞模型, 探讨蠲痹汤对骨关节炎(OA)的作用机制。方法将ATDC5细胞分为4组:正常组、OA组、蠲痹汤组和Fer-1组。OA组加入10 ng/ml白细胞介素(IL)-1β诱导炎症杜尔伯科改良伊格尔(DMEM)培养基;蠲痹汤组加入IL-1β+蠲痹汤诱导炎症DMEM培养基;Fer-1组加入IL-1β+Fer-1诱导炎症DMEM培养基。进行蛋白质印迹法(Western blot)试验检测软骨细胞的肿瘤坏死因子-α(TNF-α)、诱导型一氧化氮合酶(iNOS)、环氧化酶-2(COX-2), Collagen Ⅱ、Aggrecan、基质金属蛋白酶13(MMP-13)、p53、ACSL4、SLC7A11和GPX-4的表达, 使用细胞免疫荧光染色检测GPX-4的表达。利用化学发光法检测软骨细胞的代谢产物丙二醛(MDA)、谷胱甘肽过氧化物酶(GSH)、Fe2+、线粒体内铁的含量。多组比较采用单因素方差分析。结果 Fer-1组和蠲痹汤组的TNF-α(3.282±0.261、2.255±0.253)、iNOS(5.046±0.478、3.443±0.039)、COX-2(3.484±0.333、2.450±0.467)蛋白相对表达水平低于IL-1β组(4.384±0.601、6.863±0.637、4.812±0.496), 差异有统计学意义(t=3.848、7.432、5.579、10.500、4.289、7.629, P<0.05)。Fer-1组和蠲痹汤组的MMP-13(2.738±0.224、4.276±0.663)、p53(3.364±0.898、4.289±0.936)、ACSL4(2.897±0.767、2.776±0.594)蛋白相对表达水平低于IL-1β组(5.779±0.018、7.413±1.635、6.077±0.847), 差异有统计学意义(t=10.640、5.258、4.752、3.666、6.048、6.279, P<0.05)。Fer-1组和蠲痹汤组的Collagen Ⅱ(0.752±0.143、0.609±0.073)、Aggrecan(0.663±0.139、0.570±0.022)、SLC7A11(0.623±0.037、0.400±0.034)、GPX-4(0.686±0.041、0.537±0.065)蛋白相对表达水平高于IL-1β组(0.298±0.049、0.268±0.154、0.097±0.021、0.163±0.063), 差异有统计学意义(t=4.661、3.895、4.631、3.538、23.700、13.660、12.890、9.226, P<0.05)。Fer-1组和蠲痹汤组的GPX-4(15.476±2.519、12.133±0.703)荧光强度高于IL-1β组(5.141±0.153), 差异有统计学意义(t=8.117、5.491, P<0.05)。Fer-1组和蠲痹汤组的�ObjectiveThe mechanism of action of osteoarthritis(OA)induced by Juanbi decoction is investigated in this test using an in vitro cell model.MethodsThe ATDC5 cells were divided into four groups:the normal group,theinterleukin(IL)-1βgroup,the IL-1β+Juanbi decoction group,and the IL-1β+Fer-1 group.In the IL-1βgroup,Dulbecco’s modified Eagle’s medium(DMEM)was used to induce inflammation with 10 ng/ml IL-1β.The IL-1β+Juanbi decoction group was treated with DMEM medium to induce inflammation.The IL-1β+Fer-1 set was added to induce inflammation in the DMEM medium.The expression of tumor necrosis factor-α(TNF-α),inducible nitric oxide synthase(iNOS),cyclooxygenase-2(COX-2),CollagenⅡ,Aggrecan,matrix metalloproteinase-13(MMP-13),p53,ACSL4,SLC7A11 and GPX-4 in chondrocytes was assessed via Western blotting.GPX-4 expression was evaluated through immunofluorescence staining.Chemiluminescence was utilized to measure the levels of malondialdehyde(MDA),glutathione peroxidase(GSH),Fe 2+,and mitochondrial iron in chondrocytes.One-way analysis of variance was employed to compare multiple groups.ResultsThe relative protein expression levels of TNF-α(3.282±0.261,2.255±0.253),iNOS(5.046±0.478,3.443±0.039),and COX-2(3.484±0.333,2.450±0.467)in the IL-1β+Fer-1 group and IL-1β+Juanbi decoction group were significantly lower compared to the IL-1βgroup(4.384±0.601,6.863±0.637,4.812±0.496).The differences were statistically significant with t-values of 3.848,7.432,5.579,10.5,4.289,and 7.629 respectively(P<0.05).The relative protein expression levels of MMP-13(2.738±0.224,4.276±0.663),p53(3.364±0.898,4.289±0.936),and ACSL4(2.897±0.767,2.776±0.594)in the IL-1β+Fer-1 group and IL-1β+Juanbi decoction group were significantly lower compared to the IL-1βgroup(5.779±0.018,7.413±1.635,6.077±0.847).The differences were statistically significant with t-values of 10.64,5.258,4.752,3.666,6.048,and 6.279 respectively(P<0.05).The relative protein expression levels of CollagenⅡ(0.752±0.143,0.609±0.073),Aggrecan(0

关 键 词：蠲痹汤 铁死亡 骨关节炎 

分 类 号：R285[医药卫生—中药学]