毛兰素调节CCL2⁃CCR2轴对关节软骨细胞炎性损伤的影响  

The effect of Erianin on inflammatory injury of articular chondrocytes by regulating the CCL2⁃CCR2 axis

作　　者：樊增荣 蒋勇 刘德洪 FAN Zengrong;JIANG Yong;LIU Dehong(Department of Orthopedics,Jianyang Air Separation Hospital,Jianyang 641400,China;Department of Proctology,Jianyang Air Separation Hospital,Jianyang 641400,China)

机构地区：[1]简阳空分医院骨科,四川简阳641400 [2]简阳空分医院肛肠科,四川简阳641400

出　　处：《中国骨质疏松杂志》2024年第3期335-340,354,共7页Chinese Journal of Osteoporosis

基　　金：四川省中医药信息学会科研项目(20200119)。

摘　　要：目的探讨毛兰素调节CC趋化因子配体2(CCL2)/CC趋化因子受体2(CCR2)轴对关节软骨细胞炎性损伤的影响。方法将人关节软骨细胞分为control组(正常培养)、LPS组、毛兰素低(10 nmol/L)、中(25 nmol/L)、高(50 nmol/L)剂量组、pcDNA组(转染pcDNA3.1)、pcDNA⁃CCL2组(转染pcDNA3.1⁃CCL2),实时荧光定量PCR(qRT⁃PCR)检测细胞中CCL2、CCR2 mRNA表达水平;MTT法、平板克隆实验与流式细胞仪分别检测细胞增殖和凋亡情况;qRT⁃PCR和ELISA检测IL⁃6、IL⁃1β、TNF⁃α水平;Western blot法检测细胞中增殖细胞核抗原(PCNA)、细胞抗凋亡因子B细胞淋巴瘤蛋白2(Bcl⁃2)、Bcl⁃2相关X蛋白(Bax)及CCL2/CCR2通路蛋白表达。结果与control组比较,LPS组细胞凋亡率、IL⁃6、IL⁃1β、TNF⁃αmRNA表达水平、CCL2、CCR2 mRNA及Bax、CCL2、CCR2蛋白表达水平显著升高,OD490值、集落形成数、PCNA、Bcl⁃2蛋白表达水平降低(P<0.05);与LPS组比较,毛兰素低、中、高剂量组细胞凋亡率、IL⁃6、IL⁃1β、TNF⁃αmRNA表达水平、CCL2、CCR2 mRNA及Bax、CCL2、CCR2蛋白表达水平逐渐降低,OD490值、集落形成数、PCNA、Bcl⁃2蛋白表达水平逐渐升高(P<0.05);过表达CCL2减弱了毛兰素对LPS诱导的关节软骨细胞的影响(P<0.05)。结论毛兰素可抑制LPS诱导的关节软骨细胞凋亡和炎症反应,其机制可能与抑制CCL2/CCR2通路有关。Objective To investigate the effect of Erianin on inflammatory injury of articular chondrocytes by regulating the C⁃C motif chemokine ligand 2(CCL2)/C⁃C motif chemokine receptor 2(CCR2)axis.Methods Human articular chondrocytes were grouped into control group(normal culture),LPS group,low(10nmol/L),medium(25nmol/L),and high(50nmol/L)dose groups,pcDNA group(transfected with pcDNA3.1),and pcDNA⁃CCL2 group(transfected with pcDNA3.1 CCL2).Real⁃time fluorescence quantitative PCR(qRT⁃PCR)was applied to detect the expression levels of CCL2 and CCR2 mRNA in cells.MTT assay,plate cloning assay,and flow cytometry were applied to detect cell proliferation and apoptosis,respectively.qRT⁃PCR and ELISA were applied to detect levels of IL⁃6,IL⁃1β,and TNF⁃α.Western blotting was applied to detect the expressions of proliferating cell nuclear antigen(PCNA),anti apoptotic factor B cell lymphoma protein 2(Bcl⁃2),Bcl⁃2 associated X protein(Bax),and CCL2/CCR2 pathway protein in cells.Results Compared to those in the control group,the cell apoptosis rate,IL⁃6,IL⁃1β,TNF⁃αmRNA expression levels and contents,CCL2,CCR2 mRNA,and Bax,CCL2,and CCR2 protein expression levels obvious increased in the LPS group,and the OD490 value,colony formation number,and PCNA and Bcl⁃2 protein expression levels decreased(P<0.05).Compared to those in the LPS group,the cell apoptosis rate,IL⁃6,IL⁃1β,TNF⁃αmRNA expression levels and contents,CCL2,CCR2 mRNA,and Bax,CCL2,and CCR2 protein expression levels obvious decreased in the the low,medium,and high dose Erianin groups,and the OD490 value,colony formation number,and PCNA and Bcl⁃2 protein expression levels increased(P<0.05).Overexpression of CCL2 weakened the effect of Erianin on LPS⁃induced articular chondrocytes(P<0.05).Conclusion Erianin inhibits LPS⁃induced apoptosis and inflammatory response of articular chondrocytes.The mechanism may be related to the inhibition of CCL2/CCR2 pathway.

关 键 词：毛兰素 CC趋化因子配体2 CC趋化因子受体2 关节软骨细胞 炎性损伤 

分 类 号：R684.3[医药卫生—骨科学]