IL-33对RANKL诱导破骨细胞增殖和凋亡的影响  

Effects of IL-33 on the proliferation and apoptosis induced by RANKL in osteoclast cells

作　　者：冯瑶 吴素琴[3] 吴英良 FENG Yao;WU Suqin;WU Yingliang(School of Life Sciences and Biopharmaceuticals,Shenyang Pharmaceutical University,Shenyang 110016,China;Pathological Anatomy Laboratory,Liaoning Institute of Basic Medicine,Shenyang 110101,China;Immunology Laboratory,Liaoning Institute of Basic Medicine,Shenyang 110101,China)

机构地区：[1]沈阳药科大学生命科学和生物制药学院,辽宁沈阳110016 [2]辽宁省基础医学研究所病理解剖学研究室,辽宁沈阳110101 [3]辽宁省基础医学研究所免疫研究室,辽宁沈阳110101

出　　处：《沈阳药科大学学报》2021年第9期935-941,共7页Journal of Shenyang Pharmaceutical University

基　　金：辽宁省自然科学基金指导计划重点项目(20170540485)。

摘　　要：目的研究白细胞介素-33(interleukin-33,IL-33)对小鼠单核巨噬细胞RAW264.7来源的破骨细胞(osteoclast cells,OC)增殖和凋亡的影响。方法体外培养RAW264.7细胞,100μg·L^(-1)核因子κB受体活化因子配体(receptor activator of nuclear factor-κB ligand,RANKL)诱导成OC,以不同浓度IL-33与OC共同培养24 h,分为IL-33低浓度组(25μg·L^(-1))、IL-33中浓度组(50μg·L^(-1))和IL-33高浓度组(100μg·L^(-1)),同时设置Control组。抗酒石酸酸性磷酸酶(tartrate resistant acid phosphatase,TRAP)染色法观察阳性OC形成情况;CCK-8法检测细胞增殖水平;流式细胞术检测细胞凋亡及细胞周期;ELISA法检测细胞Caspase-3蛋白表达水平;Western Blot法检测细胞Bcl-2、Bax蛋白表达水平。结果与Control组相比,IL-33各浓度均能抑制OC凋亡,促进细胞增殖;IL-33各浓度均能抑制OC Caspase-3和Bax蛋白表达、促进OC Bcl-2蛋白表达。结论IL-33可能通过调节Bcl-2、Bax和Caspase-3蛋白水平促进RANKL诱导OC增殖、抑制其凋亡,这或许是类风湿关节炎骨破坏加重的可能作用机制。Objective To investigate the effects of interleukin-33(IL-33) on the proliferation and apoptosis of osteoclast cells(OC) derived from mouse mononuclear macrophages RAW264.7.Methods RAW264.7 cells were cultured in vitro and induced into OC by receptor activator of nuclear factor-κB ligand(RANKL)of 100 μg·L^(-1).Different concentrations of IL-33 were cultured with OC for 24 h, which were divided into the low-concentration group of IL-33(25 μg·L^(-1)),the medium-concentration group of IL-33(50 μg·L^(-1)),the high-concentration group of IL-33(100 μg·L^(-1)),and the control group.Tartrate resistant acid phosphatase(TRAP) staining was used to observe the formation of positive OC.Cell proliferation level was detected by CCK-8 method.Cell apoptosis and cell cycle were detected by flow cytometry.The expression level of Caspase-3 protein was detected by ELISA.The expression levels of Bcl-2 and Bax protein were detected by Western Blot.Results Compared with the control group, all the concentrations of IL-33 could inhibit the apoptosis of OC and promote the proliferation of OC;All the concentrations of IL-33 could inhibit the expression of Caspase-3 and Bax proteins of OC and promote the expression of Bcl-2 proteins of OC.Conclusion IL-33 can promote the proliferation of RANKL-induced OC and inhibit its apoptosis by regulating the levels of Bcl-2,Bax and Caspase-3 proteins, which may be a possible mechanism for the aggravation of osteodestruction in rheumatoid arthritis.

关 键 词：破骨细胞 白细胞介素-33 细胞凋亡 细胞增殖 

分 类 号：R96[医药卫生—药理学]