胡桃醌通过Caspase-3/GSDME信号通路诱导骨肉瘤细胞焦亡  

Juglone induces pyroptosis of osteosarcoma cells through the Caspase-3/GSDME signaling pathway

作　　者：赵杰瑞 张钰涵 王馨 郭玉苗 陈姝彤 王一 吴其标[1] 彭鹏 ZHAO Jierui;ZHANG Yuhan;WANG Xin;GUO Yumiao;CHEN Shutong;WANG Yi;WU Qibiao;PENG Peng(Faculty of Chinese Medicine,Macao University of Science and Technology,Macao 999078,China;Affiliated Hospital ofFaculty of Chinese Medicine,Macao University of Science and Technology,Zhuhai 519003,China;Affiliated Hospital ofChangchun University of Traditional Chinese Medicine,Changchun 130012,China;School of Basic Medicine of BeihuaUniversity,Jilin 132000;Medical School of Yanbian University,Yanbian 133000,China)

机构地区：[1]澳门科技大学中医药学院,中国澳门999078 [2]澳门科技大学中医药学院附属珠海医院,广东珠海519003 [3]长春市中医药大学附属医院,吉林长春130012 [4]北华大学基础医学院,吉林吉林132000 [5]延边大学医学院,吉林延边133000

出　　处：《广东药科大学学报》2024年第5期8-14,共7页Journal of Guangdong Pharmaceutical University

基　　金：广东省医学科学技术研究基金资助项目(A2021004);国家自然科学基金委员会-澳门科学技术发展基金联合科研资助项目(0048/2023/AFJ);澳门科学技术发展基金资助项目(0098/2021/A2)。

摘　　要：目的探讨胡桃醌诱发焦孔素E(Gasdermin E,GSDME)依赖性细胞焦亡对骨肉瘤细胞(U2OS细胞)增殖的影响及相关机制。方法体外培养U2OS细胞,用CCK-8法检测不同浓度胡桃醌处理24 h后对细胞存活率的影响,倒置显微镜观察胡桃醌处理后U2OS细胞形态的变化,流式细胞术检测线粒体膜电位的变化,乳酸脱氢酶(LDH)释放实验检测细胞的LDH释放量,Western blot检测细胞中凋亡相关蛋白(Bax、Bcl-2、Parp1)以及GSDME、GSDME-N、caspase-3、cleave-Caspase-3相关蛋白水平变化,酶联免疫吸附分析(ELISA)检测细胞因子白介素IL-1β和IL-18。结果与对照组相比,不同浓度胡桃醌明显抑制细胞增殖(P<0.05,P<0.01),IC50为19.4μmol/L;倒置显微镜下观察到胡桃醌处理后的细胞出现肿胀、吐泡现象;诱导U2OS细胞线粒体膜电位降低,并使LDH释放增加(P<0.05,P<0.01);ELISA实验表明胡桃醌可以上调IL-1β和IL-18的表达水平;Western blot实验结果显示,胡桃醌能明显上调Bax和Parp、cleave-Caspase-3、GSDME-N的表达,降低BCL-2的表达。结论胡桃醌可抑制骨肉瘤细胞增殖,其机理可能是通过激活Bax/Caspase-3/GSDME信号通路介导细胞焦亡发挥作用。Objective To investigate the effect of gasdermin E(GSDME)-induced pyroptosis on the proliferation of osteosarcoma cells(U2OS cells)and the related mechanism.Methods U2OS cells were cultured in vitro.The CCK-8 method was used to detect the effect of juglone treatment at different concentrations on the cell survival rate for 24 h.The morphological changes in U2OS cells after juglone treatment were observed by inverted microscopy,mitochondrial membrane potential changes were detected by flow cytometry,and LDH release was detected by LDH release assay.Western blot was used to analyze apoptosis-related proteins(Bax,Bcl-2,and Parp1)and GSDME,GSDME-N,caspase-3,and cleaved caspase-3-related proteins.The cytokines IL-1βand IL-18 were detected by enzyme-linked immunosorbent assay(ELISA).Results Compared with the control group,juglone at different concentrations significantly inhibited cell proliferation(P<0.05,P<0.01),and the IC50 was 19.4μmol/L.The swelling and exhalation of juglone-treated cells were observed under an inverted microscope.The mitochondrial membrane potential of U2OS cells was decreased,and LDH release was increased(P<0.05,P<0.01).ELISA showed that juglone upregulated the expression levels of IL-1βand IL-18.Western blot results showed that juglone significantly upregulated the expression of Bax and Parp1,cleaved caspase-3,and GSDME-N and decreased the expression of BCL-2.Conclusion Juglone can inhibit the proliferation of osteosarcoma cells,and its mechanism may be the activation of the Bax/Caspase-3/GSDME signaling pathway mediated by pyroptosis.

关 键 词：胡桃醌 骨肉瘤 U2OS细胞 Caspase-3/GSDME 细胞焦亡 

分 类 号：R285.5[医药卫生—中药学]