目的:探讨肿瘤坏死因子相关凋亡诱导配体(Tumor necrosis factor related apoptosis-inducingligand,TRAIL)与化疗药物氟尿嘧啶(5-FU)、顺铂(DDP)联合使用诱导喉癌HEP-2细胞株凋亡。方法:不同浓度的TRAIL、5-FU、DDP单独及TRAIL与5-FU、...目的:探讨肿瘤坏死因子相关凋亡诱导配体(Tumor necrosis factor related apoptosis-inducingligand,TRAIL)与化疗药物氟尿嘧啶(5-FU)、顺铂(DDP)联合使用诱导喉癌HEP-2细胞株凋亡。方法:不同浓度的TRAIL、5-FU、DDP单独及TRAIL与5-FU、DDP联合处理HEP-2细胞,用四甲基偶氮唑蓝(MTT)法检测其细胞毒作用,荧光显微镜下观察并计算凋亡率,流式细胞仪(FCM)检测细胞凋亡率。结果:HEP-2细胞对低毒性的TRAIL不敏感,对化疗药物相对敏感,TRAIL与低毒性的化疗药物联合作用于细胞后可增强杀伤效应。结论:TRAIL与低毒性的5-Fu、DDP联用表现出高效的杀灭肿瘤细胞作用。展开更多
BACKGROUND Massive hepatocyte death is the core event in acute liver failure(ALF).Gasdermin D(GSDMD)-mediated pyroptosis is a type of highly inflammatory cell death.However,the role of hepatocyte pyroptosis and its me...BACKGROUND Massive hepatocyte death is the core event in acute liver failure(ALF).Gasdermin D(GSDMD)-mediated pyroptosis is a type of highly inflammatory cell death.However,the role of hepatocyte pyroptosis and its mechanisms of expanding inflammatory responses in ALF are unclear.AIM To investigate the role and mechanisms of GSDMD-mediated hepatocyte pyroptosis through in vitro and in vivo experiments.METHODS The expression of pyroptosis pathway-associated proteins in liver tissues from ALF patients and a hepatocyte injury model was examined by Western blot.GSDMD short hairpin RNA(shRNA)was used to investigate the effects of downregulation of GSDMD on monocyte chemotactic protein 1(MCP1)and its receptor CC chemokine receptor-2(CCR2)in vitro.For in vivo experiments,we used GSDMD knockout mice to investigate the role and mechanism of GSDMD in a D-galactose/lipopolysaccharide(D-Galn/LPS)-induced ALF mouse model.RESULTS The levels of pyroptosis pathway-associated proteins in liver tissue from ALF patients and a hepatocyte injury model increased significantly.The level of GSDMD-N protein increased most obviously(P<0.001).In vitro,downregulation of GSDMD by shRNA decreased the cell inhibition rate and the levels of MCP1/CCR2 proteins(P<0.01).In vivo,GSDMD knockout dramatically eliminated inflammatory damage in the liver and improved the survival of DGaln/LPS-induced ALF mice(P<0.001).Unlike the mechanism of immune cell pyroptosis that involves releasing interleukin(IL)-1βand IL-18,GSDMDmediated hepatocyte pyroptosis recruited macrophages via MCP1/CCR2 to aggravate hepatocyte death.However,this pathological process was inhibited after knocking down GSDMD.CONCLUSION GSDMD-mediated hepatocyte pyroptosis plays an important role in the pathogenesis of ALF,recruiting macrophages to release inflammatory mediators by upregulating MCP1/CCR2 and leading to expansion of the inflammatory responses.GSDMD knockout can reduce hepatocyte death and inflammatory responses,thus alleviating ALF.展开更多
文摘目的:探讨肿瘤坏死因子相关凋亡诱导配体(Tumor necrosis factor related apoptosis-inducingligand,TRAIL)与化疗药物氟尿嘧啶(5-FU)、顺铂(DDP)联合使用诱导喉癌HEP-2细胞株凋亡。方法:不同浓度的TRAIL、5-FU、DDP单独及TRAIL与5-FU、DDP联合处理HEP-2细胞,用四甲基偶氮唑蓝(MTT)法检测其细胞毒作用,荧光显微镜下观察并计算凋亡率,流式细胞仪(FCM)检测细胞凋亡率。结果:HEP-2细胞对低毒性的TRAIL不敏感,对化疗药物相对敏感,TRAIL与低毒性的化疗药物联合作用于细胞后可增强杀伤效应。结论:TRAIL与低毒性的5-Fu、DDP联用表现出高效的杀灭肿瘤细胞作用。
基金Supported by the National Natural Science Foundation of China,No.81570543 and No.81560104
文摘BACKGROUND Massive hepatocyte death is the core event in acute liver failure(ALF).Gasdermin D(GSDMD)-mediated pyroptosis is a type of highly inflammatory cell death.However,the role of hepatocyte pyroptosis and its mechanisms of expanding inflammatory responses in ALF are unclear.AIM To investigate the role and mechanisms of GSDMD-mediated hepatocyte pyroptosis through in vitro and in vivo experiments.METHODS The expression of pyroptosis pathway-associated proteins in liver tissues from ALF patients and a hepatocyte injury model was examined by Western blot.GSDMD short hairpin RNA(shRNA)was used to investigate the effects of downregulation of GSDMD on monocyte chemotactic protein 1(MCP1)and its receptor CC chemokine receptor-2(CCR2)in vitro.For in vivo experiments,we used GSDMD knockout mice to investigate the role and mechanism of GSDMD in a D-galactose/lipopolysaccharide(D-Galn/LPS)-induced ALF mouse model.RESULTS The levels of pyroptosis pathway-associated proteins in liver tissue from ALF patients and a hepatocyte injury model increased significantly.The level of GSDMD-N protein increased most obviously(P<0.001).In vitro,downregulation of GSDMD by shRNA decreased the cell inhibition rate and the levels of MCP1/CCR2 proteins(P<0.01).In vivo,GSDMD knockout dramatically eliminated inflammatory damage in the liver and improved the survival of DGaln/LPS-induced ALF mice(P<0.001).Unlike the mechanism of immune cell pyroptosis that involves releasing interleukin(IL)-1βand IL-18,GSDMDmediated hepatocyte pyroptosis recruited macrophages via MCP1/CCR2 to aggravate hepatocyte death.However,this pathological process was inhibited after knocking down GSDMD.CONCLUSION GSDMD-mediated hepatocyte pyroptosis plays an important role in the pathogenesis of ALF,recruiting macrophages to release inflammatory mediators by upregulating MCP1/CCR2 and leading to expansion of the inflammatory responses.GSDMD knockout can reduce hepatocyte death and inflammatory responses,thus alleviating ALF.
