Myocardial ischemia(MI)is a pathophysiological condition in which the myocardium is unable to maintain normal cardiac function due to insufficient coronary artery blood and oxygen supply,as well as abnormal myocardial...Myocardial ischemia(MI)is a pathophysiological condition in which the myocardium is unable to maintain normal cardiac function due to insufficient coronary artery blood and oxygen supply,as well as abnormal myocardial energy metabolism[1].Ginsenoside Rbi(Rbi),one of the most abundant natural ingredients in ginseng and Panax notoginseng,has been proven to protect the heart from MI/reperfusion injury(RI)[2].展开更多
Infections of many viruses induce caspase activation to regulate multiple cellular pathways,including programmed cell death,immune signaling and etc.Characterizations of caspase cleavage sites and substrates are impor...Infections of many viruses induce caspase activation to regulate multiple cellular pathways,including programmed cell death,immune signaling and etc.Characterizations of caspase cleavage sites and substrates are important for understanding the regulation mechanisms of caspase activation.Here,we identified and analyzed a novel caspase cleavage motif AEAD,and confirmed its caspase dependent cleavage activity in natural substrate,such as nitric oxide-associated protein 1(NOA1).Fusing the enhanced green fluorescent protein(EGFP)with the mitochondrial marker protein Tom20 through the AEAD motif peptide localized EGFP to the mitochondria.Upon the activation of caspase triggered by Sendai virus(SeV)or herpes simplex virus type 1(HSV-1)infection,EGFP diffusely localized to the cell due to the caspase-mediated cleavage,thus allowing visual detection of the virusinduced caspase activation.An AEAD peptide-derived inhibitor Z-AEAD-FMK were developed,which significantly inhibited the activities of caspases-1,-3,-6,-7,-8 and-9,exhibiting a broad caspase inhibition effect.The inhibitor further prevented caspases-mediated cleavage of downstream substrates,including BID,PARP1,LMNA,pro-IL-1β,pro-IL-18,GSDMD and GSDME,protecting cells from virus-induced apoptotic and pyroptotic cell death.Together,our findings provide a new perspective for the identification of novel caspase cleavage motifs and the development of new caspase inhibitors and anti-inflammatory drugs.展开更多
Background:Paclitaxel is a compound derived from Pacific yew bark that induces various cancer cell apoptosis.However,whether it also has anticancer activities in KOSC3 cells,an oral cancer cell line,is unclear.Methods:...Background:Paclitaxel is a compound derived from Pacific yew bark that induces various cancer cell apoptosis.However,whether it also has anticancer activities in KOSC3 cells,an oral cancer cell line,is unclear.Methods:3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide,flow cytometry,and western blotting assays were carried out to assess cell viability,subG1 phase of the cell cycle,and apoptosis-related protein expression,respectively.Results:Ourfindings indicate that paclitaxel could inhibit cell viability and increase the expression of apoptotic markers,including plasma membrane blebbing and the cleavage of poly ADP-ribose polymerase in KOSC3 cells.Also,the treatment with paclitaxel remarkably elevated the percentage of the subG1 phase in KOSC3 cells.In addition,treatment with a pan-caspase inhibitor could recover paclitaxel-inhibited cell viability.Moreover,caspase-8,caspase-9,caspase-7,and BH3 interacting domain death agonist(Bid)were activated in paclitaxel-treated KOSC3 cells.Conclusions:Paclitaxel induced apoptosis through caspase cascade in KOSC3 cells.展开更多
目的:探讨顺铂(顺式二氨二氯铂)(c i sdichlorodiamineplatinum,DDP)对人结肠癌Caco-2细胞凋亡及相关蛋白Bcl-2、Caspase9、Caspase3表达的影响.方法:体外培养结肠癌Caco-2细胞;检测不同浓度DDP干预下MTT染色的A值,判定其对人结肠癌细...目的:探讨顺铂(顺式二氨二氯铂)(c i sdichlorodiamineplatinum,DDP)对人结肠癌Caco-2细胞凋亡及相关蛋白Bcl-2、Caspase9、Caspase3表达的影响.方法:体外培养结肠癌Caco-2细胞;检测不同浓度DDP干预下MTT染色的A值,判定其对人结肠癌细胞恶性增殖的影响;不同浓度D D P对人结肠癌细胞凋亡的影响使用流式细胞仪进行检测,Western blot检测不同浓度DDP对人结肠癌细胞内Bcl-2蛋白的表达;应用分光光度法检测不同浓度DDP对人结肠癌细胞内Caspase9、Caspase3蛋白活性的影响.结果:(1)癌细胞增殖结果显示,在一定的作用时间范围内,Caco-2细胞存活率与DDP浓度呈负相关,具有剂量依赖性,其中4.000、2.000μg/m L干预的各组细胞抑制率最显著,差异均有统计学意义(P<0.05).而低剂量组0.250、0.125μg/m L及对照DMSO干预组细胞存活率比较差异均无统计学意义(P>0.05),其细胞存活率明显高于其他各浓度组,差异有统计学意义(P<0.01);(2)流式细胞仪检测结果显示:DDP能诱导Caco-2细胞的凋亡,其诱导凋亡的效果呈时间和剂量性依赖;(3)Western blot检测结果显示:Bcl-2蛋白在结肠癌Caco-2细胞中低表达,DDP干预Caco-2细胞72 h后,与对照组比较,Bcl-2蛋白表达明显下降(P<0.05).Caspase9、Caspase3在Caco-2细胞中低表达,DDP干预Caco-2细胞48、72 h后,与对照组比较,Caspase9、Caspase3表达明显升高(P<0.01).结论:顺铂可以抑制Caco-2细胞增殖、诱导Caco-2细胞凋亡,其机制可能与活化Caco-2细胞中Caspase9、Caspase3蛋白以及抑制Bcl-2蛋白表达有关.展开更多
文摘Myocardial ischemia(MI)is a pathophysiological condition in which the myocardium is unable to maintain normal cardiac function due to insufficient coronary artery blood and oxygen supply,as well as abnormal myocardial energy metabolism[1].Ginsenoside Rbi(Rbi),one of the most abundant natural ingredients in ginseng and Panax notoginseng,has been proven to protect the heart from MI/reperfusion injury(RI)[2].
基金supported by the National Key R&D Program of China(2021YFC2300700)National Science and Technology Major Project(No.2018ZX10101004001005)National Natural Science Foundation of China(numbers 32070179).We thank Dr.Qinxue Hu(Wuhan Institute of Virology)and Dr.Yuchen Xia(Wuhan University)for help with materials.We thank Ding Gao from Center for Instrumental Analysis and Metrology at Wuhan Institute of Virology for his help with the Leica confocal microscope and the Operetta.
文摘Infections of many viruses induce caspase activation to regulate multiple cellular pathways,including programmed cell death,immune signaling and etc.Characterizations of caspase cleavage sites and substrates are important for understanding the regulation mechanisms of caspase activation.Here,we identified and analyzed a novel caspase cleavage motif AEAD,and confirmed its caspase dependent cleavage activity in natural substrate,such as nitric oxide-associated protein 1(NOA1).Fusing the enhanced green fluorescent protein(EGFP)with the mitochondrial marker protein Tom20 through the AEAD motif peptide localized EGFP to the mitochondria.Upon the activation of caspase triggered by Sendai virus(SeV)or herpes simplex virus type 1(HSV-1)infection,EGFP diffusely localized to the cell due to the caspase-mediated cleavage,thus allowing visual detection of the virusinduced caspase activation.An AEAD peptide-derived inhibitor Z-AEAD-FMK were developed,which significantly inhibited the activities of caspases-1,-3,-6,-7,-8 and-9,exhibiting a broad caspase inhibition effect.The inhibitor further prevented caspases-mediated cleavage of downstream substrates,including BID,PARP1,LMNA,pro-IL-1β,pro-IL-18,GSDMD and GSDME,protecting cells from virus-induced apoptotic and pyroptotic cell death.Together,our findings provide a new perspective for the identification of novel caspase cleavage motifs and the development of new caspase inhibitors and anti-inflammatory drugs.
基金The present study was supported by the National Science and Technology Council,Taiwan(MOST-107-2320-B-471-001 to YYL and MOST-110-2320-B-006-025-MY3 to BMH)by An Nan Hospital(ANHRF111-55 to TCC and BMH).
文摘Background:Paclitaxel is a compound derived from Pacific yew bark that induces various cancer cell apoptosis.However,whether it also has anticancer activities in KOSC3 cells,an oral cancer cell line,is unclear.Methods:3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide,flow cytometry,and western blotting assays were carried out to assess cell viability,subG1 phase of the cell cycle,and apoptosis-related protein expression,respectively.Results:Ourfindings indicate that paclitaxel could inhibit cell viability and increase the expression of apoptotic markers,including plasma membrane blebbing and the cleavage of poly ADP-ribose polymerase in KOSC3 cells.Also,the treatment with paclitaxel remarkably elevated the percentage of the subG1 phase in KOSC3 cells.In addition,treatment with a pan-caspase inhibitor could recover paclitaxel-inhibited cell viability.Moreover,caspase-8,caspase-9,caspase-7,and BH3 interacting domain death agonist(Bid)were activated in paclitaxel-treated KOSC3 cells.Conclusions:Paclitaxel induced apoptosis through caspase cascade in KOSC3 cells.
