目的:探讨聚(ADP-核糖)聚合酶家族成员3[Poly(ADP-Ribose)Polymerase Family Member 3,PARP3]在脂多糖(Lipopolysaccharide,LPS)诱导的小胶质细胞BV2炎症及凋亡中的作用机制,为视神经损伤的临床治疗提供潜在靶点。方法:通过LPS刺激BV2...目的:探讨聚(ADP-核糖)聚合酶家族成员3[Poly(ADP-Ribose)Polymerase Family Member 3,PARP3]在脂多糖(Lipopolysaccharide,LPS)诱导的小胶质细胞BV2炎症及凋亡中的作用机制,为视神经损伤的临床治疗提供潜在靶点。方法:通过LPS刺激BV2小胶质细胞,构建体外炎症损伤模型,随后利用实时荧光定量聚合酶链反应技术、蛋白质印迹及流式细胞术等技术手段,检测PARP3的蛋白及mRNA表达水平、炎症因子的分泌以及细胞凋亡的情况。结果:LPS处理后,BV2细胞中PARP3表达水平显著上调,炎症因子分泌增加,细胞活性氧增多,细胞凋亡率增加,而抑制PARP3能显著减少这些效应。结论:PARP3在LPS诱导的BV2细胞炎症及凋亡中起关键作用,为视神经损伤治疗提供了新的潜在靶点。展开更多
Objective:Ovarian cancer(OC)ranks among the leading causes of mortality among the female cancers worldwide.Numerous studies have explored the development and progression of OC at multiple genetic regulatory levels.How...Objective:Ovarian cancer(OC)ranks among the leading causes of mortality among the female cancers worldwide.Numerous studies have explored the development and progression of OC at multiple genetic regulatory levels.However,relatively few studies have explored the impact of post-translational modifications(PTM)on OC progression,which is essential for uncovering new therapeutic targets.This study aimed to systematically identify the key PTM types involved in OCprogression,and to explore and evaluate their translational potential as therapeutic targets.Methods:First,we utilized multiple general PTM antibodies to compare gross PTM levels between normal ovarian and OC tissues from clinical females.After identifying lactylation as the PTM with the most significant differences,we selected representative samples for label-free mass spectrometry to identify specific lactylation sites.Next,we transfected A2780(OC)cells with either wild-type(WT)or mutant(K192A[Q])poly(ADP-ribose)polymerase 1(PARP1)conjugated to enhanced green fluorescent protein(EGFP)with a StrepⅡpeptide tag and assessed various cellular indexes related to cell proliferation(clonogenicity assay),migration(scratch wound healing assay),and reactive oxygen species levels.Results:Pan-lactylation was significantly upregulated in clinical OC samples,with PARP1 lactylation at K192 being one of the most common modifications.The growth and migration of A2780 cells were markedly suppressed by overexpressing PARP1-WT but not mutant PARP1.Overexpressing PARP1 significantly downregulated the phosphorylation of extracellular signal-regulated kinases 1/2(ERK1/2).Conclusion:This study uncovered a novel PTM of PARP1 in OC,lactylation,and demonstrated that lactylation at K192 is crucial in regulating OC cell growth and migration via the ERK1/2 pathway.Further investigations are required to elucidate the broader functional implications of PARP1 lactylation and its therapeutic potential.展开更多
文摘目的:探讨聚(ADP-核糖)聚合酶家族成员3[Poly(ADP-Ribose)Polymerase Family Member 3,PARP3]在脂多糖(Lipopolysaccharide,LPS)诱导的小胶质细胞BV2炎症及凋亡中的作用机制,为视神经损伤的临床治疗提供潜在靶点。方法:通过LPS刺激BV2小胶质细胞,构建体外炎症损伤模型,随后利用实时荧光定量聚合酶链反应技术、蛋白质印迹及流式细胞术等技术手段,检测PARP3的蛋白及mRNA表达水平、炎症因子的分泌以及细胞凋亡的情况。结果:LPS处理后,BV2细胞中PARP3表达水平显著上调,炎症因子分泌增加,细胞活性氧增多,细胞凋亡率增加,而抑制PARP3能显著减少这些效应。结论:PARP3在LPS诱导的BV2细胞炎症及凋亡中起关键作用,为视神经损伤治疗提供了新的潜在靶点。
文摘Objective:Ovarian cancer(OC)ranks among the leading causes of mortality among the female cancers worldwide.Numerous studies have explored the development and progression of OC at multiple genetic regulatory levels.However,relatively few studies have explored the impact of post-translational modifications(PTM)on OC progression,which is essential for uncovering new therapeutic targets.This study aimed to systematically identify the key PTM types involved in OCprogression,and to explore and evaluate their translational potential as therapeutic targets.Methods:First,we utilized multiple general PTM antibodies to compare gross PTM levels between normal ovarian and OC tissues from clinical females.After identifying lactylation as the PTM with the most significant differences,we selected representative samples for label-free mass spectrometry to identify specific lactylation sites.Next,we transfected A2780(OC)cells with either wild-type(WT)or mutant(K192A[Q])poly(ADP-ribose)polymerase 1(PARP1)conjugated to enhanced green fluorescent protein(EGFP)with a StrepⅡpeptide tag and assessed various cellular indexes related to cell proliferation(clonogenicity assay),migration(scratch wound healing assay),and reactive oxygen species levels.Results:Pan-lactylation was significantly upregulated in clinical OC samples,with PARP1 lactylation at K192 being one of the most common modifications.The growth and migration of A2780 cells were markedly suppressed by overexpressing PARP1-WT but not mutant PARP1.Overexpressing PARP1 significantly downregulated the phosphorylation of extracellular signal-regulated kinases 1/2(ERK1/2).Conclusion:This study uncovered a novel PTM of PARP1 in OC,lactylation,and demonstrated that lactylation at K192 is crucial in regulating OC cell growth and migration via the ERK1/2 pathway.Further investigations are required to elucidate the broader functional implications of PARP1 lactylation and its therapeutic potential.
