PARP-1与HIV的关系

多聚（ADP-核糖）聚合酶-1[poly（ADP—ribose）polymerase-1,PARP-1]是真核细胞中存在的一种蛋白质翻译后修饰酶。而艾滋病已经蔓延到全世界各国,被传染上病毒的人数也正在大幅度上升。有研究指出,PARP-1主要在基因整合和转录两个水平调节HIV的感染。研究PARP-1与HIV的关系,可能会打开治疗艾滋病的新方向。     

Trp-tRNA synthetase bridges DNA-PKcs to PARP-1 to link IFN-γ and p53 signaling

Interferon-γ (IFN-γ) engenders strong antiproliferative responses, in part through activation of p53. However, the long-known IFN-γ-dependent upregulation of human Trp-tRNA synthetase (TrpRS), a cytoplasmic enzyme that activates tryptophan to form Trp-AMP in the first step of protein synthesis, is unexplained. Here we report a nuclear complex of TrpRS with the catalytic subunit of DNA-dependent protein kinase (DNA-PKcs) and with poly(ADP-ribose) polymerase 1 (PARP-1), the major PARP in human cells. The IFN-γ-dependent poly(ADP-ribosyl)ation of DNA-PKcs (which activates its kinase function) and concomitant activation of the tumor suppressor p53 were specifically prevented by Trp-SA, an analog of Trp-AMP that disrupted the TrpRS-DNA-PKcs-PARP-1 complex. The connection of TrpRS to p53 signaling in vivo was confirmed in a vertebrate system. These and further results suggest an unexpected evolutionary expansion of the protein synthesis apparatus to a nuclear role that links major signaling pathways.     

PARP-1与中枢神经系统疾病的关系

聚腺苷酸二磷酸核糖基聚合酶1[poly（ADP-ribose）pdymerase-1,PARP-1]是-种广泛存在于真核生物细胞核中的蛋白酶,在中枢神经系统的疾病发生中扮演着重要的角色。在外界损伤刺激下,PARP-1易被受损的DNA激活,进而通过不同途径影响神经细胞的生理功能,引起细胞炎性反应,甚至导致细胞死亡,触发中枢神经系统疾病的发生。抑制PARP-1在治疗慢性和急性的中枢神经系统疾病的作用也越来越受到重视。     

抗肿瘤靶点 PARP-1 及其抑制剂的研究进展

聚二磷酸腺苷核糖聚合酶 -1（PARP-1）参与 DNA 损伤修复,是近年来肿瘤治疗领域的热门靶点。从 PARP-1 的结构和作用机制出发, 综述 PARP-1 抑制剂的主要结构类型及优化思路,展望其应用前景和亟需解决的问题。     

PARP-1／AIF介导的细胞凋亡

PARP-1／AIF通路介导的非caspase依赖性细胞凋亡多见于缺血再灌注或某些药物引起的神经细胞死亡。PARP-1定位于细胞核,参与细胞内多种生理活动。广泛的DNA损伤引起PARP-1过度激活,进而使线粒体蛋白AIF转位至细胞核,作为DNA内切酶引发染色质凝集、DNA片段化和细胞死亡。最近,对该通路信号转导和调控机制的研究取得了快速的进展。     

Cytokine-mediated β-cell damage in PARP-1-deficient islets

Poly(ADP)-ribose polymerase (PARP) is an abundant nuclear protein that is activated by DNA damage; once active, it modifies nuclear proteins through attachment of poly(ADP)-ribose units derived from β-nicotinamide adenine dinucleotide (NAD(+)). In mice, the deletion of PARP-1 attenuates tissue injury in a number of animal models of human disease, including streptozotocin-induced diabetes. Also, inflammatory cell signaling and inflammatory gene expression are attenuated in macrophages isolated from endotoxin-treated PARP-1-deficient mice. In this study, the effects of PARP-1 deletion on cytokine-mediated β-cell damage and macrophage activation were evaluated. There are no defects in inflammatory mediator signaling or inflammatory gene expression in macrophages and islets isolated from PARP-1-deficient mice. While PARP-1 deficiency protects islets against cytokine-induced islet cell death as measured by biochemical assays of membrane polarization, the genetic absence of PARP-1 does not effect cytokine-induced inhibition of insulin secretion or cytokine-induced DNA damage in islets. While PARP-1 deficiency appears to provide protection from cell death, it fails to provide protection against the inhibitory actions of cytokines on insulin secretion or the damaging actions on islet DNA integrity.     

Association of PARP-1, NF-κB,NF-κBIA and IL-6, IL-1β and TNF-α with Graves Disease and Graves Ophthalmopathy

Background

Graves Disease (GD) is an autoimmune disorder affected by an interaction of multiple genes such as Nuclear Factor-κB (NF-κB), Nuclear Factor-κB Inhibitor (NF-κBIA), Poly (ADP-ribose) polymerase-1 (PARP-1) and cytokines like Interleukin-1β (IL-1β), Interleukin-6 (IL-6) and Tumor Necrosis Factor-α (TNF-α) and mostly accompanied by an ocular disorder, Graves Ophthalmopathy (GO). We hypothesize that there is a relationship between GD, GO, polymorphisms of inflammatory related genes and their association with cytokines, which may play important roles in autoimmune and inflammatory processes.

Subjects and methods

To confirm our hypothesis, we studied the polymorphisms and cytokine levels of 120 patients with GD and GO using PCR-RFLP and ELISA methods, respectively.

Results

We found that patients with GG genotype and carriers of G allele of PARP-1 G1672A polymorphism are at risk in the group having GD (p = 0.0007) while having GA genotype may be protective against the disease. PARP-1 C410T polymorphism was found to be associated with GO by increasing the risk by 1.7 times (p = 0.004). Another risk factor for development of GO was the polymorphism of del/ins of NFkB1 gene (p = 0.032) that increases the risk by 39%. Levels of cytokines were also elevated in patients with GD, but no association was found between levels of cytokines and the development of GO as there was no change in levels of cytokines.

Conclusions

We suggest that, PARP-1 and NFkB1 gene polymorphisms may be risk factors for developing Graves Disease and Ophthalmopathy.     

Stepwise development of structure–activity relationship of diverse PARP-1 inhibitors through comparative and validated in silico modeling techniques and molecular dynamics simulation

Inhibitors of poly (ADP-ribose) polymerase-1 (PARP-1) enzyme are useful for the treatment of various diseases including cancer. Comparative in silico studies were performed on different ligand-based (2D-QSAR, Kernel-based partial least square (KPLS) analysis, Pharmacophore Search Engine (PHASE) pharmacophore mapping), and structure-based (molecular docking, MM-GBSA analyses, Gaussian-based 3D-QSAR analyses on docked poses) modeling techniques to explore the structure–activity relationship of a diverse set of PARP-1 inhibitors. Two-dimensional (2D)-QSAR highlighted the importance of charge topological index (JGI7), fractional polar surface area (Jurs_FPSA3), and connectivity index (CIC2) along with different molecular fragments. Favorable and unfavorable fingerprints were demonstrated in KPLS analysis, whereas important pharmacophore features (one acceptor, one donor, and two ring aromatic) along with favorable and unfavorable field effects were demonstrated in PHASE-based pharmacophore model. MM-GBSA analyses revealed significance of different polar, non-polar, and solvation energies. Docking-based alignment of ligands was used to perform Gaussian-based 3D-QSAR study that further demonstrated importance of different field effects. Overall, it was found that polar interactions (hydrogen bonding, bridged hydrogen bonding, and pi–cation) play major roles for higher activity. Steric groups increase the total contact surface area but it should have higher fractional polar surface area to adjust solvation energy. Structure-based pharmacophore mapping spotted the positive ionizable feature of ligands as the most important feature for discriminating highly active compounds from inactives. Molecular dynamics simulation, conducted on highly active ligands, described the dynamic behaviors of the protein complexes and supported the interpretations obtained from other modeling analyses. The current study may be useful for designing PARP-1 inhibitors.     

Topoisomerase IIβ associates with Ku70 and PARP-1 during double strand break repair of DNA in neurons

In the present study, the activity of Topoisomerase IIβ (TopoIIβ) is evaluated during peroxide induced double stranded DNA breaks (DSBs) repair in primary neurons. The results showed that the TopoIIβ levels were enhanced during recovery from peroxide mediated damage (PED) along with Ku70, PARP-1, pol beta, and WRN helicase. Furthermore, siRNA mediated knock-down of TopoIIβ in primary neurons conferred enhanced susceptibility to PED in neurons. DSBs in neurons are repaired through two pathways, one promoted by Ku70, while the other is by PARP-1 dependent manner. Participation of TopoIIβ in both pathways was assessed by analysis of the interaction of TopoIIβ with Ku70 and PARP-1 using co-immunoprecipitation experiments in extracts of neurons under peroxide treatment and recovery. The results of these studies showed a strong interaction of TopoIIβ with Ku70 as well as PARP-1 suggesting that TopoIIβ is associated both in Ku70 and PARP-dependent pathways in DSBs repair in primary neurons. The study has thus established that TopoIIβ is an essential component in DSBs repair in primary neurons in both Ku70 and PARP-1 dependent pathways. We suppose that the interaction of TopoIIβ may provide stabilization of the repair complex, which may assist in maintenance of tensional integrity in genomic DNA.     

PARP-1 抑制剂靶向治疗上皮性卵巢癌的研究进展

简介聚二磷酸腺苷核糖聚合酶-1（PARP-1）及其功能和在DNA 损伤修复中的作用,综述PARP-1 抑制剂的作用机制、发展现状以及在上皮性卵巢癌治疗中的应用,并探讨PARP-1 抑制剂靶向治疗上皮性卵巢癌的临床试验失败原因,展望PARP-1 抑制剂的应用前景,提出需对PARP-1 抑制剂在用于治疗上皮性卵巢癌中的耐药机制和选择性展开深入研究。     

Sirtuins与PARP-1在细胞凋亡过程中的相互作用

哺乳动物Sirtuins家族目前共发现7个成员：SIRT1～SIRT7,它们均为NAD+依 赖性且从细菌到人类都保守的一类酶.人们已经对这7种去乙酰化酶进行了亚细胞定位 .目前,对其研究主要集中在对细胞发育相关的重要转录因子如p53、FOXO家族及相关 蛋白的去乙酰化修饰.Sirtuins对许多生理过程有着重要的调节作用,尤其是当发现 它们对寿命延长的调控作用后,Sirtuins引起了人们极大的关注,且都发表在世界顶 级刊物上.聚ADP核糖聚合酶(poly ADP-ribose polymerase, PARP)是一类存在于大多 数真核细胞中的蛋白质翻译后修饰酶,尤其是聚ADP核糖聚合酶1(PARP-1)在细胞内 DNA损伤修复等过程中起着重要作用,该酶同样以NAD+作为催化反应的底物.有研究发 现,Sirtuins家族成员与PARP-1在细胞内某些重要生理过程中存在着相互作用.本文评 述了Sirtuins家族成员、PARP-1的生物学特点,并就其参与哺乳动物细胞凋亡的调控 机制和相关信号通路进行了详细的论述,以期对Sirtuins家族成员、PARP-1生物学功 能及其相互作用的研究提供理论指导.     

PARP-1 inhibitors enhance the chemosensitivity of leukemia cells by attenuating NF-κB pathway activity and DNA damage response induced by Idarubicin

Idarubicin(IDA),an anthracycline antineoplastic drug,is commonly used in the treatment of acute myeloid leu-kemia(AML)with reasonable response rates and clinica...     

PARP-1、AIF在大鼠脊髓损伤后细胞凋亡中的作用

目的探讨聚腺苷二磷酸核糖聚合酶-1(PARP-1)、凋亡诱导因子(AIF)在大鼠脊髓损伤后细胞凋亡中的作用。方法脊髓损伤模型以Allen’s法制备。成年健康SD大鼠随机分为损伤组、PARP-1抑制剂组和假手术组。每组再分1d、3d、7d、14d时间点。苏木精-伊红(HE)染色观察脊髓组织形态学变化;用免疫组织化学方法检测各组各时间点PARP-1、AIF的表达变化;原位末端脱氧核糖核酸转移酶介导dUTP标记法(TUNEL)检测细胞的凋亡水平。结果 HE染色结果显示,与假手术组相比,损伤组脊髓结构破坏明显,大量炎性细胞浸润,许多神经元变性坏死;抑制剂组与损伤组比较,脊髓损伤程度减轻,存活神经元较多。免疫组化结果显示,与假手术组相比,术后1~14d损伤组脊髓PARP-1、AIF表达明显增强(P0.57),且于3d达高峰(P0.05);与损伤组相比,抑制剂组各时间点脊髓PARP-1、AIF阳性表达均显著降低(P0.05)。TUNEL结果显示,与假手术组比较,损伤组阳性细胞凋亡率明显升高,且在损伤后3d最高(均P0.05);而抑制剂组各时间点细胞凋亡率均明显低于损伤组,但高于假手术组(均P0.05)。结论大鼠脊髓损伤后存在PARP-1、AIF介导的细胞凋亡,二者在损伤后细胞凋亡的早期可能起重要作用。     

G6PD缺陷通过Caspase-3和PARP-1诱导人黑色素瘤细胞凋亡

葡糖-6-磷酸脱氢酶(G6PD)在许多肿瘤细胞中高表达,但其发生的作用机理目前仍然不明确.以正常人表皮黑色素细胞(HEM)、野生型人黑色素瘤A375细胞(A375-WT)和G6PD缺陷的A375细胞(A375-G6PDΔ)为对象,经real-time PCR、Western印迹和紫外分光光度法分析显示,A375-WT细胞的mRNA、G6PD蛋白和G6PD活性分别是HEM细胞的1.89倍(P0.05)、6.86倍(P0.01)和2.30倍(P0.05).Annexin V/PI流式细胞仪和Western印迹测定表明,A375-G6PDΔ的凋亡率是A375-WT的5.10倍(P0.01),活化半胱氨酸蛋白酶3(caspase-3)增高1.84倍(P0.01)以及89 kD多聚二磷酸腺苷核糖聚合酶-1(PARP-1)生成增加2.87倍(P0.01).分光光度法分析显示,A375-G6PDΔ的NADPH和GSH分别降低了72.30%(P0.01)和27.39%(P0.05),并伴有75.43%的H2O2增高(P0.01).结果提示,G6PD在黑色素瘤细胞中高表达和高活性,而敲减G6PD表达通过caspase-3和PARP-1信号诱发人黑色素瘤细胞凋亡,这为深入揭示黑色素瘤的发生机理提供了新思路。     

PARP-1沉默对1,4-苯醌致骨髓间充质干细胞微核形成的影响

为探讨聚腺苷二磷酸核糖聚合酶-1(poly(ADP-ribose)polymerase-1,PARP-1)对1,4-苯醌(1,4-benzoquinone,PBQ)诱导骨髓间充质干细胞(bone mesenchymal stem cells,BMSCs)微核形成的影响。我们以空载体BMSCs为对照组,以PARP-1沉默BMSCs为处理组,免疫印迹法检测PARP-1蛋白表达量。磷酸盐缓冲液(PBS)溶解PBQ,分别以0μmol/L、2.5μmol/L、5.0μmol/L、10.0μmol/L、20.0μmol/L、40.0μmol/L、80.0μmol/L、160.0μmol/L和320.0μmol/L PBQ染毒两组细胞,应用MTT法检测细胞活力。根据细胞活力检测结果选择合适染毒浓度后进行微核试验,实时荧光定量-聚合酶链反应检测PARP-1基因表达。结果显示,沉默细胞PARP-1的蛋白表达量较对照组的细胞((1.00±0.03)倍)下降了85%(p0.05)。染毒24 h后,相同剂量下两组细胞的活力并无显著性差异。随着PBQ染毒剂量增加,两组细胞PARP-1表达水平、微核率和核异常率均明显增高(p0.05),且相同剂量下PARP-1沉默组细胞比对照组细胞微核率及核异常率明显增高(p0.05)。结果表明,PARP-1沉默BMSCs在PBQ作用下更易发生DNA损伤,且PARP-1沉默不利于DNA损伤的修复。     

CHFR,PARP-1基因对B细胞淋巴瘤Raji细胞增殖和凋亡的影响（英文）

目的:探讨CHFR与聚(ADP-核糖)聚合酶1(PARP-1)基因对B细胞淋巴瘤Raji细胞增殖和凋亡的影响。方法:用5-Aza-d C处理Raji细胞,后通过qRT-PCR检测CHFR的mRNA表达水平,western blot评估CHFR、PARP-1的蛋白表达。经CHFR慢病毒转染Raji细胞后,用qRT-PCR和western blot评估RAJI细胞中的CHFR、PARP-1变化。通过CCK-8法测定细胞的增殖情况,流式细胞术检测细胞周期的变化。结果:与对照组相比,经5-Aza-d C处理Raji组的CHFR mRNA表达显著上调(P0.01),PARP-1mRNA水平和蛋白表达水平明显降低(P0.05)。与对照组相比,Sh RNA组CHFR的mRNA表达显着下调(均P0.01),PARP-1mRNA和蛋白表达水平升高(P0.05)。CCK结果显示CHFR Sh RNA组细胞活力明显低于对照组(P0.05)。流式细胞术结果显示CHFR沉默后细胞凋亡率降低(P0.05)。结论:CHFR可能通过PARP-1调控B淋巴细胞的增殖和凋亡。     

CHFR, PARP-1基因对B细胞淋巴瘤Raji细胞增殖和凋亡的影响

摘要 目的：探讨CHFR与聚(ADP-核糖)聚合酶1(PARP-1)基因对B细胞淋巴瘤Raji细胞增殖和凋亡的影响。方法：用5-Aza-dC处理Raji细胞,后通过qRT-PCR检测CHFR的mRNA表达水平,western blot评估CHFR、PARP-1的蛋白表达。经CHFR慢病毒转染Raji细胞后,用qRT-PCR和western blot评估RAJI细胞中的CHFR、PARP-1变化。通过CCK-8法测定细胞的增殖情况,流式细胞术检测细胞周期的变化。结果：与对照组相比,经5-Aza-dC处理 Raji组的CHFR mRNA表达显著上调(P <0.01),PARP-1 mRNA水平和蛋白表达水平明显降低(P <0.05)。与对照组相比,ShRNA组CHFR的mRNA表达显着下调(均P <0.01),PARP-1 mRNA和蛋白表达水平升高(P <0.05)。CCK结果显示CHFR ShRNA组细胞活力明显低于对照组(P <0.05)。流式细胞术结果显示CHFR沉默后细胞凋亡率降低(P <0.05)。结论：CHFR可能通过PARP-1调控B淋巴细胞的增殖和凋亡。     

PARP-1 Inhibitor,DPQ, Attenuates LPS-Induced Acute Lung Injury through Inhibiting NF-κB-Mediated Inflammatory Response

Acute lung injury (ALI) is characterized by overwhelming lung inflammation and anti-inflammation treatment is proposed to be a therapeutic strategy for ALI. Poly (ADP-ribose) polymerase-1 has been demonstrated to be involved in tissue inflammation and one of its inhibitors, 3, 4-Dihydro-5[4-(1-piperindinyl)butoxy]-1(2H)-isoquinoline (DPQ), exerts anti-inflammatory effect. However, it is still unclear whether the DPQ possesses the protective effect on ALI and what mechanisms are involved. In this study, we tested the effect of DPQ on the lung inflammation induced by lipopolysaccharide (LPS) challenge in mice. We found that 6 h-LPS challenge induced significant lung inflammation and vascular leakage in mice. Treatment with DPQ at the dose of 10 μg/kg markedly reduced the neutrophil infiltration, myeloperoxidase activity and up-regulation of pro-inflammatory mediators and cytokines. LPS-elevated vascular permeability was decreased by DPQ treatment, accompanied by the inhibition of apoptotic cell death in mice lungs. In addition, we isolated mice peritoneal macrophages and showed pretreatment with DPQ at 10 μM inhibited the production of cytokines in the macrophages following LPS stimulation. DPQ treatment also inhibited the phosphorylation and degradation of IκB-α, subsequently blocked the activation of nuclear factor (NF)-κB induced by LPS in vivo and in vitro. Taken together, our results show that DPQ treatment inhibits NF-κB signaling in macrophages and protects mice against ALI induced by LPS, suggesting inhibition of Poly (ADP-ribose) polymerase-1 may be a potential and effective approach to resolve inflammation for the treatment of ALI.