Association of poly（ADP-ribose） polymerase activity in circulating mononuclear ceils with myocardial dysfunction in patients with septic shock

Background Severe sepsis and septic shock are the leading causes of morbidity and mortality in hospitalized patients.This study aimed to investigate the association of poly（ADP-ribose） polymerase-1 （PARP-1） activity in circulating mononuclear cells with myocardial dysfunction in patients with septic shock.Methods A total of 64 patients with septic shock were divided into the survival group （n=41） and the nonsurvival group （n=23） according to mortality at 28 days after enrollments.PARP-1 activity in circulating mononuclear cells,brain natriuretic peptide,Acute Physiology and Chronic Health Evaluation Ⅱ score,the cardiac index （CI）,the cardiac function index （CFI）,global ejection fraction （GEF）,and the left ventricular contractility index （dp/dt max） were measured after admission to the intensive care unit.Results PARP-1 activity in circulating mononuclear cells of nonsurvival patients with septic shock was significantly higher than that in survival patients.PARP-1 activity in circulating mononuclear cells was strongly,negatively correlated with the CI,the CFI,GEE and dp/dt max.Multiple Logistic regression analysis showed that PARP-1 activity in circulating mononuclear cells was an independent risk factor of myocardial dysfunction.The optimal cutoff point of PARP-1 activity for predicting 28-day mortality was 942 nmol/L with a sensibility of 78.2% and specificity of 65.1%.Conclusion PARP-1 activity in circulating mononuclear cells is significantly associated with myocardial dysfunction and may have prognostic value in patients with septic shock.     

聚ADP核糖聚合酶抑制剂用于卵巢癌治疗的研究进展

卵巢癌是导致女性死亡的主要癌症之一,死亡率在妇科癌症中居高不下.聚ADP核糖聚合酶(PARP)抑制剂可靶向杀灭同源重组修复缺陷的肿瘤细胞,包括乳腺癌易感基因(BRCA)突变的癌细胞,这种机制被称为协同致死效应.目前已有3个PARP抑制剂被FDA批准用于治疗晚期BRCA突变的卵巢癌.本文旨在阐述PARP抑制剂治疗卵巢癌的作用机制,并对已进入临床试验阶段的PARP抑制剂进行综述,以期为开发新的卵巢癌治疗药物提供参考.     

PARP抑制剂与合成致死

随着人们对恶性肿瘤生物学认识的加深,一些新的抗肿瘤策略不断出现。例如之前已有基于肿瘤中的癌基因依赖现象的分子靶向疗法成功用于临床,而合成致死成为当下抗肿瘤药物发展的又一新的方向。研究表明,PARP-1与BRCA1/2之间为合成致死的关系。本文对PARP-1抑制与BRCA1/2缺陷如何构成其合成致死作用以及此作用在抗肿瘤中的潜在应用价值进行综述。     

PARP抑制剂5-AIQ对CT26细胞侵袭及转移的影响

目的探讨多聚(腺苷二磷酸核糖)聚合酶[Poly(ADP-ribose)polymerases,PARPs]抑制剂5-氨基异喹啉酮[5-Aminoisoquinolinone.HCl,5-AIQ]对小鼠结肠腺癌CT26细胞基质黏附、运动、侵袭能力的影响。方法应用Westernblot检测5-AIQ对PARP表达的影响,细胞-基质黏附实验、细胞运动及侵袭实验观察5-AIQ抑制CT26细胞PARP后,其基质黏附、运动及侵袭能力的变化。结果5-AIQ处理组PARP表达明显弱于未处理组(P<0.05);黏附实验结果显示,与5-AIQ未处理组比较,5-AIQ100、500μmol/L处理组的吸光度值明显降低(P<0.01),其黏附抑制率分别是14.54%、21.69%;运动及侵袭实验中,5-AIQ500μmol/L处理组与未处理组的细胞数差别显著(P<0.05),其运动及侵袭抑制率分别是30.09%、37.47%。结论5-AIQ可抑制CT26细胞PARP的表达,并可降低CT26细胞基质黏附、运动及侵袭能力,5-AIQ在抗肿瘤侵袭转移中可能发挥作用。     

多(二磷酸腺苷-核糖)水解酶抑制剂单宁酸对糖尿病大鼠肾脏病变的影响及其机制

目的：研究多（二磷酸腺苷-核糖）水解酶（PARG）抑制剂单宁酸（GLTN）对糖尿病
大鼠肾脏病变的影响并探讨其可能的机制。方法：将实验动物随机分为正常对照组（NC组）、糖尿病组（DM组）、糖尿病3-氨基苯甲酰胺（3-AB）处理组（CDT组）、糖尿病GLTN低剂量（20 mg·kg-1·d^-1）处理组（LDT组）和糖尿病GLTN高剂量（30 mg·kg-1·d^-1）处理组（HDT组）,12周后检查各组动物血糖、血肌酐、尿素氮、尿蛋白排泄率水平及肾脏病变情况,免疫组织化学法检测各组大鼠肾组织PARP阳性信号表达。结果：与DM组比较,LDT和HDT组血糖水平明显降低（P<0.01）,血肌酐、尿素氮水平和尿蛋白排泄率也明显降低(P<0.01,P<0.05,P<0.01）,LDT和HDT组糖尿病大鼠的肾脏病变程度减轻,肾组织PARP蛋白表达降低(P<0.05)。但LDT和HDT组之间上述肾功能参数比较差异无显著性(P>0.05)。结论：GLTN可降低糖尿病大鼠的血糖水平,抑制肾组织PARP表达,改善肾脏功能和形态异常。     

Effect of the regimen of "Gaoshan Hongjingtian" on mechanism of Poly (ADP-ribose) polymerase regulation of nuclear factor kappa B in the experimental diabetic retinopathy

Background Poly (ADP-ribose) polymerase (PARP) plays an important role in the death of retinal capillary cells in diabetic retinopathy (DR), partly via its regulation of nuclear factor kappa B (NF-κB). The current study investigated the effect of the regimen of "Gaoshan Hongjingtian" on the mechanism of PARP regulation of NF-κB, and demonstrated the possible impact of the regimen of "Gaoshan Hongjingtian" and rhodiola sachalinensis on diabetic retinopathy. Methods Wistar rats were made diabetic with streptozotocin, then, were assigned to three groups. After 2 months, these diabetic rats treated with rhodiola sachalinensis or the regimen of “Gaoshan Hongjingtian”, or untreated. Analyses of expression levels of PARP, NF-κB and intercellular adhesion molecule-1 (ICAM-1) on the retinas of rats in different groups were performed by Western blotting and immunohistochemical assays, and mRNA levels of NF-κB and ICAM-1were determined by real-time PCR. In addition, the basement membranes of capillaries in the rats’ retinas were observed by using electron microscopy, and diabetes-induced capillary degeneration (ghost pericytes and acellular capillaries) were quantitated. Results From the third month after the injection, the diabetic rats were given daily with the regimen of “Gaoshan Hongjingtian”, rhodiola sachalinensis or tap water separately. The diabetic rats failed to gain weight when compared with normal age-matched ones, whereas their glycated hemoglobin levels were significantly increased. After 5 months, the mRNA levels of NF-κB and ICAM-1, the protein expression of PARP, NF-κB, and ICAM-1 were significantly increased in the retinas of diabetic rats in untreated group, compared to the nondiabetic controls. After 8 months, the number of degenerated retinal capillaries (ghost pericytes and acellular capillaries) was significantly increased in the diabetic rats in untreated group, compared to normal age-matched rats. The regimen of “Gaoshan Hongjingtian” and rhodiola sachalinensis inhibited diabetes-induced up-expression of PARP, NF-κB, and ICAM-1 in the retinas of diabetic rats with diabetic duration of 5 months. Treatment using the regimen of “Gaoshan Hongjingtian” and rhodiola sachalinensis significangtly inhibited increases in the number of acellular capillaries and pericyte ghosts and suppressed basement membrane thickening in the retinas of rats with diabetes for 8 months, as compared to control diabetic rats. Conclusions These results indicate that PARP plays a role in the pathogenesis of diabetic retinopathy. Rhodiola sachalinensis and the regimen of “Gaoshan Hongjingtian” may have acted on the mechanism of PARP regulation of NF-κB, which suppressed the expression of NF-κB and ICAM-1, and led to inhibition of retinal capillary degeneration.     

多聚腺苷二磷酸核糖聚合酶抑制剂AG014699联合化疗对三阴性乳腺癌细胞株MDA-MB-231增殖的影响

目的 研究多聚腺苷二磷酸核糖聚合酶(PARP)抑制剂AG014699联合多西他赛(DTX)或卡铂(CBP)对三阴性乳腺癌细胞株MDA-MB-231增殖的影响,探讨PARP抑制剂AG014699联合化疗是否有协同抗肿瘤效应。方法 PARP抑制剂AG014699与DTX、CBP单独或联合作用于MDA-MB-231细胞,细胞增殖及细胞毒性实验法检测细胞增殖并用联合用药公式分析合用效应(q值0.85～1.15为单纯相加,>1.15为协同,<0.85为拮抗);流式细胞仪分析细胞凋亡及周期分布。结果 PARP抑制剂AG014699、DTX、CBP单独作用于MDA-MB-231细胞,均可抑制增殖,诱导凋亡,引起细胞周期阻滞;PARP抑制剂AG014699(10 μmol/L)与DTX(10^-8、10^-7、10^-6、10^-5 mol/L)、CBP(10^-5、10^-4 mol/L)联合作用时,q值在0.85～1.15,显示相加效应;PARP抑制剂AG014699与CBP(10^-3 mol/L)联合作用时,q值＞1.15,显示协同效应。PARP抑制剂AG014699联合DTX或CBP能进一步促进凋亡,并使G2/M期细胞比例增加。结论 PARP抑制剂AG014699联合化疗药物DTX或CBP能显著抑制MDA-MB-231细胞增殖,发挥相加或协同抗肿瘤作用。     

抑制PARP1活性对博来霉素诱导的肺纤维化的改善作用

目的:探讨抑制聚ADP核糖多聚酶1(PARP1)活性对于博莱霉素导致的肺纤维化的改善作用,阐明抑制PARP1活化改善肺纤维化的作用机制。方法:48只雌性ICR小鼠随机分为对照组(腹腔注射生理盐水)和5、10、15 mg·kg^-1博莱霉素组(模型组)。3个模型组分别在第1、4、8、11、15、18、22和25天腹腔注射相应浓度的博莱霉素。60只雌性ICR小鼠随机分成对照组、10 mg·kg^-1博莱霉素模型组及不同剂量PARP1抑制剂PJ34治疗组(腹腔注射博来霉素20min前尾静脉注射1、5和10 mg·kg^-1 PJ34)。上述实验中,分别于第14、21和28天每组选取3只小鼠,处死后取肺。HE染色观察肺泡结构,Masson染色分析纤维化程度。肺上皮源性细胞A549用于博莱霉素刺激后,Real-time PCR法分析PJ34对上皮间充质转化(EMT)标志基因表达的影响。结果:与博莱霉素模型组比较,PJ34治疗组小鼠肺泡结构的破坏及胶原在肺间质中的沉积明显减轻;Real-time PCR法分析,与博莱霉素模型组比较,PJ34治疗组肺上皮细胞A549中E-cadherinmRNA表达水平升高(P<0.05),α-SMA和TGF-βmRNA表达水平下降(P<0.05或P<0.01)。结论:抑制PARP1活性可以影响EMT标志基因的表达及肺纤维化的产生,PARP1抑制剂可用于预防博来霉素治疗肿瘤过程中产生的肺纤维化。     

Poly （ADP-ribose） polymerase inhibitor reduces heart ischaemia/ reperfusion injury via inflammation and Akt signalling in rats

Background Poly (ADP-ribose) polymerase (PARP) has been proposed to play an important role in the pathogenesis of heart ischaemia/reperfusion (I/R)injury.3,4-dihydro-5-[4-(1-piperidinyl)butoxy]-1(2H)-i...     

抑制聚二磷酸腺苷核糖聚合酶1减轻尼古丁诱导的支气管上皮细胞炎症反应

目的 通过检测尼古丁刺激的支气管上皮细胞中炎症因子的变化,探讨聚二磷酸腺苷核糖聚合酶1(PARP1)在Toll样受体4(TLR4)介导的炎症中的作用和机制.方法 培养支气管上皮细胞.分别检测尼古丁刺激组及其阴性对照组、TLR4和PARP1抑制组及其阴性对照组中炎症因子相关基因、蛋白的表达量.结果 与对照组比较,尼古丁刺激增加了TLR4和PARP1的蛋白表达.TLR4抑制减少了尼古丁诱导的诱导性一氧化氮合酶(iNOS)、细胞间黏附分子1(ICAM-1)和PARP1的上调.核因子κB的抑制减少了iNOS和ICAM-1的表达.抑制PARP1通过阻止核因子κB的核转位而减少了尼古丁诱导的炎症因子表达.结论 尼古丁刺激通过TLR4/PARP1/NF-κB途径诱导ICAM-1和iNOS的表达.在尼古丁诱导、TLR4介导的炎症反应中,PARP-1或许是不可或缺的因素.在支气管上皮不典型增生中,PARP-1抑制或许是降低轻尼古丁诱导的炎症因子表达的有效手段.     

低剂量氢醌对大鼠骨髓间充质干细胞生物学性状及PARP-1表达的影响

目的：探讨低剂量氢醌(HQ)对大鼠骨髓间充质干细胞（BMSCs）生物学性状及聚腺苷二磷酸核糖聚合酶-1(PARP-1)基因表达的影响,阐明低剂量HQ对骨髓的毒效应。方法：磷酸盐缓冲液(PBS)溶解HQ,以PBS处理组为对照组,分别以2.5、5.0和10.0 μmol•L^-1 HQ染毒大鼠BMSCs为处理组。应用Cell Counting Kit-8试剂盒检测细胞活力和细胞增殖能力,通过磷脂结合蛋白(Annexin V) /碘化丙啶(PI)标记的流式细胞技术检测细胞凋亡,用实时荧光定量-聚合酶链反应(qRT-PCR)检测PARP-1基因表达的改变。结果：与PBS对照组比较,各剂量组细胞增殖能力明显提高(P<0.05);染毒48 h后,5.0和10.0 μmol•L^-1 HQ组细胞的凋亡率均明显降低(P<0.05);染毒48 h后,2.5、5.0和10.0 μmol•L^-1 HQ组细胞PARP-1表达量分别为对照组0.92、0.56(P<0.05)和0.45倍(P<0.05)。结论：低剂量HQ能抑制大鼠BMSCs凋亡和PARP-1基因表达,促进细胞增殖。     

多聚二磷酸腺苷(ADP)核糖聚合酶-1介导高糖致大鼠肾脏系膜细胞tPA/PAI-1的紊乱

目的 探讨多聚二磷酸腺苷(ADP)核糖聚合酶-1(PARP-1)在高糖(25 mmol/L)作用下大鼠肾脏系膜细胞tPA/PAI-1纤溶系统紊乱中的作用.方法 (1)体外培养大鼠肾脏系膜细胞株(MCs 1097),使用高糖(25 mmol/L)处理大鼠肾脏系膜细胞,部分实验组中应用PARP-1特异性抑制剂PJ34(3×10-6 mol/L)进行干预处理.(2) RT-PCR及Western blot检测PARP-1的mRNA及蛋白表达.(3)高通量比色测定法检测PARP-1的活性.(4) ELISA法测定细胞上清液tPA、PAI-1的分泌蛋白.结果 (1)高糖显著诱导大鼠肾脏系膜细胞PARP-1mRNA和蛋白的表达,PJ-34可明显抑制高糖诱导的PARP-1 mRNA和蛋白的过度表达.同时,高糖显著诱导大鼠肾脏系膜细胞PARP-1激活,PJ-34可显著抑制高糖诱导的PARP激活.(2)高糖轻度减少大鼠肾脏系膜细胞分泌型tPA的蛋白表达,显著增加大鼠肾脏系膜细胞分泌型PAI-1的蛋白表达,导致tPA/PAI-1比值降低,PJ-34显著预防高糖诱导的上述改变.结论 高糖可以诱导培养的大鼠肾脏系膜细胞内PARP激活,PARP1表达增加,PJ-34预处理可以明显降低高糖诱导的大鼠肾脏系膜细胞内PARP的激活以及下游tPA/PAI-1纤溶系统紊乱,提示高糖可能通过过度激活PARP来诱导tPA/PAI-1功能紊乱.     

Shh-PARP-1信号通路在茶多酚拮抗胰岛微血管内皮细胞脂毒性中的调控作用

目的 探讨Sonic Hedgehog (Shh)-聚腙苷二磷酸核糖聚合酶1[poly(ADP-ribose) polymerase 1,PARP-1]信号通路在茶多酚拮抗胰岛微血管内皮细胞脂毒性中的调控作用.方法 以小鼠胰岛微血管内皮MS-1细胞为研究对象,分为正常对照组、溶剂对照组、脂肪酸(0.25 mmol/L软脂酸+0.5 mmol/L油酸)组、茶多酚(25μmol/L)组、脂肪酸十茶多酚组、PARP-1抑制剂(8μmol/L BYK204165)+脂肪酸组、PARP-1抑制剂十脂肪酸十茶多酚组、Shh抑制剂(2.5μmol/L环巴胺)+脂肪酸组、Shh抑制剂十脂肪酸十茶多酚组及Shh抑制剂+PARP-1抑制剂+脂肪酸+茶多酚组,分别检测各组细胞活力、凋亡水平、一氧化氮(NO)合成及氧化应激相关指标的改变.结果 脂肪酸处理后,MS-1细胞存活率下降,细胞凋亡率增高(P＜0.05);同时,细胞内NO的含量及总一氧化氮合酶(tNOS)、诱导型NOS(iNOS)和结构型NOS(cNOS)的活性均升高(P＜0.05);而且,脂质过氧化产物丙二醛(MDA)含量增加(P＜0.05),抗氧化物质谷胱甘肽(GSH)和超氧化物歧化酶(SOD)的水平下降(P＜0.05),并增强了PARP-1和磷酸化Shh的表达水平(P＜0.05).茶多酚干预后,各项指标的水平均得以改善(P＜0.05);而且,利用BYK204165和环巴胺预处理1h后,茶多酚对脂肪酸的拮抗效应更为显著,各项检测指标与正常对照组比较差异无统计学意义(P＞0.05).结论 脂肪酸可诱发胰岛微血管内皮功能损伤,茶多酚具有拮抗脂肪酸毒性的作用,且抑制Shh-PARP-1信号通路能增强茶多酚的保护效应.     

大鼠肠缺血再灌注损伤中聚腺苷二磷酸核糖聚合酶-1活性变化的实验研究

目的 探讨肠缺血再灌注损伤中聚腺苷二磷酸核糖聚合酶-1[poly(ADP-ribose)polymerase-1,PARP-1]活性变化情况.方法 通过手术方法阻断腹腔干和肠系膜上动脉45 min后恢复血流,建立大鼠肠缺血再灌注模型为I/R组(n=10),假手术组为Sham组(n=10).HE染色观察肠缺血再灌注后组织学改变;PARP-1的活化程度用免疫组织化学方法检测其产物聚腺苷二磷酸核糖[poly(ADP-ribose),PAR]的表达情况来表示;用Western Blot检测PARP-1的裂解片段p89,探测有无凋亡早期信号表达.结果 I/R组肠黏膜明显受损[病理损伤评分I/R组(14.2±2.6)分vs Sham组(2.5±1.1)分,P＜0.05];仅I/R组可见凋亡早期信号p89表达;与Sham组相比PAR呈显著阳性表达[I/R组(40.5±10.2)%vs Sham组(5.3±3.4)%,P＜0.05].结论 肠缺血再灌注过程中PARP-1被过度激活,可能在导致肠损伤中发挥重要作用.     

Effect of the regimen of Gaoshan Hongjingtian on the mechanism of poly（ADP-ribose） polymerase regulation of nuclear factor kappa B in the experimental diabetic retinopathy

Background Poly(ADP-ribose) polymerase (PARP) plays an important role in the death of retinal capillary cells indiabetic retinopathy (DR) partly via its regulation of nuclear factor kappa B (NF-κB).The...     

Effects of 3-aminobenzamide on expressions of poly（ADP ribose） polymerase and apoptosis inducing factor in cardiomyocytes of rats with acute myocardial infarction

Background Poly（ADP-ribose） polymerase （PARP） plays an important role in cell survival and death. However, the mechanisms involved are not fully understood. Therefore, we investigated the effect of inhibition of PARP on acute myocardial infarction （AMI） at different time points in rats. Methods AMI was induced in rats by ligating the left anterior descending coronary artery. One group received 3-aminobenzamide （3-AB, a kind of PARP inhibitor） （30 mg/kg） by intraperitoneal injection. The changes of ultramicrostructure of cardiocytes in infarction region were noted, PARP cleavage was measured by Western blotting, and expressions of protein of PARP and apoptosis inducing factor （AIF） were measured by immunohistochemical staining after treatment with 3-AB for 2 hours, 4 hours, 6 hours, 1 week, 4 weeks and 8 weeks. Results Few damages to the ultramicrostructure of cardiocytes were observed after treatment with 3-AB. PARP cleavage was detected as early as 4 hours and markedly increased by 6 hours following AMI without 3-AB, but was not found until 6 hours following AMI treated with 3-AB. There were significant differences between 3-AB and AMI groups at the same time points. The expression of PARP was observed gradually increased, but that of AIF was suppressed for 6 hours after treatment of 3-AB, compared with AMI groups in positive cells at the same time points. There was significantly less cleavage of PARP and more PARP expression in 3-AB treated group compared with AMI and control groups at all matched time points. Conclusions Our results suggest that 3-AB inhibits degradation of PARP, increases the expression of PARP protein, and suppresses the expression of AIF protein. Inhibition of PARP activity may protect cardiocytes in rats with AMI and reduce apoptosis.     

大鼠急性心肌梗死后缺血缺氧心肌早期凋亡信号分子的变化

目的探索能够应用于临床辨别早期缺血心肌生物学活性的敏感代谢变化指标。方法SD大鼠行冠状动脉前降支结扎术建立心肌缺血模型,分别结扎0（对照组）、5、20、45min。采用luciferin/luciferase法,RT-PCR法和免疫组织化学法分析心肌组织在不同缺血时间段梗死区、梗死边缘区及正常区ATP含量变化,葡萄糖调节蛋白75（grp75）和缺氧诱导因子1α（HIF1α）基因表达变化,以及细胞色素C释放、聚腺苷二磷酸核糖聚合酶（PARP）降解的情况。结果大鼠冠状动脉前降支分别结扎5、20、45min时,梗死区及梗死区边缘心ATP含量上升,并于20、45min时明显高于正常水平;grp75及HIF1α基因转录水平未见明显改变。免疫组化结果显示少数细胞细胞色素C的释放出现于冠状动脉前降支结扎5min,而PARP的降解发生较迟,出现于冠状动脉前降支结扎20min。冠状动脉前降支结扎45min时细胞色素C的释放和PARP的降解明显增强免疫反应呈现片状染色。结论大鼠急性心肌缺血后早期梗死区发生细胞凋亡,细胞色素C释放、PARP降解出现较早,可用作为临床辨别缺血心肌生物学活性的早期变化指标。     

中国汉族、布依族和壮族群体中聚腺苷二磷酸核糖聚合酶-1基因多态性

【目的】探讨人类聚腺苷二磷酸核糖聚合酶-1(PARP-1)5个外显子核苷酸多态性。【方法】采用聚合酶链式反应-单链构象多态性(PCR-SSCP)和银染技术检测3个民族(汉族、布依族和壮族)634名正常人PARP-1基因多态性。【结果】在3个民族634名正常人血标本的5个外显子扩增产物SSCP电泳条带中,219名汉族人PARP-1基因5个外显子均未检出多态性条带;203名布依族和212名壮族人PARP-1基因的5个外显子扩增产物中分别有2例的外显子20检出1条多态性条带,其余4个外显子扩增产物未见多态性条带。【结论】PARP-1基因第20外显子上可能存在多态性。     

PTMSP与PMP膜渗透汽化分离性能的研究比较

以聚4－甲基戊烯－1（PMP）为膜材质、分别以环己烷、三氯乙烯以及环己烷／三氯乙烯为溶剂,以浇铸法制备了PMP的均质致密膜。研究了不同溶剂体系的相对溶解能力和挥发速度对PMP膜结晶度和形态结构的影响,并对成膜的渗透汽化特性的影响进行了研究。