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AIM:To explore the synergistic sensitization effect of human umbilical cord mesenchymal stem cell culture supernatant(hUMSC-CM)combined with temozolomide(TMZ)on various glioma cell lines,and to elucidate the underlying mechanisms.METHODS:The hUMSC-CM was harvested using two different serum deprivation tech-niques at 24 and 48 h,and was converted into freeze-dried powder,which was then given to rat malignant glioma cell line RG-2,human astrocytoma cell line U251 and human glioblastoma cell line LN-428 at 5 concentrations(0,1,3,6 and 9 g/L).The effectiveness and sensitivity of hUMSC-CM for inhibiting growth of glioma cells at 24,48 and 72 h were as-sessed using CCK-8 assay.Hematoxylin-eosin(HE)staining combined with CCK-8 assay was employed to evaluate the chemotherapy sensitivity of glioma cells after 48 h of treatment with TMZ at 6 concentrations(0,25,50,100,200 and 400 μmol/L).Two concentrations(3 and 9 g/L)of hUMSC-CM and 3 concentrations(50,100 and 200 μmol/L)of TMZ were chosen for concurrent treatment of glioma cells to assess the proliferation and pathological alterations.TUNEL staining was utilized to detect apoptosis.Flow cytometry was utilized to analyze cell cycle modifications.The expression alterations of apoptosis-inducing proteins,cleaved caspase-3,cleaved caspase-8 and cleaved PARP1,as well as autophagy-inducing proteins beclin-1 and LC3,were examined using Western blot to investigate the synergistic sensitization mechanism of hUMSC-CM combined with TMZ in vitro.RESULTS:The susceptibility of glioma cell lines to hUMSC-CM and TMZ varied,with RG-2 showing the highest sensitivity,followed by U251,and then LN-428.The inhibitory effect of hUMSC-CM(3 and 9 g/L)and TMZ(50,100 and 200 μmol/L)combined treatment on glioma cells was significantly greater than that that of single-agent treatments(P<0.05),demonstrating a dose-and concentration-dependent enhancement.Notably,the combination of 9 g/L hUMSC-CM(C9)with 50 μmol/L TMZ(T50)effectively suppressed glioma cell growth.CCK-8 as-say indicated a significant reduction of cell viability in C9+T50 group compared with either C9 or T50 alone(P<0.05).HE staining and TUNEL staining revealed pronounced morphological changes and significant apoptotic features in glioma cells treated with C9+T50.Flow cytometric analysis confirmed that C9+T50 induced cell cycle arrest in glioma cells.Fur-thermore,compared with control group,the levels of cleaved caspase-3,cleaved caspase-8,cleaved PARP1,beclin-1,and LC3-Ⅱ/LC3-Ⅰ were significantly elevated in the C9+T50-treated glioma cells(P<0.01).CONCLUSION:(1)The concomitant administration of hUMSC-CM and TMZ exerts a broad inhibitory effect on glioma cells,with a synergistic sen-sitization observed across different cell lines.(2)The enhancement of glioma cell sensitivity to TMZ by hUMSC-CM may be attributed to the modulation of caspase-8/caspase-3/PARP1 signaling pathway and the induction of both apoptosis and autophagy in glioma cells.
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Objective To study the effect of formononetin on the cell damage of glucose/oxygen deprivation/reoxy-genation glyconeurons via the PARP1 signaling pathway,and to offer theoretical support for the use of Caragana isoflavones in the treatment of cerebral ischemia-reperfusion injury.Methods In mouse neurons(HT22),a model of Oxygen-glucose deprivation/reoxygenation(OGD/R)was created.Western blot was used to detect the expres-sion of PARP1 and PARG in HT22 neurons at various time points of glucose-oxygen deprivation/reoxygenation,and the optimal time point of pathway modification was chosen.After OGD/R,HT22 cells were treated with form-ononetin,PARP1 inhibitor(PJ34),and PARG inhibitor,and six groups were developed:control group,control group+formononetin group,OGD/R group,OGD/R+formononetin group,OGD/R+PJ34 group,OGD/R+PARG inhibitor group.HT22 cells were grown normally without OGD/R therapy in the control group.The expres-sion levels of apoptotic factors and associated proteins in each group were determined using immunofluorescence and Western blot.Results PARP1 pathway was activated most obviously in HT22 cells after 3 hours of glucose and ox-ygen deprivation/reoxygenation.Under the condition of OGD/R 3 h,treatment with formononetin,PJ34 or PARG inhibitor could increase E3 ubiquitin ligase(Iduna),inhibit the expression of PARP1 and PARG pathway proteins,reduce the expression of AIF and P53,and increase the phosphorylation level of AKT protein.Conclusion Form-ononetin can block the PARP1/AIF/Akt signaling pathway by raising the expression of Iduna protein in the pres-ence of OGD/R,hence decreasing the damage to HT22 mouse neurons.
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Objective:To establish an experimental model of chronic obstructive pulmonary disease(COPD)in rats,and explore whether polyadenosine diphosphate ribose polymerase-1(PARP-1)inhibitors regulate Sirtuin 1(SIRT1)and peroxisome proliferator-activated receptor-γ co-activator 1α(PGC-1α)to reduce inflammation and oxidative stress in COPD rats,and explore the possibility of SIRT1-PGC-1α axis as a new target of PARP-1 inhibitor.Methods:Twelve of 48 SD rats were randomly selected as healthy group,and the remaining rats were used to construct experimental models of COPD.Rats that were successfully modeled were randomly divided into model group,PARP-1 inhibitor treatment group and PARP-1 inhibitor+PGC-1α inhibitor group.HE staining was used to observe pathological changes of lung tissue,ELISA was used to detect levels of TNF-α,IL-6,IL-1β,malondialdehyde(MDA),superoxide dismutase(SOD)in rats lung tissue,fluorescence quantitative PCR was used to detect expression levels of SIRT1 and PGC-1α mRNA of rats in each group,Western blot was used to detect expressions of SIRT1 and PGC-1α proteins.Results:Lung tissue structure of rats in healthy group was complete,compared with healthy group,lung tissue of model group suffered structural damage,with a large number of inflammatory cells infiltrated,contents of TNF-α,IL-1β and IL-6 in alveolar lavage fluid were significantly increased,con-tent of MDA in serum was significantly increased,while content of SOD was significantly reduced,and expressions of SIRT1,PGC-1α mRNA and protein were significantly reduced;compared with model group,lung tissue structure of rats in PARP-1 inhibitor treatment group was recovered,and inflammatory cells were reduced,contents of TNF-α,IL-1β and IL-6 were significantly reduced,content of MDA in serum was significantly reduced,while content of SOD was significantly increased,and expressions of SIRT1,PGC-1α mRNA and protein were significantly increased;compared with PARP-1 inhibitor treatment group,the number of inflammatory cells in PARP-1 inhibitor+PGC-1α inhibitor group was increased,contents of TNF-α,IL-1β and IL-6 were significantly increased,content of MDA in serum was significantly increased,while content of SOD was significantly reduced,expressions of SIRT1,PGC-1α mRNA and protein were significantly reduced.Conclusion:PARP-1 inhibitors can alleviate inflammation and oxidative stress by activating SIRT1-PGC-1α axis,thereby effectively alleviating COPD.
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@#Parthanatos is a form of programmed cell death,which is also known as poly(ADP-ribose)polymerase 1(PARP1)-mediated apoptosis-inducing factor(AIF)and macrophage migration inhibitory factor(MIF)-dependent cell death according to its molecular mechanism. Parthanatos is the main cause of a variety of neurodegenerative diseases,such as Parkinson's disease(PD),Alzheimer's disease(AD),motor neuron disease,and is also involved in the pathogenesis of some tumors,such as lung cancer and breast cancer. Therefore,a thorough understanding of the molecular mechanism of Parthanatos is crucial for the therapeutic strategies of related diseases. In recent years,studies have found that effective regulation of the occurrence of Parthanatos by regulating the key proteins PARP1,AIF and MIF is expected to become a therapeutic target for many diseases. Based on the specific molecular mechanism of Parthanatos,this paper reviewed the research progress of therapeutic strategies for related diseases from the aspects of inhibiting and promoting Parthanatos.
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ObjectiveTo investigate whether the effects of paeonol (Pae) on angiotensin Ⅱ (AngⅡ)-induced senescence in vascular smooth muscle cells (VSMCs) were related to angiotensinogen of silencing regulatory information factor 6 (SIRT6)/adenosine diphosphate ribose polymerase 1 (PARP1) signaling pathway in VSMCs. MethodThe model of VSMC-stress aging induced by AngⅡ (100 nmol·L-1) was established. The rats were divided into normal group, model group, low, medium, and high-concentration Pae groups (30, 60, 120 μmol·L-1). The positive rate of cell senescence was detected by SA-β-Gal staining, the ability of cell proliferation was detected by the cell counting kit-8 (CCK-8) method, the expression of SIRT6, PARP1, p16, p21, p53, proliferating cell nuclear antigen (PCNA), deoxyribonucleic acid (DNA)-damaged protein γ-H2AX was detected by Western blot, and VSMC proliferation was detected by EdU staining. The silenced VSMCs were prepared by siRNA-SIRT6 transfection, and the protein expressions of SIRT6, PARP1, p16, and γ-H2AX in VSMCs silenced by SIRT6 were observed. ResultThe results of SA-β-Gal staining showed that the senescence positive rate of SA-β-Gal staining in the model group was higher than that in the normal group (P<0.01), and the positive rate of SA-β-Gal staining in the Pae group was significantly lower than that in the model group (P<0.05, P<0.01). The results of Western blot showed that as compared with the normal group, the expression of PCNA, SIRT6, and PARP1 in the model group was down-regulated, and the expression of aging-related proteins p16, p21, p53, and γ-H2AX was up-regulated in the model group (P<0.05, P<0.01). Compared with the model group, Pae promoted the protein expression of PCNA, SIRT6, and PARP1 and inhibited the protein expression of p16, p21, p53, and γ-H2AX in a dose-dependent manner (P<0.05, P<0.01). The results of EdU staining showed that the number of EdU positive cells in the model group was lower than that in the normal group (P<0.01), and the number of EdU positive cells in Pae groups was significantly higher than that in the model group (P<0.05, P<0.01). After SIRT6 silencing, the effects of Pae on promoting SIRT6 and PARP1 and inhibiting P16 were reversed (P<0.05, P<0.01). In addition, the addition of SIRT6 inhibitor (IN-1) promoted the occurrence of cell senescence induced by AngⅡ (P<0.05, P<0.01). ConclusionPae can effectively inhibit the aging of VSMCs, and its mechanism may be related to the regulation of SIRT6/PARP1 signal pathway.
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@#Phosphatidylinositol-3-kinase (PI3K) inhibitors can increase the sensitivity of tumor cells to Poly ADP-ribose polymerase-1 (PARP-1) inhibitors. Therefore, the simultaneous inhibition of the PARP-1 and PI3K activities are expected to overcome the drug resistance of PARP-1 inhibitors.In our previous work, two compounds XW-1 and WZ-1 with excellent activities against PARP-1 and PI3K were obtained with the limitation to further study due to their poor water solubility.Therefore, XW-1 and WZ-1 were chosen as lead compounds to optimize their solubility by introducing a salt-forming site via a urea group, and 11 novel compounds were designed and synthesized. The structure of all target compounds was confirmed by 1H NMR, 13C NMR, and HRMS.The enzyme activities of the compounds against PARP-1 and PI3K were measured, and the results showed that most of the compounds demonstrated good inhibitory activities against PARP-1 and PI3K.Based on the above result, the inhibitory activities of compounds 8b, 8e, and 8f against MDA-MB-231, MDA-MB-468, HCC1937, HCT116, and olaparib-resistant HCT116R were determined by MTT, respectively.Additionally, the structure-activity relationship was discussed. The results showed that these compounds displayed excellent antiproliferation activity.Among them, compound 8f demonstrated antiproliferation remarkably against all five tumor cells, which was more potent than that of olaparib, and was comparable to that of BKM120.Furthermore, the solubility of hydrochloride salts of compound 8b and 8f was significantly improved compared to the lead compounds.The results of this study will provide a theoretical basis for the further development of PARP-1 and PI3K dual-target inhibitors with good pharmaceutical properties and strong inhibitory activities.
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Increased oxidative stress leads to cell death by inducing DNA damage, PARP activation and energy depletion in age related disorders which are a growing concern due to increased life expectancy. Indeed, cellular NAD+ levels, depletion of which is one of the consequences of overactive PARP, also decline with age. We previously showed rescue in oxidative stress induced paraptotic and necrotic cell death by PARP1 inhibition in D. discoideum. Inhibition of PARP1 activity prevented cellular depletion of its substrate NAD+. To understand the significance of NAD+ depletion in PARP1 mediated oxidative stress induced cell death, exogenous addition of NAD+ was done. Addition of NAD+ prevented PARP1 mediated oxidative stress induced cell death at low doses upto 10 mM NAD+, nevertheless led to an anticipated increase in PARP1 activity. NAD+ significantly prevented oxidative stress induced cell death in D. discoideum. Exogenous NAD+ averted depletion of cellular NAD+ and mitochondrial membrane potential changes that were triggered by oxidative stress, without getting affected by the elevated ROS levels. Altogether, this study ascertains that NAD+ replenishment overcomes cadmium or H2O2 induced cell death by preventing cellular energy collapse incited by PARP1 activation. Thus, our results explicitly demonstrate that PARP1 overactivation led NAD+ depletion but not PARP1 activity per se is of consequential significance in causing oxidative stress induced D. discoideum cell death. Moreover, NAD+ supplementation could be a beneficial approach in aging and age-related disorders mediated by PARP1
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OBJECTIVE@#To investigate the regulatory effect and mechanism of DNA methyltransferase 3A (DNMT3a) in hydroquinone-induced hematopoietic stem cell toxicity.@*METHODS@#Cells (HSPC-1) were divided into 4 groups, that is A: normal HSPC-1; B: HQ-intervented HSPC-1; C: group B + pcDNA3 empty vector; D: group B + pcDNA3- DNMT3a. RT-qPCR and Western blot were used to detect the expression levels of DNMT3a and PARP-1 mRNA and protein, respectively. Cell morphology was observe; Cell viability and apoptosis rate of HSPC-1 were detected by MTT and flow cytometry, respectively.@*RESULTS@#Compared with group A, the expression levels of DNMT3a mRNA and protein in HSPC-1 of group B were decreased, while PARP-1 mRNA and protein were increased (P<0.05); there was no significant difference in the above indexes between group C and group B; compared with group B, the expression levels of DNMT3a mRNA and protein showed increased, while PARP-1 mRNA and protein were decreased significantly in cells of group D transfected with DNMT3a (P<0.05). Cells in each group were transfected with DNMT3a and cultured for 24 h, HSPC-1 in group A showed high density growth and mononuclear fusion growth, while the number of HSPC-1 in group B and C decreased and grew slowly. Compared with group B and C, the cell growth rate of group D was accelerated. The MTT analysis showed that cell viability of HSPC-1 in group B were lower than that of group A at 24 h, 48 h and 72 h (P<0.05); after transfected with DNMT3a, the cell viability of HSPC-1 in group D were higher than that of group B at 24 h, 48 h and 72 h (P<0.05). The apoptosis rate of cells in group B was significantly higher than that of group A (P<0.001), while the apoptosis rate in group D was lower than that of group B (P<0.001).@*CONCLUSION@#DNMT3a may be involved in the damage of hematopoietic stem cells induced by hydroquinone, which may be related to the regulation of PARP-1 activity by hydroquinone-inhibited DNMT3a.
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Humanos , Apoptosis , Proliferación Celular , ADN Metiltransferasa 3A , Células Madre Hematopoyéticas/efectos de los fármacos , Hidroquinonas/toxicidad , Poli(ADP-Ribosa) Polimerasa-1 , ARN Mensajero/metabolismoRESUMEN
Objective:To investigate the effects of naringin on early brain injury in rats with subarachnoid hemorrhage and its possible mechanism of action.Methods:Rats were randomly divided into the sham operation group, the model group, and the naringin group. Each group had 8 rats. The SAH model was established by intravascular perforation, and then rats in the model group and the naringin group were administered 0.9% NaCl or naringin 40 mg/kg by intraperitoneal injection after 0.5 h. SAH score, neurological function score, cerebral edema, and blood-brain barrier permeability were detected. The level of NAD + and nflammatory factors were detected by ELISA. The expression of poly(ADP-ribose) polymerase-1 (PARP-1), apoptosis inducing factor (AIF), and protease-activated receptor (PAR) proteins was detected by Western Blot. The expression of PARP-1 mRNA was detected by quantitative real-time fluorescence PCR (qRT-PCR). Neuronal apoptosis was detected by an immunofluorescence assay. Results:Compared with the model group, naringin treatment improved neurological function ( P<0.01), reduced cerebral edema and Evans blue exudation (all P<0.01), increased the content of NAD + ( P<0.001), reduced IL-1β, IL-6 and TNF-α levels (all P<0.001), and reduced the expression of PARP-1/AIF pathway-related proteins in vivo (all P<0.001). In addition, naringin could inhibit neuronal apoptosis in early brain injury after SAH. Conclusions:Naringin can improve the early brain injury after SAH, which may be achieved by inhibiting the PARP-1/AIF pathway.
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Objective: To observe the platinum drugs resistance effect of N-acetyltransferase 10 (NAT10) overexpression in breast cancer cell line and elucidate the underlining mechanisms. Methods: The experiment was divided into wild-type (MCF-7 wild-type cells without any treatment) group, NAT10 overexpression group (H-NAT10 plasmid transfected into MCF-7 cells) and NAT10 knockdown group (SH-NAT10 plasmid transfected into MCF-7 cells). The invasion was detected by Transwell array, the interaction between NAT10 and PARP1 was detected by co-immunoprecipitation. The impact of NAT10 overexpression or knockdown on the acetylation level of PARP1 and its half-life was also determined. Immunostaining and IP array were used to detect the recruitment of DNA damage repair protein by acetylated PARP1. Flow cytometry was used to detect the cell apoptosis. Results: Transwell invasion assay showed that the number of cell invasion was 483.00±46.90 in the NAT10 overexpression group, 469.00±40.50 in the NAT10 knockdown group, and 445.00±35.50 in the MCF-7 wild-type cells, and the differences were not statistically significant (P>0.05). In the presence of 10 μmol/L oxaliplatin, the number of cell invasion was 502.00±45.60 in the NAT10 overexpression group and 105.00±20.50 in the NAT10 knockdown group, both statistically significant (P<0.05) compared with 219.00±31.50 in wild-type cells. In the presence of 10 μmol/L oxaliplatin, NAT10 overexpression enhanced the binding of PARP1 to NAT10 compared with wild-type cells, whereas the use of the NAT10 inhibitor Remodelin inhibited the mutual binding of the two. Overexpression of NAT10 induced PARP1 acetylation followed by increased PARP1 binding to XRCC1, and knockdown of NAT10 expression reduced PARP1 binding to XRCC1. Overexpression of NAT10 enhanced PARP1 binding to LIG3, while knockdown of NAT10 expression decreased PARP1 binding to LIG3. In 10 μmol/L oxaliplatin-treated cells, the γH2AX expression level was 0.38±0.02 in NAT10 overexpressing cells and 1.36±0.15 in NAT10 knockdown cells, both statistically significant (P<0.05) compared with 1.00±0.00 in wild-type cells. In 10 μmol/L oxaliplatin treated cells, the apoptosis rate was (6.54±0.68)% in the NAT10 overexpression group and (12.98±2.54)% in the NAT10 knockdown group, both of which were statistically significant (P<0.05) compared with (9.67±0.37)% in wild-type cells. Conclusion: NAT10 overexpression enhances the binding of NAT10 to PARP1 and promotes the acetylation of PARP1, which in turn prolongs the half-life of PARP1, thus enhancing PARP1 recruitment of DNA damage repair related proteins to the damage sites, promoting DNA damage repair and ultimately the survival of breast cancer cells.
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Femenino , Humanos , Neoplasias de la Mama/enzimología , Línea Celular Tumoral , Resistencia a Antineoplásicos , Células MCF-7 , Acetiltransferasas N-Terminal/metabolismo , Compuestos Organoplatinos/farmacología , Oxaliplatino/farmacología , Proteína 1 de Reparación por Escisión del Grupo de Complementación Cruzada de las Lesiones por Rayos XRESUMEN
【Objective】 To investigate the effects of resveratrol on gemcitabine chemotherapy in pancreatic cancer and the possible molecular mechanism. 【Methods】 Gemcitabine resistant cell lines were screened by continuous low concentration increasing induction. High-throughput RNA-seq was used to analyze the differential expression enrichment pathway, COMET assay was used to detect DNA damage, Western blotting was used to detect related pathway indicators, and Chou-Talalay was used to calculate drug combination synergistic index. AutoDock predicts docking targets for small molecules and proteins. 【Results】 DNA damage repair related pathways were activated in drug-resistant cell lines compared with their parents. Resveratrol enhanced the DNA damage effects induced by gemcitabine (P<0.01). Resveratrol inhibited the expression of PARP1, a key molecule of DNA damage repair, and played a synergic effect with gemcitabine (CI<1). Resveratrol has docking targets with the CAT domain of PARP1. 【Conclusion】 Resveratrol can inhibit PARP1, a key molecule of chemotherapy resistance, and has a synergistic effect with gemcitabine in pancreatic cancer chemotherapy.
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OBJECTIVE To investigate the pharmacological effect and mechanism of Sanguisorba officinalis L. (SOL) in non-small cell lung cancer (NSCLC) in vitro and in vivo based on network pharmacology. METHODS Network pharmacology was used to analyze the improving effect of SOL on NSCLC and possible targets. Cell counting kit 8 (CCK-8) and 5-ethynyl-2'-deoxyuridine (EdU) staining, Western blotting, flow cytometry of Annexin Ⅴ/PI, Hoechst 33342/PI staining detection and immunofluorescence were utilized in vitro. H&E staining, immunohistochemistry staining and Western blotting were performed in vivo. RESULTS Based on network prediction, we analyzed the 208 common targets of SOL and NSCLC. 36 core targets in 208 common targets were obtained through cytoscape analysis. And the top 10 core targets included Akt, mTOR, EGFR, etc.. KEGG analysis showed that PI3K-Akt signaling pathway was the most likely pathway. Furthermore, the mechanism study found that SOL could activate the PI3K/Akt/mTOR signaling pathway in vitro and in vivo. The anti-proliferative effect of SOL in A549 and H1299 cells was measured and validated by CCK-8 and EdU assay. Immunohistochemical results of Ki67 showed that SOL effectively inhibited tumor growth in vivo. SOL also significantly inhibited the migration and invasion of A549 and H1299 cells. SOL significantly increased the percentage of cells with PI signal in A549 and H1299, and the process of cell death of A549 cells indicated that SOL induced apoptosis. The PARP-1 and caspase-3 in A549 and H1299 were found to be activated in a dose manner. The results in vivo were consistent with those in vitro. CONCLUSION SOL-induced, caspase-3-mediated apoptosis via the induction of the PI3K/Akt/mTOR signaling pathway in NSCLC, which further clarified the mechanism of SOL in the inhibition of NSCLC, and thereby provided a possibility for SOL to serve as a novel therapeutic agent for NSCLC in the future.
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Objective:To explore the mechanism of potassium iodide-induced pyrolysis of thyroid follicular cells.Methods:Thyroid gland tissue was obtained from patients with thyroid cancer (TC) coexisting with Hashimoto′s thyroiditis, and the tumor-adjacent Hashimoto′s thyroiditis tissue was used as the control. ELISA was used to detect the concentration of the pyroptosis inflammatory cytokines interleukin (IL)-1β and IL-18 in the tissues, and Western blotting was used to detect the activation of gasdermin (GSDM) proteins, a biomarker for pyroptosis. Thyroid follicular cells treated with different concentrations of potassium iodide, and IL-1β, IL-18, lactate dehydrogenase (LDH), GSDMD were measured. Transcriptome chip analysis was used to explore the differentially expressed genes involved in pyroptosis of thyroid follicular cells induced by potassium iodide treatment.Results:The levels of IL-1β and IL-18 cytokines in the tissues of patients with Hashimoto′s thyroiditis and thyroid cancer were higher than control tissues ( P<0.01), and the activation of the pyroptosis executive protein GSDMD was significant increased, while GSDME was not activated. IL-1β, IL-18, and LDH secretion were significantly increased in response to potassium iodide stimulation in thyroid follicular cells ( P<0.01) and GSDMD was cleaved, which indicated that potassium iodide induced the pyroptosis of thyroid follicular cells. Moreover, potassium iodide could activate NLRP3 inflammasomes to promotethe pyroptosis of thyroid follicular cells. Transcriptome chip analysis further found that PARP1 protein was highly upregulated by the stimulation of potassium iodide, and then enhanced the activity of nuclear factor-κB (NF-κB) transcription factor to induce pyroptosis. Conclusions:The findings in this study reveal that potassium iodide promotesthe pyroptosis of thyroid follicular cells through activating NF-κB-NLRP3 inflammasome, which may be a novel mechanism that promots the development of Hashimoto′s thyroiditis under the condition of excessive iodine intake. PARP1 is a pivotal protein that mediates the pyroptosis induced by potassium iodide and may be a potential therapeutic target to control Hashimoto′s thyroiditis progression.
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【Objective】 To explore the role of RYBP in activating PARP-1 dependent Parthanatos and promoting response to YM155 in esophageal squamous cell carcinoma. 【Methods】 CCK-8 and flow cytometry were used to analyze the inhibition ratio and cell death percentage after YM155 treatment in both RYBP overexpression group and control group. Western blotting was used to detect the expression of Parthanatos-related proteins. 【Results】 Compared with control group, RYBP overexpression group showed higher inhibition ratio and cell death percentage after YM155 treatment. Overexpression of RYBP activated PARP-1 with or without YM155 treatment. Besides, after YM155 treatment, KYSE170-RYBP showed more PAR accumulation in the nucleus, AIF translocation from mitochondria to the nucleus than control cells. 【Conclusion】 RYBP can activate PARP-1/PAR/AIF-dependent induced Parthanatos in esophageal squamous cell carcinoma and enhance response to YM155.
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The chronic consumption of alcohol causes a worsening of the events that follow the cerebral ischemia. These events are regulated through the expression of several genes and microRNAs. The aimof this work was To analyze and describe the expression profile of PARP and AIF and miRNA-9 proteins in rats submitted to focal cerebral ischemia, associated or not with chronic alcoholism model. Methods: Twenty adult Wistar rats, subdivided into: control; ischemic; alcoholic and ischemic / alcoholized for immunohistochemical analysis and miRNA-9 gene expression. Results: There was a reduction in the protein expression of PARP-1 and a positive marking for AIF in the ischemic / alcoholized group. The miRNA-9 did not obtain significant expression. The association of ischemia with chronic alcohol use promoted a tendency to low expression of miRNA-9, low expression of PARP-1 and high expression of AIF, indicating an interference in the protective effect of miRNA-9 be observed in the other groups.
El consumo crónico de alcohol provoca un empeoramiento de los eventos que siguen a la isquemia cerebral. Estos eventos están regulados a través de la expresión de varios genes y microRNA. El objetivo de este trabajo fue analizar y describir el perfil de expresión de las proteínas PARP y AIF y microRNA-9 en ratas sometidas a isquemia cerebral focal, asociadas o no, con el modelo de alcoholismo crónico. Veinte ratas Wistar adultas se dividieron en: grupo control, isquémico alcohólico, e isquémico / alcoholizado para análisis inmunohistoquímico y expresión de genes microRNA-9. Resultados: Hubo una reducción en la expresión de proteínas de PARP-1 y un marcado positivo para AIF en el grupo isquémico / alcoholizado. No se observó una expresión significativa en el microRNA-9. La asociación de la isquemia con el consumo crónico de alcohol promovió una tendencia a la baja expresión de microRNA-9, baja expresión de PARP1 y alta expresión de AIF, lo que indica una interferencia en el efecto protector de microRNA-9 en los otros grupos.
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Animales , Ratas , Isquemia Encefálica/metabolismo , Alcoholismo/metabolismo , Inmunohistoquímica , Isquemia Encefálica/genética , Ratas Wistar , MicroARNs/metabolismo , Modelos Animales de Enfermedad , Alcoholismo/genética , Factor Inductor de la Apoptosis/metabolismo , Poli(ADP-Ribosa) Polimerasa-1/metabolismoRESUMEN
@#Poly(ADP-ribose)polymerase-1(PARP-1)plays a vital role in the regulation of DNA repair and apoptosis. Breakthrough has been made in the treatment of cancer with PARP-1 inhibitors, but the emergence of drug resistance has limited its further application in clinic. This paper reviews advances in the research on PARP-1 inhibitors combined with other drugs to overcome drug resistance, highlights and evaluates the existing drug combination strategies and their therapeutic effects in clinical practice. It is proposed that the development of dual-target or multi-target drugs will become a promising approach to overcome the resistance of PARP-1 inhibitors and broaden their indications.
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Aim To investigate the interaction and mechanism of PARP1 and NFATc3, NFATc4 in ISO-induced pathological cardiac hypertrophy. Methods To establish the model of cardiac hypertrophy in vitro and in vivo, primary neonatal rat cardiomyocytes were treated with ISO (10 jimol • L-1) for 24 h; SD rats were subcutaneously injected with 1. 2 mg • kg-1 • d-1 ISO for 7 d. The nuclear and cytoplasmic proteins were separated by Cellytic Nuclear Extraction Kit. The subcellular localization of NFATc3 and NAFTc4 were detected by Western blot and immunofluorescence. The recombinant adenovirus (Ad-PARPl) infection was used to overexpress PARP1 and knockdown PARP1 by transfecting with siRNA of PARP1 in cardiomyocytes. Results The models of cardiac hypertrophy were successfully built both in vivo and vitro by ISO. It was determined that NFATc3 and NFATc4 were transfered into the nuclear from the cytoplasm in primary neonatal rat cardiomyocytes (NRCMs) after being treated with ISO. And the enzymatic activity of PARP1 was boosted in TSO-trpatpH prmin. OvprpYnrp.ssinn of PARP1 nromo-ted the nuclear translocation of NFATc3 and NFATc4 in cardiomyocytes, while knockdown of PARP1 could reverse the nuclear translocation induced by ISO. PARP1 inhibitor 3AB retarded ISO-induced nuclear transportation of NFATc3 and NFATc4 to some extent. Conclusions ISO leads to the up-regulation of enzymatic activity of PARP1 and promotes nuclear translocation of NFATc3 and NFATc4, thus aggravating car-diac hypertrophy.
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OBJECTIVE To investigate the protective effect of olaparib on the inflammatory damage to alveolar epithelial cells induced by lipopolysaccharide (LPS). METHODS The alveolar epithelial cells (A549) were cultured in vitro and incubated with LPS 10 mg-L-1 and olaparib 10 and 25 μmol-L-1 for 24 h. The levels of cytokines interleukin 6 (IL-6), IL-8, and IL-10 were detected by enzyme linked immunosorbent assay (ELISA), the mRNA expression levels of TNF-α, IL-1β, IL-6, IL-8, and ICAM-1 were analyzed by real-time PCR, the level of ROS was analyzed by flow cytometry, and the expression of poly (ADP-ribose) polymerase-1 (PARP-1) and phosphorylation of proteins involved in NF-ΚB signaling pathway in cells were detected by Western blotting. RESULTS Compared with LPS 10 mg-L-1 injury group, olaparib 10 and 25 μmol·L-1 could significantly reduce the release of IL-6, IL-8 and ROS levels in A549 cells induced by LPS (P<0.01), and increase the release of IL-10 (P<0.01). Olaparib 10 and 25 μmol·L-1 could also inhibit the mRNA expressions of TNF-α, IL-1β, IL-6, IL-8, and ICAM-1 (P<0.01), and inhibit the expression of PARP-1 and phosphorylation proteins involved in NF-ΚB signaling pathway induced by LPS (P<0.01). CONCLUSION Olaparib has some protective effect on inflammatory damage and oxidative stress in alveolar epithelial cells induced by LPS, and the mechanism may be that it inhibits the expression and release of cytokines by down-regulating the expression of PARP-1 and subsequently affecting the activation of the NF-ΚB pathway.
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[Objective]To investigate whether IRF5 can inhibit invasion ability of nasopharyngeal carcinoma by re-ducing PARP-1(poly(ADP-ribose)polymerase-1).[Methods]Forty-six specimens of nasopharyngeal carcinoma and 51 specimens of normal tissue were confirmed by pathologically in this study.The expression of IRF5 and PARP-1 in naso-pharyngeal carcinoma tissues and normal tissues was detected by immunohistochemistry.The IFR5 overexpression plasmid was transfected into the nasopharyngeal carcinoma cell line CNE-2,quantitative PCR and immunoblotting was used to value the expression of IRF5 after transfection.The wound healing and transwell assay was used to investigate the invasion ability. The expression of PARP-1 was valued by quantitative PCR and immunoblotting after over-expression of PFR5.[Results]The results showed that the expression of IRF5 in cancer tissues was lower than that in normal tissues,but the PARP-1 expression was opposite. The IRF5 overexpressing cell line CNE-2/IFR5 was established. The healing rate of CNE-2/IFR5 cells was lower than that of the control cells(P<0.01). Transwell experiments revealed that the number of CNE-2/IFR5 cells passing through the basement membrane was smaller than that of the control group(P<0.01),suggest-ing that up-regulation of IFR5 could inhibit the invasiveness of nasopharyngeal carcinoma cells.Over-expression of IFR5 led to reduced PARP-1 mRNA and protein(P<0.01).Besides,elevation of PARP-1 can prevent IRF5-induced changes of invasion ability.[Conclusion]Therefore,we speculated that IRF5 can inhibit invasion ability of nasopharyngeal carci-noma by reducing the expression of PARP-1.This study provided a new target for inhibiting the invasion ability of naso-pharyngeal carcinoma based on IRF5.
RESUMEN
Poly(ADP-ribose)polymerase-1(PARP-1) is a ribozyme widely expressed in eukaryotic cells with significant biological activity, the activation of which is involved in the signal transduction of inflammatory pulmonary disease. In this paper, we review PARP-1 involvement in lung inflammation regulation and its prospects for treatment of diseases in terms of the structure and function of PARP-1, inflammation of the signal conditioning,regulation of lung inflammation disease and progress in inhibitor research. This review is intended to further clarify the pathogenesis of pulmonary inflammatory disease, and to provide reference for the prevention and treatment of the disease.