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SUMMARY OBJECTIVE: This study aimed to investigate the presence of indoleamine-2,3-dioxygenase and bacterial translocation after the administration of 3-aminobenzamide and infliximab in the TNBS model of rat colitis. METHODS: The study group was divided into five categories as follows: group 1: (control), group 2: colitis+saline, group 3: colitis+3-aminobenzamide, group 4: colitis+infliximab, and group 5: colitis+3-aminobenzamide+infliximab. Intestinal mesenteric cultures were incubated on specific agar media plates under aerobic and anaerobic conditions, bacterial translocation was evaluated and assessed as colony-forming units per gram of tissue. Colonic tissue samples were evaluated by Western blotting method to detect the presence of indoleamine-2,3-dioxygenase. RESULTS: The results obtained were as follows: group 1: normal gut flora; group 2: eight of nine samples had bacterial translocation, of which six of them had positive indoleamine-2,3-dioxygenase protein; group 3: five of nine samples had bacterial translocation, of which seven of them had positive indoleamine-2,3-dioxygenase; group 4: three of nine samples had bacterial translocation, of which seven of them had positive indoleamine-2,3-dioxygenase; and group 5: only one sample had exact indoleamine-2,3-dioxygenase protein. CONCLUSION: Altered expression of indoleamine-2,3-dioxygenase results in a lower bacterial translocation via infliximab compared with 3-aminobenzamide treatment. Combined treatments emphasized different approaches for the new molecules related to indoleamine-2,3-dioxygenase.
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The clinical application of doxorubicin (DOX) in cancer chemotherapy is limited by its life-threatening cardiotoxic effects. Chrysophanol (CHR), an anthraquinone compound isolated from the rhizome of L., is considered to play a broad role in a variety of biological processes. However, the effects of CHR׳s cardioprotection in DOX-induced cardiomyopathy is poorly understood. In this study, we found that the cardiac apoptosis, mitochondrial injury and cellular PARylation levels were significantly increased in H9C2 cells treated by Dox, while these effects were suppressed by CHR. Similar results were observed when PARP1 activity was suppressed by its inhibitors 3-aminobenzamide (3AB) and ABT888. Ectopic expression of PARP1 effectively blocked this CHR׳s cardioprotection against DOX-induced cardiomyocyte injury in H9C2 cells. Furthermore, pre-administration with both CHR and 3AB relieved DOX-induced cardiac apoptosis, mitochondrial impairment and heart dysfunction in Sprague-Dawley rat model. These results revealed that CHR protects against DOX-induced cardiotoxicity by suppressing cellular PARylation and provided critical evidence that PARylation may be a novel target for DOX-induced cardiomyopathy.
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Background & objectives: Drug resistance is the primary cause of failure in the treatment of cancers. It has been suggested that the enhancement of DNA repair capability may be responsible for the drug resistance of the tumour cells, and poly(ADP-ribosyl)ation plays an important role in DNA repair. This study investigated the effect of PARP inhibitor 3-aminobenzamide (3-AB) on the cisplatin resistance and proliferation of the cisplatin-resistant ovarian cancer C13* cells in vitro. Methods: C13* cells were treated with various concentrations of 3-AB in vitro. MTT assay was used to determine the effect of 3-AB on the cisplatin sensitivity and proliferation of cells. The expression levels of PARP-1 mRNA and protein in the C13* cells were examined using reverse transcription-polymerase chain reaction (RT-PCR) and Western blot, and changes caused by 3-AB treatment were investigated. Immunofluorescence microscopy was used to detect the localization and expression of the PARP-1 proteins before and after treatment with 5 mmol/l 3-AB. Results: The inhibitory ratio and the cisplatin sensitivity of C13* cells significantly increased with the increase of the concentration of 3-AB (P<0.05). The RT-PCR analysis revealed that the expression of PARP-1 mRNA was decreased when platinum (Pt) and 3-AB were combined. The expression levels of PARP-1 protein were decreased by 23.15 ± 2.53, 59.11 ± 2.23 and 73.24 ± 3.88 per cent, respectively, in C13* cells with the increase of the concentration of 3-AB (P<0.05). The immunofluorescence microscopy results indicated that the expression level of PARP-1 protein was significantly decreased after treatment with 3-AB (P<0.05). Interpretation & conclusions: 3-AB inhibited the proliferation activity of C13* cells, and increased the cellular sensitivity to cisplatin. Our findings show that the PARP inhibitor 3-AB can downregulate the expression of PARP-1 at transcriptional and translational levels in C13* cells.
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Objective To evaluate the protection of 3-aminobenzamide (3-AB),an inhibitor of Poly (ADP-ribose) polymerase (PARP),on severe acute pancreatitis associated adenohypophysis injury in rats.Method Forty Wistar rats were randomly divided into 4 groups:sham operation group (SO group,n=8),SAP group (n=12),3-AB pretreatment group (n =12),drug control group (n =8).The bilepancreatic duct was cannulated through the duodenum and SAP model was induced by a standardized pressure-controlled retrograde infusion of 5％ sodium taurocholate (0.1 ml/100 g) into the bile-pancreatic duct.In 3-AB group,3-AB (20 mg/kg) was administered via femoral vein 30 min prior to the operation;other procedures were identical to SAP group.In SO group,pancreas was flipped several times only.In drug control group,3-AB (20 mg/kg) was administered via femoral vein 30 min prior to the operation.Serum amylase,lipase were measured.Pancreas and pituitary tissue were taken for pathological examination under light microscope.PARP and NF-κB antibodies for adenohypophysis immunohistochemical stains.Adenohypophysis cell was observed under electronic microscope.Result Serum amylase,lipase and pancreas pathological scores were significantly higher in 3-AB group compared with SO group (P ＜ 0.05),but lower than that in SAP group (P ＜ 0.05).Adenohypophysis pathological injury was less severe in 3-AB group.Expressions of PARP and NF-κB in adenohypophysis cells were significantly higher in 3-AB group compared with SO group,but lower than that in SAP group (P ＜ 0.05).Ultrastructural change of thyrotroph cell was relieved in 3-AB group.No significant difference was observed between SO group and drug control group in PARP and NF-κB expression nor adenohypophysis pathological injury.Conclusions 3-AB exerts the protective effect against acute pancreatitis associated adenohypophysis injury by inhibition of PARP and NF-κB.
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Objective To investigate the effects and mechanisms of poly adenosine diphosphate (ADP)-ribose polymerase inhibitor 3-aminobenzamide (3-AB) on kidney injury in rates with severe acute pancreatitis (SAP).Methods Fifty-six male Wistar rats were divided into the sham operation (SO) group,SAP (3,6,12 hours) groups,and 3-AB + SAP (3,6,12 hours) groups,and there were 8 rats in each group.SAP model was established by retrograde injection of 5％ sodium taurocholate into the biliopancreatic duct.Rats in the 3-AB + SAP group were infused with 3-AB (20 μg/g) via femoral vein 30 minutes before SAP model establishment.The serum amylase,kidney function and renal myeloperoxidase (MPO) were determined,and pathological scores of pancreatic and renal tissues were evaluated under light microscope.Renal poly ADP-ribose formation,intercellular adhesion molecules-1 (ICAM-1) and P-selectin expression were detected by the Western blot.All data were analyzed using the analysis of variance or t test.Renal injury grading was analyzed using the Kruskal-Wallis nonparametric test.Results The levels of serum amylase of SAP 3,6,12 groups were (3806 ± 229)U/L,(4898 ± 295) U/L and (5726 ± 372) U/L,which were significantly higher than (2785 ± 160) U/L,(3241 ± 198) U/L and (3953 ± 249) U/L of the 3-AB + SAP groups (t =3.652,4.672,4.407,P ＜ 0.05).The levels of blood urea nitrogen were (11.6 ± 0.8) mmol/L,(19.3 ± 1.3) mmol/L and (29.6 ± 2.1) mmol/L,which were higher than (7.5 ± 0.5) mmol/L,(10.5 ± 0.7) mmol/L and (21.6 ± 1.5) mmol/L of the 3-AB + SAP groups.There were significant differences in the levels of blood urea nitrogen between the SAP group and the 3-AB + SAP group at the 6 and 12 hours (t =3.836,6.849,P ＜0.05).The levels of creatinine of the SAP 3,6,12 hours groups were (48.7 ±3.1) μmol/L,(58.3 ±3.7) μmol/L and (75.9 ±5.4) μmol/L,which were higher than (40.7 ±2.6)μmol/L,(43.2 ± 2.6) μmol/L and (53.4 ± 3.2) μmol/L of the 3-AB + SAP groups.There were significant differences in the levels of creatinine between the SAP group and the 3-AB + SAP group at the 6 and 12 hours (t =3.279,3.073,P ＜ 0.05).The renal MPO activity of the SAP 3,6,12 hours groups were (0.69 ± 0.06) U/g,(1.07 ± 0.09)U/g and (1.42 ±0.13)U/g,which were higher than (0.57 ±0.05)U/g,(0.75 ±0.06)U/g and (0.89 ± 0.07) U/g of the 3-AB + SAP groups.There were significant differences in the renal MPO activity between the SAP group and the 3-AB + SAP group at the 6 and 12 hours (t =3.066,4.012,P ＜ 0.05).The pancreatic pathological scores of the SAP 3,6 and 12 hours group were 6.50 ± 0.53,9.06 ± 0.66 and 11.75 ± 0.89,which were significantly higher than 4.25 ± 0.31,6.06 ± 0.51 and 7.57 ± 0.59 of the 3-AB + SAP group (t =3.631,3.598,5.147,P ＜ 0.05).The structure of the kidney was normal in the SO group.Congestive changes were observed in glomerulus of kidney,the renal tubular epithelial cell was necrosed,and luminal narrowing or occlusion,hemorrhage in the intercellular substance and inflammatory cell infiltration were observed in the SAP 12 hours group.The pathological changes of the 3-AB + SAP 12 hours group were significantly slighter (P ＜ 0.05).The relative expressions of poly ADP-ribose,ICAM-1 and P-selectin of the SO group were 1.00 ±0.21,1.00 ±0.18,1.00 ± 0.16,which were significantly lower than 3.83 ± 0.63,5.42 ± 0.83,3.71 ± 0.48 of the SAP 12 hours group (t =6.955,23.107,10.352,P ＜ 0.05).The relative expressions of poly-ADP-ribose,ICAM-1 and P-selectin of the 3-AB + SAP 12 hours group were 1.94 ± 0.36,2.35 ± 0.35,2.11 ± 0.29,which were significantly lower than SAP 12 hours group (t =3.977,12.115,5.012,P ＜ 0.05).Conclusions Poly ADP-ribose polymerase inhibitor 3-AB protects kidney from injury in the experimental SAP rats.Poly ADP-ribose polymerase inhibitor 3-AB functions by suppressing the ICAM-1 and P-selectin expression and reducing neutrophil infiltration.
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Objective To investigate the protective effects of poly-ADP-ribose polymerase inhibitor 3-aminobenzamide (3-AB) on lung injury in rats with severe acute pancreatitis (SAP) and to explore the mechanisms.Methods Thirty-six Wistar rats were randomly (random number) divided into three groups (n =12 for each group),namely sham operation (SO) group,SAP group and 3-AB-treated group.The model of SAP-associated lung injury was established by retrograde injection of 5％ sodium taurocholate (STC) into the biliopancreatic duct.In the treated group,3-AB in dose of 10 mg/kg was administered twice by intravenous injection 30 min before and 30 min after STC infusion.The survival rats were sacrificed 12hours after SAP modeling,and the serum amylase,lung wet/dry ratio and myeloperoxidase (MPO) activity were determined,and pathological scores of pancreas and lung tissue were evaluated under light microscope.Expressions of interleukin (IL) -1 β and IL-6 mRNA,tumor necrosis factor α (TNF-α) and inter-cellular adhesion molecule-1 (ICAM-1) protein were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot,respectively. Results The serum amylase level,lung wet/dry ratio and MPO activity,IL-1β and IL-6 mRNA expressions,TNF-α and ICAM-1 protein levels were dramatically increased in SAP group ( P ＜ 0.05 ).Treatment with 3-AB significantly reduced these biomarkers in 3-AB group than in SAP group (P ＜ 0.05 ).Conclusions Poly-ADP-ribose polymerase inhibitor 3-AB exerts the protective and therapeutic effects on lung injury associated with severe acute pancreatitis through inhibiting intrapulmonary MPO activity and down-regulating the expressions of IL-1 β and IL-6 mRNA as well as the levels of TNF-α,and ICAM-1.
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Objective To evaluate the ability of 3-AB to sensitize the human esophageal carcinoma cell strain (CaEs-17) to radiation in v/tro and its mechanisms. Methods CaEs-17 cells were treated with 3-AB at 0, 2.5, 7.5 mmol/L and given irradiation O, 3, 6, 9, 12 Gy. 3-AB concentration in each group was made dose-survival curve using multi-target single-hit maiths model by clonogenie assay. MTT assay was performed to observe the survival of irradiated cells.comet assay and metaphase chromosome analysis were used to measure the DNA damage degree and chromosome aberration of CaEs-17 cell after 3-AB treatment and irradiation. Results Cell survival experiments showed SER of 1.21, 1.52 for 2.5 mmol/L, 7.5 mmol/L 3-AB respectively using multi-target single-hit maths model. The survival fraction of irradiated CaEs-17 cell was decreased after 3-AB treatment. DNA damage and the chromatid breakage number of irradiated CaEs-17 cells were increased after 3-AB treatment. Conclusions 3-AB, a PARP inhibitor, can enhance the radiosensitivity of human esophageal carcinoma cell strain (CaEs-17). DNA damage repair inhibition by 3-AB might be one of the mechanisms.
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3-Nitropropionic acid (3-NP) inhibits electron transport in mitochondria, leading to a metabolic failure. In order to elucidate the mechanism underlying this toxicity, we examined a few biochemical changes possibly involved in the process, such as metabolic inhibition, generation of reactive oxygen species (ROS), DNA strand breakage, and activation of Poly (ADP-ribose) polymerase (PARP). Exposure of SK-N-BE (2) C neuroblastoma cells to 3-NP for 48 h caused actual cell death, while inhibition of mitochondrial function was readily observed when exposed for 24 h to low concentrations (0.2~2 mM) of 3-NP. The earliest biochemical change detected with low concentration of 3-NP was an accumulation of ROS (4 h after 3-NP exposure) followed by degradation of DNA. PARP activation by damaged DNA was also detectable, but at a later time. The accumulation of ROS and DNA strand breakage were suppressed by the addition of glutathione or N-acetyl-L-cysteine (NAC), which also partially restored mitochondrial function and cell viability. In addition, inhibition of PARP also reduced the 3-NP-induced DNA strand breakage and cytotoxicity. These results suggest that oxidative stress and activation of PARP are the major factors in 3-NP-induced cytotoxicity, and that the inhibition of these factors may be useful in protecting neuroblastoma cells from 3-NP-induced toxicity.