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1.
Neuropharmacology ; 245: 109818, 2024 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-38142931

RESUMO

Cardiac autonomic neuropathy resulting from human immunodeficiency virus (HIV) infection is common; however, its mechanism remains unknown. The current work attempted to explore the function and mechanism of the P2Y13 receptor in HIV-glycoprotein 120 (gp120)-induced neuropathy in cervical sympathetic ganglion. The superior cervical ganglion (SCG) of the male SD rat was coated with HIV-gp120 to establish a model of autonomic neuropathy. In each group, we measured heart rate, blood pressure, heart rate variability, sympathetic nerve discharge and cardiac function. The expression of P2Y13 mRNA and protein in the SCG was tested by real-time polymerase chain reaction and western blotting. Additionally, this study focused on identifying the protein levels of NOD-like receptor family pyrin domain-containing 3 (NLRP3), Caspase-1, Gasdermin D (GSDMD), interleukin (IL)-1ß and IL-18 in the SCG using western blotting and immunofluorescence. In gp120 rats, increased blood pressure, heart rate, cardiac sympathetic nerve activity, P2Y13 receptor levels and decreased cardiac function could be found. P2Y13 shRNA or MRS2211 inhibited the above mentioned changes induced by gp120, suggesting that the P2Y13 receptor may be engaged in gp120-induced sympathetic nerve injury. Moreover, the levels of NLRP3, Caspase-1, GSDMD, IL-1ß and IL-18 in the gp120 group were increased, while significantly decreased by P2Y13 shRNA or MRS2211. Therefore, the P2Y13 receptor is involved in gp120-induced sympathetic neuropathy, and its molecular mechanism shows an association with the activation of the NLRP3 inflammasome, followed by GSDMD formation along with the release of inflammatory factors including IL-1ß and IL-18. This article is part of the Special Issue on "Purinergic Signaling: 50 years".


Assuntos
Infecções por HIV , HIV-1 , Doenças do Sistema Nervoso Periférico , Receptores Purinérgicos P2 , Animais , Masculino , Ratos , Proteínas de Transporte , Caspases , Glicoproteínas/metabolismo , Infecções por HIV/complicações , Infecções por HIV/metabolismo , Inflamassomos/metabolismo , Interleucina-18/metabolismo , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Doenças do Sistema Nervoso Periférico/virologia , Ratos Sprague-Dawley , RNA Interferente Pequeno , Gânglio Cervical Superior/metabolismo , Proteína gp120 do Envelope de HIV/metabolismo , Receptores Purinérgicos P2/metabolismo
2.
Front Oncol ; 13: 1153319, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37182180

RESUMO

Non-small cell lung cancer (NSCLC) is a prominent etiology of cancer-related mortality. The heterogeneous nature of this disease impedes its accurate diagnosis and efficacious treatment. Consequently, constant advancements in research are imperative in order to comprehend its intricate nature. In addition to currently available therapies, the utilization of nanotechnology presents an opportunity to enhance the clinical outcomes of NSCLC patients. Notably, the burgeoning knowledge of the interaction between the immune system and cancer itself paves the way for developing novel, emerging immunotherapies for treating NSCLC in the early stages of the disease. It is believed that with the novel engineering avenues of nanomedicine, there is a possibility to overcome the inherent limitations derived from conventional and emerging treatments, such as off-site drug cytotoxicity, drug resistance, and administration methods. Combining nanotechnology with the convergence points of current therapies could open up new avenues for meeting the unmet needs of NSCLC treatment.

3.
Neurochem Res ; 48(3): 781-790, 2023 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-36331667

RESUMO

Neuropathic pain (NP) is a type of chronic pain affecting 6-8% of human health as no effective drug exists. The purinergic 2X4 receptor (P2X4R) is involved in NP. Neohesperidin (NH) is a dihydroflavonoside compound, which has anti-inflammatory and antioxidative properties. This study aimed to investigate whether NH has an effect on P2X4R-mediated NP induced by chronic constriction injury (CCI) of the sciatic nerve in rats. In this study, the CCI rat model was established to observe the changes of pain behaviors, P2X4R, and satellite glial cells (SGCs) activation in dorsal root ganglion (DRG) after NH treatment by using RT-PCR, immunofluorescence double labeling and Western blotting. Our results showed CCI rats had mechanical and thermal hyperalgesia with an increased level of P2X4R. Furthermore, SGCs were activated as indicated by increased expression of glial fibrillary acidic protein and increased tumor necrosis factor-alpha receptor 1and interleukin-1ß. In addition, phosphorylated extracellular regulated protein kinases and interferon regulatory factor 5 in CCI rats increased. After NH treatment in CCI rats, the levels of above protein decreased, and the pain reduced. Overall, NH can markedly alleviate NP by reducing P2X4R expression and SGCs activation in DRG.


Assuntos
Neuralgia , Receptores Purinérgicos P2X4 , Ratos , Humanos , Animais , Ratos Sprague-Dawley , Receptores Purinérgicos P2X4/metabolismo , Neuroglia/metabolismo , Hiperalgesia/metabolismo , Neuralgia/metabolismo , Gânglios Espinais/metabolismo
4.
Bioorg Chem ; 127: 105928, 2022 10.
Artigo em Inglês | MEDLINE | ID: mdl-35717802

RESUMO

The superbug infection mediated by metallo-ß-lactamases (MßLs) has grown into anemergent health threat, and development of MßL inhibitors is an ideal strategy to combat the infection. In this work, twenty-five thiosemicarbazones 1a-e, 2a-e, 3a-e, 4a-d, 5a-d and 6a-b were synthesized and assayed against MßLs ImiS, NDM-1 and L1. The gained molecules specifically inhibited NDM-1 and ImiS, exhibiting an IC50 value in the range of 0.37-21.35 and 0.45-8.76 µM, and 2a was found to be the best inhibitor, with an IC50 of 0.37 and 0.45 µM, respectively, using meropenem (MER) as substrate. Enzyme kinetics and dialysis tests revealed and confirmed by ITC that 2a is a time-and dose-dependent inhibitor of ImiS and NDM-1, it competitively and reversibly inhibited ImiS with a Ki value of 0.29 µM, but irreversibly inhibited NDM-1. Structure-activity relationship disclosed that the substitute dihydroxylbenzene significantly enhanced inhibitory activity of thiosemicarbazones on ImiS and NDM-1. Most importantly, 1a-e, 2a-e and 3a-b alone more strongly sterilized E. coli-ImiS and E. coli-NDM-1 than the MER, displaying a MIC value in the range of 8-128 µg/mL, and 2a was found to be the best reagent with a MIC of 8 and 32 µg/mL. Also, 2a alone strongly sterilized the clinical isolates EC01, EC06-EC08, EC24 and K. pneumonia-KPC-NDM, showing a MIC value in the range of 16-128 µg/mL, and exhibited synergistic inhibition with MER on these bacteria tested, resulting in 8-32-fold reduction in MIC of MER. SEM images shown that the bacteria E. coli-ImiS, E. coli-NDM-1, EC24, K. pneumonia-KPC and K. pneumonia-KPC-NDM treated with 2a (64 µg/mL) suffered from distortion, emerging adhesion between individual cells and crumpled membranes. Mice tests shown that monotherapy of 2a evidently limited growth of EC24 cells, and in combination with MER, it significantly reduced the bacterial load in liver and spleen. Docking studies suggest that the 2,4-dihydroxylbenzene of 2a acts as zinc-binding group with the Zn(II) and the residual amino acids in CphA active center, tightly anchoring the inhibitor at active site. This work offered a promising scaffold for the development of MßLs inhibitors, specifically the antimicrobial for clinically drug-resistant isolates.


Assuntos
Tiossemicarbazonas , Inibidores de beta-Lactamases , Animais , Antibacterianos/química , Antibacterianos/farmacologia , Bactérias/metabolismo , Escherichia coli , Camundongos , Testes de Sensibilidade Microbiana , Tiossemicarbazonas/farmacologia , Inibidores de beta-Lactamases/química , Inibidores de beta-Lactamases/farmacologia , beta-Lactamases/metabolismo
5.
Bioorg Chem ; 124: 105799, 2022 07.
Artigo em Inglês | MEDLINE | ID: mdl-35462235

RESUMO

The emerging COVID-19 pandemic generated by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) has severely threatened human health. The main protease (Mpro) of SARS-CoV-2 is promising target for antiviral drugs, which plays a vital role for viral duplication. Development of the inhibitor against Mpro is an ideal strategy to combat COVID-19. In this work, twenty-three hydroxamates 1a-i and thiosemicarbazones 2a-n were identified by FRET screening to be the potent inhibitors of Mpro, which exhibited more than 94% (except 1c) and more than 69% inhibition, and an IC50 value in the range of 0.12-31.51 and 2.43-34.22 µM, respectively. 1a and 2b were found to be the most effective inhibitors in the hydroxamates and thiosemicarbazones, with an IC50 of 0.12 and 2.43 µM, respectively. Enzyme kinetics, jump dilution and thermal shift assays revealed that 2b is a competitive inhibitor of Mpro, while 1a is a time-dependently inhibitor; 2b reversibly but 1a irreversibly bound to the target; the binding of 2b increased but 1a decreased stability of the target, and DTT assays indicate that 1a is the promiscuous cysteine protease inhibitor. Cytotoxicity assays showed that 1a has low, but 2b has certain cytotoxicity on the mouse fibroblast cells (L929). Docking studies revealed that the benzyloxycarbonyl carbon of 1a formed thioester with Cys145, while the phenolic hydroxyl oxygen of 2b formed H-bonds with Cys145 and Asn142. This work provided two promising scaffolds for the development of Mpro inhibitors to combat COVID-19.


Assuntos
Tratamento Farmacológico da COVID-19 , Tiossemicarbazonas , Animais , Antivirais/química , Proteases 3C de Coronavírus , Humanos , Camundongos , Simulação de Acoplamento Molecular , Pandemias , Inibidores de Proteases/química , SARS-CoV-2 , Tiossemicarbazonas/farmacologia
6.
Bioorg Chem ; 120: 105654, 2022 03.
Artigo em Inglês | MEDLINE | ID: mdl-35149265

RESUMO

The "superbug" infection caused by metallo-ß-lactamases (MßLs) has grown into anemergent health threat, and development of effective MßL inhibitors to restore existing antibiotic efficacy is an ideal alternative. Although the serine-ß-lactamase inhibitors have been used in clinical settings, MßL inhibitors are not available to date. In this work, thirty-one quinolinyl sulfonamides 1a-p and sulphonyl esters 2a-o were synthesized and assayed against MßL NDM-1. The obtained molecules specifically inhibited NDM-1, 1a-p and 2a-o exhibited an IC50 value in the range of 0.02-1.4 and 8.3-24.8 µM, respectively, and 1e and 1f were found to be the most potent inhibitors, with an IC50 of 0.02 µM using meropenem (MER) as substrate. Structure-activity relationship reveals that the substitute phenyl and the phenyl with a halogen atom more significantly improve inhibitory effect of quinolinederivatives on NDM-1. 1a-p restored antimicrobial effect of MER on E. coli with NDM-1, EC01 and EC08, resulting in a 2-64-fold reduction in MIC values. Most importantly, 1e synergized MER and significantly reduced the load of EC08 in the spleen and liver of mice after a single intraperitoneal dose. Docking studies suggested that the endocyclic nitrogen of the quinoline ring, and exocyclic nitrogen of the sulfonamide functional group are coordinate with Zn(II) ion at active sites of NDM-1. Cytotoxicity assays indicated that 1e had low cytotoxicity. This work offers potential lead compounds for further development of the clinically useful inhibitor targeting NDM-1.


Assuntos
Escherichia coli , Ésteres , Animais , Antibacterianos/química , Antibacterianos/farmacologia , Ésteres/farmacologia , Camundongos , Testes de Sensibilidade Microbiana , Nitrogênio/farmacologia , Sulfanilamida/farmacologia , Sulfonamidas/farmacologia , beta-Lactamases/química
7.
Chem Biol Drug Des ; 99(2): 362-372, 2022 02.
Artigo em Inglês | MEDLINE | ID: mdl-34862744

RESUMO

Bacterial resistance caused by metallo-ß-lactamases (MßLs) has become an emerging public health threat, and the development of MßLs inhibitor is an effective way to overcome the resistance. In this study, thirteen novel O-aryloxycarbonyl hydroxamates were constructed and assayed against MßLs. The obtained molecules specifically inhibited imipenemase-1 (IMP-1) and New Delhi metallo-ß-lactamase-1, exhibiting an IC50 value in the range of 0.10-18.42 and 0.23-22.33 µM, respectively. The hydroxamate 5 was found to be the most potent inhibitor, with an IC50 of 0.1 and 0.23 µM using meropenem and cefazolin as substrates. ICP-MS analysis showed that 5 did not coordinate to the Zn(II) ions at the active site of IMP-1, while the rapid dilution, thermal shift and MALDI-TOF assays revealed that the hydroxamate formed a covalent bond with the enzyme. Cytotoxicity assays indicated that the hydroxamates have low toxicity in MCF-7 cells. This work provided a potent scaffold for the development of MßLs inhibitors.


Assuntos
Ácidos Hidroxâmicos/química , Inibidores de beta-Lactamases/química , Espectroscopia de Ressonância Magnética Nuclear de Carbono-13 , Humanos , Células MCF-7 , Espectroscopia de Prótons por Ressonância Magnética , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Relação Estrutura-Atividade , Inibidores de beta-Lactamases/farmacologia
8.
Mol Med Rep ; 24(4)2021 10.
Artigo em Inglês | MEDLINE | ID: mdl-34396429

RESUMO

Following the publication of the above article, the authors have realized that, on p. 8, some of the supplementary data were cited incorrectly in the main text. In the right-hand column, second paragraph, the sentence beginning on line 5 should have read as follows (changed text is highlighted in bold): "Meanwhile, the expression of miR­513b­5p in tumor tissues was decreased and the expression of PRPF39 was increased in tumor tissues with knockout of circ­G004213 (Fig. S3D and E)." (i.e., the reference to Fig. S3C and D was incorrect.). The authors regret their oversight in failing to correct the inaccurate citation of the data in the paper, are grateful to the Editor for allowing them the opportunity to publish this Corrigendum, and apologize to the readership for any inconvenience caused. [the original article was published in Molecular Medicine Reports 23: Article no. 421, 2021; DOI: 10.3892/mmr.2018.12060].

9.
Bioorg Chem ; 114: 105138, 2021 09.
Artigo em Inglês | MEDLINE | ID: mdl-34229201

RESUMO

The expression of ß-lactamases, especially metallo-ß-lactamases (MßLs) in bacteria is one of the main causes of drug resistance. In this work, an effective N-acylhydrazone scaffold as MßL inhibitor was constructed and characterized. The biological activity assays indicated that the synthesized N-acylhydrazones 1-11 preferentially inhibited MßL NDM-1, and 1 was found to be the most effective inhibitor with an IC50 of 1.2 µM. Analysis of IC50 data revealed a structure-activity relationship, which is that the pyridine and hydroxylbenzene substituents at 2-position improved inhibition of the compounds on NDM-1. ITC and enzyme kinetics assays suggested that it reversibly and competitively inhibited NDM-1 (Ki = 0.29 ± 0.05 µM). The synthesized N-acylhydrazones showed synergistic antibacterial activities with meropenem, reduced 4-16-fold MIC of meropenem on NDM-1- producing E. coli BL21 (DE3), while 1 restored 4-fold activity of meropenem on K. pneumonia expressing NDM-1 (NDM-K. pneumoniae). The mice experiments suggested that 1 combined meropenem to fight against NDM-K. pneumoniae infection in the spleen and liver. Cytotoxicity assays showed that 1 and 2 have low cytotoxicity. This study offered a new framework for the development of NDM-1 inhibitors.


Assuntos
Hidrazonas/farmacologia , Inibidores de beta-Lactamases/farmacologia , beta-Lactamases/metabolismo , Animais , Chlorocebus aethiops , Relação Dose-Resposta a Droga , Hidrazonas/síntese química , Hidrazonas/química , Camundongos , Camundongos Endogâmicos , Estrutura Molecular , Relação Estrutura-Atividade , Células Vero , Inibidores de beta-Lactamases/síntese química , Inibidores de beta-Lactamases/química
10.
World J Clin Cases ; 8(8): 1554-1560, 2020 Apr 26.
Artigo em Inglês | MEDLINE | ID: mdl-32368550

RESUMO

BACKGROUND: Coronavirus disease 2019 (COVID-19) is a major public health emergency with obvious characteristics of human-to-human transmission, and there are infective asymptomatic carriers. Early identification and proper management of patients with COVID-19 are important. Features in chest computed tomography (CT) can facilitate identifying newly infected individuals. However, CT findings of some lung contusions are similar to those of COVID-19, as shown in the present case. CASE SUMMARY: A 46-year-old woman was admitted to hospital for backache and foot pain caused by a fall injury 1 d before hospitalization. She was suspected of having COVID-19, since there was a confirmed COVID-19 case near her residence. But she had no fever, cough, chest tightness, difficult breathing, nausea, vomiting, or diarrhea, etc. On physical examination, the lower posterior chest of both sides showed dullness on percussion and moist rales at the end of inspiration on auscultation. The white blood cell count and lymphocyte count were 10.88 × 109/L and 1.04 × 109/L, respectively. CT performed on February 7, 2020 revealed that both lungs were scattered with patchy ground-glass opacity. The patient was diagnosed with pulmonary contusion with thoracic spinal fracture (T12), calcaneal fracture, and pelvic fracture. On day 9 after conservative treatment, her condition was alleviated. On review of the chest CT, the previous shadows were significantly reduced. CONCLUSION: Differential diagnosis of lung contusion and COVID-19 must be emphasized. Both conditions require effective prompt actions, especially COVID-19.

11.
J Thorac Dis ; 11(8): 3421-3426, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31559046

RESUMO

BACKGROUND: The key for conducting thymectomy by thoracoscope is the treatment of thymic veins, as they are the most common source of bleeding. The traditional treatment is to cut off the distal and proximal vessels after Ham-Lock clamping. Our team found that it is safe and reliable to use ultrasonic energy to cut off thymic veins in previous open operations. This study aimed to investigate the feasibility and safety of thymic vein sealing with ultrasonic energy in video-assisted thoracoscopic surgery (VATS) thymectomy. METHODS: A total of 169 patients with or without thymic tumors who underwent thymectomy by thoracoscope were enrolled in the study. Among them, group A, with 89 patients, underwent thoracoscopic thymic vein resection by Ham-Lock, and group B, with 80 patients, did so by ultrasonic energy. The groups were compared in terms of the characteristics of patients, tumors, and perioperative period. RESULTS: There were no significant differences in patients' characteristics between the two groups (P>0.05). There was no significant difference between the two groups in blood transfusion, operation time, drainage, and hospitalization (P>0.05). There were no deaths, secondary operation and post-operative blood transfusion, and no serious complications leading to prolonged hospitalization. One patient in group A was converted to thoracotomy due to left anonymous venous hemorrhage, and one patient in group B had thymic venous hemorrhage, but the hemorrhage was successfully stopped under thoracoscope. CONCLUSIONS: In VATS thymectomy (with or without thymic tumors), there is no significant difference in the treatment of thymic veins between Ham-Lock clipping and ultrasonic energy in conversion to open surgery in regards to thymic venous hemorrhage, postoperative drainage, blood transfusion, operation time, drainage, and hospital stay. Use of ultrasonic energy is a safe and reliable method for thymic vein disconnection.

12.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-713041

RESUMO

@#Objective To analyze the feasibility and advantages of non-intubated anesthesia in thoracoscopic lobectomy. Methods The clinical data of 59 patients with thoracoscopic lobectomy and non-intubated anesthesia in the Department of Thoracic Surgery, Tongji Hospital from January 2015 to December 2017 were retrospectively reviewed, including 24 males and 35 females, aged 56.86±7.13 years (an observation group); 59 patients with thoracoscopic lobectomy undergoing general anesthesia with tracheal intubation in the same period were randomly selected, as a control group, including 27 males and 32 females, aged 55.37±6.86 years. Complications such as airway injury, refractory cough, pharyngalgia, nausea and vomiting were compared between the two groups. Postoperative inflammatory factor levels, postoperative hospital stay, and intraoperative and postoperative hospitalization costs were also compared. Results There was no difference between the two groups in general conditions such as age, gender, body mass index. There was also no difference in operation time, intraoperative bleeding volume or lymph node dissection. But the observation group had lower levels of procalcitonin and C reactive protein at postoperative 1 d (0.12±0.51 ng/ml vs. 0.14±0.70 ng/ml, P=0.03; 11.30±3.60 mg/L vs. 13.33±4.41 mg/L, P=0.01), lower rate of postoperative complications of refractory cough, pharyngalgia, nausea and vomiting (3.38% vs. 15.25%, P=0.03; 5.08% vs. 20.33%, P=0.01; 3.38% vs. 15.25%, P=0.03), less retain time of thoracic duct, postoperative hospital stay, and lower intraoperative and postoperative hospitalization costs (5.89±1.37 d vs. 7.00±1.73 d, P=0.00; 10.01±1.85 d vs. 11.37±2.45 d, P=0.00; 53 810.94±5 745.44 yuan vs. 58 223.16±6 445.08 yuan, P=0.00). Conclusion Thoracoscopic lobectomy with non-intubated anesthesia can avoid traditional airway injury caused by endotracheal intubation, reduce postoperative symptoms such as refractory cough, pharyngalgia, nausea and vomiting caused by general anesthesia, reduce or even avoid lung injury caused by one-side lung ventilation, promote recovery after surgery, reduce antibiotic use, and shorten hospital stay, which is more consistent with the requirements of the concept of overall minimal invasiveness and enhanced recovery.

13.
Mol Med Rep ; 18(6): 5302, 2018 12.
Artigo em Inglês | MEDLINE | ID: mdl-30320403

RESUMO

Following the publication of the above article, the authors have realized that the Materials and methods section and Figs. 1, 3 and 4 contained certain errors. In the Materials and methods section, the high glucose concentration was reported incorrectly as being 25 mmol/l; this should have been stated as 15 mmol/l. Furthermore, the normal concentration of glucose should have been reported as 5 mmol/l rather than  5.5 mmol, and the glucose concentration gradient should have been written as 0, 5, 10, and 15 mmol/l, and not as 0, 5, 10, and 25 mmol/l. Secondly, the authors have corrected some miscalculations associated with the bar charts featured in Figs. 1, 3, and 4, and the revised versions of these figures are shown here. All these corrections were approved by all authors. The authors regret that these errors were included in the paper, even though they did not substantially alter any of the major conclusions reported in the study, and apologize to the readership for any inconvenience caused. [the original article was published in Molecular Medicine Reports 17: 8536­8541, 2018; DOI: 10.3892/mmr.2018.8914].

14.
Mol Med Rep ; 17(6): 8536-8541, 2018 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-29693146

RESUMO

Over the past few decades, it has been demonstrated that hyperglycemia can promote lung carcinoma growth, potentially through significantly increased glucose metabolism; however, the underlying mechanism remains to be fully elucidated. In the present study, treatment with a high concentration of glucose (HG) significantly promoted the proliferation and migration of A549 cells. Receptor for advanced glycation end­products (RAGE) has previously been demonstrated to be associated with diabetes mellitus and oxidative stress, and nicotinamide adenine dinucleotide phosphate oxidases (NOXs) are considered to be initiating factors of oxidative stress. Therefore, an MTT assay, wound­healing assay, quantitative polymerase chain reaction and western blotting assays were used to analyze the RAGE­NOX­4 pathway and to determine its potential involvement in glycometabolism­associated tumorigenesis. The present study demonstrated that HG could increase the protein expression of RAGE and NOX­4, whereas the inhibitor of RAGE (anti­RAGE antibody) could suppress this effect. Futhermore, the inhibitor of NOX [diphenyl iodonium chloride (DPI)] could reduce the protein expression of RAGE and NOX­4. Furthermore, inhibition of RAGE led to the downregulation of vascular endothelial growth factor (VEGF) and hypoxia­inducible factor­1α (HIF­1α), thus suggesting that HG may influence angiogenesis and tumor metabolism via the RAGE­NOXs pathway. The present study also demonstrated that the RAGE­blocking antibody downregulated NOX­4 and subsequently reduced the production of downstream inflammatory factors, whereas DPI did not affect the mRNA expression of RAGE but it did reduce the protein level of RAGE and then attenuate the inflammatory response. These results indicated that inhibition of RAGE or NOXs may promote the reduced expression of VEGF and HIF­1α, and NOXs may be downstream targets of RAGE, thus indicating a HG­RAGE­NOXs­VEGF/HIF­1α association. Furthermore, the results indicated that HG may serve a role in the development of lung adenocarcinoma, mediated by the RAGE­oxidative stress pathway; therefore, the regulation of this glucose­associated pathway may be a promising novel direction for oncotherapy. However, while certain antidiabetic agents have been verified to exert inhibitory effects on tumor growth, they can also have long­term adverse effects on the body, which may limit the value of these drugs as anticancer treatments. In conclusion, the present study suggested a novel attempt to suppress glucose­induced tumor growth using a RAGE inhibitor such as soluble RAGE while avoiding the risk of glucose fluctuation.


Assuntos
Adenocarcinoma/metabolismo , Glucose/metabolismo , Neoplasias Pulmonares/metabolismo , NADPH Oxidases/metabolismo , Receptor para Produtos Finais de Glicação Avançada/metabolismo , Transdução de Sinais , Adenocarcinoma/genética , Adenocarcinoma/patologia , Adenocarcinoma de Pulmão , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células , Transformação Celular Neoplásica/genética , Transformação Celular Neoplásica/metabolismo , Humanos , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/patologia , NADPH Oxidases/antagonistas & inibidores , Metástase Neoplásica , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Receptor para Produtos Finais de Glicação Avançada/antagonistas & inibidores , Fator A de Crescimento do Endotélio Vascular/metabolismo
15.
Immunobiology ; 223(6-7): 443-448, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29398016

RESUMO

The major mite allergenic components of protease allergens (group 1,3) and non-protease allergens (group 2,7) derived from Dermatophagoides peronyssinus (Dp) and D. farinae (Df) are reported to be capable of sensitizing 80-90% of mite-allergic patients. Although protease and non-protease allergens have been demonstrated to trigger innate and adaptive immune responses through epithelium activation, the simultaneous or sequential effects of both groups of allergens has not been reported. Since all allergens are present in the mite crude extracts, it is important to determine whether these allergens can synergistically trigger the immune responses to cause airway inflammation. A total of 60 house dust mite (HDM)-allergic asthmatic patients were recruited to analyze their serum-specific IgE response to both groups of allergens. Recombinant protease allergen (Der p1 and Der p3) and non-protease allergens (Der p2 and Der p7) were used to activate the human airway epithelium cell (Beas-2B). The cells were analyzed for mRNA expression of IL-6/IL-8 and the culture supernatants were analyzed for neutrophil chemotactic activity (NCA). The results showed 48/60 (80%) HDM-allergic patients were sensitized to all allergenic components of Der p1, Der p2, Der f1, and Der f2. Most of the allergic patients were sensitized to both groups of allergens simultaneously. The associations of Der p1 with Der p2 were 83.3% (50/60) and Der f1 with Der f2 were 80% (48/60). When Beas-2B cells were cultured with Der p2 in conjunction with Der p1 and Der p3, the results showed that there was increased expression of IL-6/IL-8 in comparison with culture with allergen alone. There was only a trivial effect on IL-6/IL-8 expression when Der p2 was co-cultured with Der p7. Similar findings were obtained in the NCA measurement. When Beas-2B was cultured with Der p2 in conjunction with Der p1 and Der p3, there was increased NCA in comparison with culture with allergen alone. There were also trivial effects when Der p2 was co-cultured with Der p7. The allergens (Der p2 and Der p3)-induced IL-6/IL-8 expression and NCA released from Beas-2B could be downregulated by dexamethasone and transcription factor inhibitor SP600125. The allergenic components derived from Dp and Df can sensitize allergic patients simultaneously and activate epithelium through protease allergens (group 1, 3) and non-protease allergen (group 2) synergistically.


Assuntos
Hipersensibilidade/imunologia , Neutrófilos/imunologia , Mucosa Respiratória/imunologia , Animais , Antracenos/farmacologia , Antígenos de Dermatophagoides/imunologia , Proteínas de Artrópodes/imunologia , Células Cultivadas , Cisteína Endopeptidases/imunologia , Dermatophagoides farinae/imunologia , Dermatophagoides pteronyssinus/imunologia , Dexametasona/farmacologia , Regulação da Expressão Gênica , Humanos , Doenças do Sistema Imunitário , Imunidade Inata , Imunoglobulina E/sangue , Interleucina-6/genética , Interleucina-6/metabolismo , Interleucina-8/genética , Interleucina-8/metabolismo , Transtornos Leucocíticos , Peptídeo Hidrolases/imunologia , Mucosa Respiratória/patologia , Serina Endopeptidases/imunologia
16.
Int J Mol Sci ; 16(5): 9504-19, 2015 Apr 27.
Artigo em Inglês | MEDLINE | ID: mdl-25923080

RESUMO

The high affinity immunoglobulin E (IgE) receptor-FcεR1 is mainly expressed on the surface of effector cells. Cross-linking of IgE Abs bound to FcεR1 by multi-valent antigens can induce the activation of these cells and the secretion of inflammatory mediators. Since FcεR1 plays a central role in the induction and maintenance of allergic responses, this study aimed to investigate the association of FcεR1 with the allergic phenotype of Cε expression and cytokine and histamine release from peripheral leukocytes. Peripheral leukocytes from 67 allergic and 50 non-allergic subjects were used for genotyping analysis. Peripheral mononuclear cells (PBMCs) were used for Cε expression and ELISpot analysis, while polymorphonuclear cells (PMNs) were used for histamine release. The association between genotype polymorphism of the FcεR1α promoter region (rs2427827 and rs2251746) and allergic features of Cε expression and histamine were analyzed, and their effects on leukocytes function were compared with wild type. The genotype polymorphisms of FcεR1α promoter region with CT and TT in rs2427827 and TC in rs2251746 were significantly higher in allergic patients than in non-allergic controls. Patients with single nucleotide polymorphism (SNP) of FcεR1α promoter region had high levels of total IgE, mite-specific Der p 2 (Group 2 allergen of Dermatophagoides pteronyssinus)-specific IgE and IgE secretion B cells. The mRNA expression of FcεR1α was significantly increased after Der p2 stimulation in PBMCs with SNPs of the FcεR1α promoter region. Despite the increased Cε mRNA expression in PBMCs and histamine release from PMNs and the up-regulated mRNA expression of interleukin (IL)-6 and IL-8 secretions after Der p2 stimulation, there was no statistically significant difference between SNPs of the FcεR1α promoter region and the wild type. SNPs of FcεR1α promoter region were associated with IgE expression, IgE producing B cells, and increased Der p2-induced FcεR1α mRNA expression. These SNPs may be used as a disease marker for IgE-mediated allergic inflammation caused by Dermatophagoides pteronyssinus.


Assuntos
Hipersensibilidade Imediata/genética , Regiões Constantes de Imunoglobulina/metabolismo , Receptores de IgE/genética , Adolescente , Adulto , Alelos , Animais , Antígenos de Dermatophagoides/imunologia , Proteínas de Artrópodes/imunologia , Criança , Feminino , Genótipo , Humanos , Hipersensibilidade Imediata/imunologia , Regiões Constantes de Imunoglobulina/genética , Imunoglobulina E/biossíntese , Masculino , Pessoa de Meia-Idade , Neutrófilos/citologia , Fenótipo , Projetos Piloto , Polimorfismo Genético , Polimorfismo de Nucleotídeo Único , Regiões Promotoras Genéticas , Pyroglyphidae/imunologia , RNA Mensageiro/metabolismo , Adulto Jovem
17.
Allergy Asthma Immunol Res ; 7(4): 393-403, 2015 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-25749775

RESUMO

PURPOSE: House-dust-mite (HDM) major allergen Der p2 shares homology and function with Toll-like receptor (TLR) signaling protein myeloid differentiation-2 (MD2) and may lead to airway inflammation. Should Der p2 be internalized by human airway epithelium, it has the theoretical propensity to potentiate epithelium activation. This study aimed to demonstrate the internalization of Der p2 by airway epithelium and to investigate the effects of Der p2 on MD2 expression and epithelium activation. METHODS: Internalization of recombinant, enhanced green fluorescent protein-labelled Der p2 (rDer p2-EGFP) into human airway epithelium (BEAS-2B) was tracked by laser confocal microscopy and confirmed by immunoblotting. Reverse-transcription polymerase chain reaction (RT-PCR), immunoblotting, and immunohistochemical staining were used to determine the effect of Der p2 on MD2 expression in vitro and ex vivo. Expression of messenger RNA (mRNA) encoding receptors/cytokines was measured by RT-PCR. Secretion of interleukin-6/interleukin-8 (IL-6/IL-8) was measured by enzyme-linked immunosorbent assay (ELISA). RESULTS: Internalization of Der p2 by BEAS-2B was confirmed by confocal microscopy and immunoblotting using rDer p2-EGFP and rDer p2, respectively. Expression of MD2 protein was increased in BEAS-2B and human nasal polyp airway epithelium cultured with rDer p2. Recombinant Der p2-cultured BEAS-2B caused little spontaneous IL-6/IL-8 secretion but significantly augmented by TLR ligand LPS. IL-6 secretion was up-regulated after MD2 transfection. Internalization of Der p2 was reduced by TLR2 RNA knockdown. Dexamethasone, calcitriol, anti-MD2/anti-TLR2 antibodies, and signalling inhibitors significantly reduced LPS+Der p2-induced IL-6/IL-8 secretion. CONCLUSIONS: Human airway epithelium may internalize Der p2, which potentiates the response to environmental proinflammatory stimuli through MD2 and TLRs. This study highlights a novel mechanism and alleviates IL-6/IL-8 secretion in mite-induced airway inflammation.

18.
PLoS One ; 6(9): e23249, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-21909400

RESUMO

BACKGROUND: Allergic disease can be characterized as manifestations of an exaggerated inflammatory response to environmental allergens triggers. Mite allergen Der-p2 is one of the major allergens of the house dust mite, which contributes to TLR4 expression and function in B cells in allergic patients. However, the precise mechanisms of Der-p2 on B cells remain obscure. METHODOLOGY/PRINCIPAL FINDINGS: We investigated the effects of Der-p2 on proinflammatory cytokines responses and Toll-like receptor-4 (TLR4)-related signaling in human B cells activation. We demonstrated that Der-p2 activates pro-inflammatory cytokines, TLR4 and its co-receptor MD2. ERK inhibitor PD98059 significantly enhanced TLR4/MD2 expression in Der-p2-treated B cells. Der-p2 markedly activated mitogen-activated protein kinase (MAPK) phosphatase-1 (MKP-1) and decreased p38 phosphorylation in B cells. MKP-1-siRNA downregulated TLR4/MD2 expression in Der-p2-treated B cells. In addition, Der-p2 significantly up-regulated expression of co-stimulatory molecules and increased B cell proliferation. Neutralizing Der-p2 antibody could effectively abrogate the Der-p2-induced B cell proliferation. Der-p2 could also markedly induce NF-κB activation in B cells, which could be counteracted by dexamethasone. CONCLUSIONS/SIGNIFICANCE: These results strongly suggest that Der-p2 is capable of triggering B cell activation and MKP-1-activated p38/MAPK dephosphorylation-regulated TLR4 induction, which subsequently enhances host immune, defense responses and development of effective allergic disease therapeutics in B cells.


Assuntos
Antígenos de Dermatophagoides/imunologia , Linfócitos B/imunologia , Dermatophagoides pteronyssinus/imunologia , Ativação Linfocitária/imunologia , Ácaros/imunologia , Receptor 4 Toll-Like/imunologia , Animais , Linfócitos B/citologia , Linfócitos B/efeitos dos fármacos , Linfócitos B/enzimologia , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Citocinas/metabolismo , Dermatophagoides pteronyssinus/efeitos dos fármacos , Dexametasona/farmacologia , Fosfatase 1 de Especificidade Dupla/metabolismo , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Humanos , Mediadores da Inflamação/metabolismo , Ativação Linfocitária/efeitos dos fármacos , Antígeno 96 de Linfócito/metabolismo , Camundongos , Modelos Imunológicos , NF-kappa B/metabolismo , Fosforilação/efeitos dos fármacos , Regulação para Cima/efeitos dos fármacos , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo
19.
Eur J Pharmacol ; 590(1-3): 343-54, 2008 Aug 20.
Artigo em Inglês | MEDLINE | ID: mdl-18590725

RESUMO

Platelet-derived growth factor (PDGF) is released from vascular smooth muscle cells (VSMCs), endothelial cells, or macrophages after percutaneous coronary intervention and is related with neointimal proliferation and restenosis. Berberine is a well-known component of the Chinese herb medicine Huanglian (Coptis chinensis), and is capable of inhibiting growth and endogenous PDGF synthesis in VSMCs after in vitro mechanical injury. We analyzed the effects of berberine on VSMC growth, migration, and signaling events after exogenous PDGF stimulation in vitro in order to mimic a post-angioplasty PDGF shedding condition. Pretreatment of VSMCs with berberine inhibited PDGF-induced proliferation. Berberine significantly suppressed PDGF-stimulated Cyclin D1/D3 and Cyclin-dependent kinase (Cdk) gene expression. Moreover, berberine increased the activity of AMP-activated protein kinase (AMPK), which led to phosphorylation activation of p53 and increased protein levels of the Cdk inhibitor p21(Cip1). Compound C, an AMPK inhibitor, partly but significantly attenuated berberine-elicited growth inhibition. In addition, stimulation of VSMCs with PDGF led to a transient increase in GTP-bound, active form of Ras, Cdc42 and Rac1, as well as VSMC migration. However, pretreatment with berberine significantly inhibited PDGF-induced Ras, Cdc42 and Rac1 activation and cell migration. Co-treatment with farnesyl pyrophosphate and geranylgeranyl pyrophosphate drastically reversed berberine-mediated anti-proliferative and migratory effects in VSMCs. Based on these findings, we conclude that berberine inhibited PDGF-induced VSMC growth via activation of AMPK/p53/p21(Cip1) signaling while inactivating Ras/Rac1/Cyclin D/Cdks and suppressing PDGF-stimulated migration via inhibition of Rac1 and Cdc42. These observations offer a molecular explanation for the anti-proliferative and anti-migratory properties of berberine.


Assuntos
Berberina/farmacologia , Complexos Multienzimáticos/fisiologia , Músculo Liso Vascular/efeitos dos fármacos , Miócitos de Músculo Liso/efeitos dos fármacos , Fator de Crescimento Derivado de Plaquetas/antagonistas & inibidores , Proteínas Serina-Treonina Quinases/fisiologia , Proteínas Quinases Ativadas por AMP , Animais , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Ciclina D1/análise , Ciclina D3 , Ciclinas/análise , MAP Quinase Quinase 1/metabolismo , Músculo Liso Vascular/citologia , Ratos , Ratos Sprague-Dawley , Proteínas rac1 de Ligação ao GTP/metabolismo
20.
Apoptosis ; 11(11): 2053-63, 2006 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-17041759

RESUMO

Our previous study demonstrated that norepinephrine (NE) induces endothelial apoptosis mainly through down-regulation of Bcl-2 protein and activation of the beta-adrenergic and caspase-2 pathways. However, whether reactive oxygen species (ROS) and mitogen-activated protein kinases (MAPKs) are involved in this signal transduction remains unknown. Endothelial cells cultured from neonatal rat heart were treated with 100 microM NE. Proteins of MAPKs and Bcl-2 family were assayed by Western blotting. Apoptosis was determined by terminal deoxynucleotidyl transferase-mediated nick end-labeling assay. ROS was analyzed with flow cytometry. Caspase activity was measured using specific fluorogenic substrates. Treatment with NE increased intracellular ROS level and extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p38 phosphorylation. Whereas the phosphorylated form of Akt was decreased. The NE-induced apoptosis was abrogated by SP600125 (a specific inhibitor of JNK). Antioxidants such as vitamin C and N-acetyl cysteine inhibited NE-induced ROS production, JNK phosphorylation, caspase activation and apoptosis. Exogenously added superoxide dismutase or catalase markedly diminished NE-induced ROS production and cell death. In conclusions, our study is the first report documenting that NE induces apoptosis in neonatal rat endothelial cells via a ROS-dependent JNK activation pathway. Antioxidants may be useful in the prevention and management of NE-mediated endothelial apoptosis during heart failure.


Assuntos
Apoptose/efeitos dos fármacos , Células Endoteliais/citologia , Proteínas Quinases JNK Ativadas por Mitógeno/metabolismo , Norepinefrina/fisiologia , Espécies Reativas de Oxigênio/metabolismo , Acetilcisteína/farmacologia , Animais , Animais Recém-Nascidos , Antioxidantes/farmacologia , Ácido Ascórbico/farmacologia , Regulação para Baixo , Células Endoteliais/metabolismo , Ativação Enzimática , Peróxido de Hidrogênio/metabolismo , Quinases de Proteína Quinase Ativadas por Mitógeno/metabolismo , Miocárdio/citologia , NADPH Oxidases/metabolismo , Norepinefrina/farmacologia , Fosforilação , Proteínas Proto-Oncogênicas c-akt/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Ratos , Ratos Sprague-Dawley , Receptores Adrenérgicos/metabolismo , Transdução de Sinais
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