Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 5.463
Filtrar
1.
Sci Rep ; 11(1): 19481, 2021 09 30.
Artigo em Inglês | MEDLINE | ID: mdl-34593970

RESUMO

The pandemic infectious disease (Covid-19) caused by the coronavirus (SARS-CoV2) is spreading rapidly around the world. Covid-19 does an irreparable harm to the health and life of people. It also has a negative financial impact on the economies of most countries of the world. In this regard, the issue of creating drugs aimed at combating this disease is especially acute. In this work, molecular docking was used to study the docking of 23 compounds with QRF3a SARS-CoV2. The performed in silico modeling made it possible to identify leading compounds capable of exerting a potential inhibitory and virucidal effect. The leading compounds include chlorin (a drug used in PDT), iron(III)protoporphyrin (endogenous porphyrin), and tetraanthraquinone porphyrazine (an exogenous substance). Having taken into consideration the localization of ligands in the QRF3a SARS-CoV2, we have made an assumption about their influence on the pathogenesis of Covid-19. The interaction of chlorin, iron(III)protoporphyrin and protoporphyrin with the viral protein ORF3a were studied by fluorescence and UV-Vis spectroscopy. The obtained experimental results confirm the data of molecular docking. The results showed that a viral protein binds to endogenous porphyrins and chlorins, moreover, chlorin is a competitive ligand for endogenous porphyrins. Chlorin should be considered as a promising drug for repurposing.


Assuntos
Antivirais/química , Antivirais/metabolismo , Compostos Heterocíclicos/química , Compostos Macrocíclicos/química , Compostos Macrocíclicos/metabolismo , Proteínas Viroporinas/química , Proteínas Viroporinas/metabolismo , Sítios de Ligação , COVID-19/tratamento farmacológico , Reposicionamento de Medicamentos , Compostos Heterocíclicos/metabolismo , Ligantes , Simulação de Acoplamento Molecular , Porfirinas/química , Porfirinas/metabolismo , Protoporfirinas/química , Protoporfirinas/metabolismo , SARS-CoV-2/efeitos dos fármacos , Proteínas Viroporinas/antagonistas & inibidores
2.
Nanoscale ; 13(35): 14879-14899, 2021 Sep 17.
Artigo em Inglês | MEDLINE | ID: mdl-34533177

RESUMO

Colorectal cancer (CRC) has a poor prognosis and urgently needs better therapeutic approaches. 5-Aminolevulinic acid (ALA) induced protoporphyrin IX (PpIX) based photodynamic therapy (PDT) is already used in the clinic for several cancers but not yet well investigated for CRC. Currently, systemic administration of ALA offers a limited degree of tumour selectivity, except for intracranial tumours, limiting its wider use in the clinic. The combination of effective ALA-PDT and chemotherapy may provide a promising alternative approach for CRC treatment. Herein, theranostic Ag2S quantum dots (AS-2MPA) optically trackable in near-infrared (NIR), conjugated with endothelial growth factor receptor (EGFR) targeting Cetuximab (Cet) and loaded with ALA for PDT monotherapy or ALA/5-fluorouracil (5FU) for the combination therapy are proposed for enhanced treatment of EGFR(+) CRC. AS-2MPA-Cet exhibited excellent targeting of the high EGFR expressing cells and showed a strong intracellular signal for NIR optical detection in a comparative study performed on SW480, HCT116, and HT29 cells, which exhibit high, medium and low EGFR expression, respectively. Targeting provided enhanced uptake of the ALA loaded nanoparticles by strong EGFR expressing cells and formation of higher levels of PpIX. Cells also differ in their efficiency to convert ALA to PpIX, and SW480 was the best, followed by HT29, while HCT116 was determined as unsuitable for ALA-PDT. The therapeutic efficacy was evaluated in 2D cell cultures and 3D spheroids of SW480 and HT29 cells using AS-2MPA with either electrostatically loaded, hydrazone or amide linked ALA to achieve different levels of pH or enzyme sensitive release. Most effective phototoxicity was observed in SW480 cells using AS-2MPA-ALA-electrostatic-Cet due to enhanced uptake of the particles, fast ALA release and effective ALA-to-PpIX conversion. Targeted delivery reduced the effective ALA concentration significantly which was further reduced with codelivery of 5FU. Delivery of ALA via covalent linkages was also effective for PDT, but required a longer incubation time for the release of ALA in therapeutic doses. Phototoxicity was correlated with high levels of reactive oxygen species (ROS) and apoptotic/necrotic cell death. Hence, both AS-2MPA-ALA-Cet based PDT and AS-2MPA-ALA-Cet-5FU based chemo/PDT combination therapy coupled with strong NIR tracking of the nanoparticles demonstrate an exceptional therapeutic effect on CRC cells and excellent potential for synergistic multistage tumour targeting therapy.


Assuntos
Neoplasias Colorretais , Fotoquimioterapia , Pontos Quânticos , Ácido Aminolevulínico/farmacologia , Linhagem Celular Tumoral , Cetuximab/farmacologia , Neoplasias Colorretais/diagnóstico por imagem , Neoplasias Colorretais/tratamento farmacológico , Humanos , Imagem Óptica , Fármacos Fotossensibilizantes/farmacologia , Protoporfirinas
3.
J Photochem Photobiol B ; 223: 112286, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34416476

RESUMO

Plasmodium falciparum, the causative organism of Malaria is a mosquito-borne parasitic disease which infects red blood cells (RBCs), where it multiplies rapidly and goes through different stages of its life cycle. When the parasite load exceeds >3% in the blood, malaria transforms into severe malaria which requires immediate attention as death occurs within hours to days. The increase in people traveling to malaria-endemic areas and resistance/partial resistance to most known antimalarial drugs has put the current management scheme in jeopardy. To improve the patient outcome at this point, the physician may opt to perform exchange transfusions from another individual as an adjunct therapy to reduce parasitized RBCs, but the strategy has many drawbacks, including chances of infection. These limitations can be mitigated if the patient's own blood is withdrawn/extracted, sterilized from the parasitic load and then re-transfused almost similar to what is done in extracorporeal blood treatment for sepsis, poisoning and graft versus host disease. Thus, in the present study a light-based photochemical approach, Photodynamic Therapy (PDT) built on delta-aminolevulinic acid-protoporphyrin IX (ALA-PpIX) synthesis is exploited. This modality was effective at destruction of both resistant and susceptible strains of parasites, including at a high load mimicking severe drug resistant malaria. The current findings have set the stage for concept of an ALA-PpIX based PDT platform, "the REAP (Rapid Elimination of Active Plasmodium) strategy". This approach provides an additional tool towards the defense against multi-drug resistant severe malaria, and other intracellular blood pathogens, dependent on heme-synthesis.


Assuntos
Antimaláricos/farmacologia , Luz , Malária/patologia , Fármacos Fotossensibilizantes/farmacologia , Plasmodium falciparum/efeitos dos fármacos , Ácido Aminolevulínico/química , Antimaláricos/química , Antimaláricos/uso terapêutico , Resistência a Medicamentos/efeitos dos fármacos , Eritrócitos/parasitologia , Humanos , Cinética , Malária/tratamento farmacológico , Malária/parasitologia , Fotoquimioterapia , Fármacos Fotossensibilizantes/química , Fármacos Fotossensibilizantes/uso terapêutico , Plasmodium falciparum/isolamento & purificação , Protoporfirinas/química , Índice de Gravidade de Doença
4.
BMC Cancer ; 21(1): 971, 2021 Aug 30.
Artigo em Inglês | MEDLINE | ID: mdl-34461853

RESUMO

BACKGROUND: Photodynamic therapy with a photosensitizer such as protoporphyrin-IX, a light sensitive metabolite of heme synthesis, is a highly selective treatment for various carcinomas. In previous studies, we found a significant down regulation of the relevant enzyme ferrochelatase in gastrointestinal carcinomas leading to an accumulation of protoporphyrin-IX within the tumor cells. Recent studies showed that a novel anti-cancer drug, Alectinib, an orally available, highly selective, potent second-generation inhibitor of anaplastic lymphoma tyrosinkinase binds to ferrochelatase. Therefore, we were interested to see whether Alectinib treatment might lead to an accumulation of protoporphyrin IX. METHODS: Tumor cells of different origin were cultured, treated with LED-light and Alectinib. Results were gained by flow cytometry, immunohistochemistry and western blotting. Apoptosis was determined by flow cytometric analysis of Annexin V-FITC stained cells. In addition, cells were counterstained with propidium iodide to distinguish early apoptotic cells and late apoptotic/necrotic cells. RESULTS: Here, we report that photodynamic treatment of tumor cell lines of different origin in combination with Alectinib increased protoporphyrin-IX specific fluorescence and concomitantly cell death. CONCLUSIONS: The usage of Alectinib could be another step for enhancing the effectiveness of photodynamic therapy. Further experiments will show whether photodynamic therapy in combination with Alectinib could be a new strategy for the treatment of e.g. peritoneal disseminated carcinomas.


Assuntos
Ácido Aminolevulínico/farmacologia , Carbazóis/farmacologia , Luz , Neoplasias/tratamento farmacológico , Fotoquimioterapia/métodos , Fármacos Fotossensibilizantes/farmacologia , Piperidinas/farmacologia , Protoporfirinas/metabolismo , Fluorescência , Humanos , Neoplasias/patologia , Células Tumorais Cultivadas
5.
Nutrients ; 13(8)2021 Jul 23.
Artigo em Inglês | MEDLINE | ID: mdl-34444676

RESUMO

Iron deficiency with or without anemia, needing continuous iron supplementation, is very common in obese patients, particularly those requiring bariatric surgery. The aim of this study was to address the impact of weight loss on the rescue of iron balance in patients who underwent sleeve gastrectomy (SG), a procedure that preserves the duodenum, the main site of iron absorption. The cohort included 88 obese women; sampling of blood and duodenal biopsies of 35 patients were performed before and one year after SG. An analysis of the 35 patients consisted in evaluating iron homeostasis including hepcidin, markers of erythroid iron deficiency (soluble transferrin receptor (sTfR) and erythrocyte protoporphyrin (PPIX)), expression of duodenal iron transporters (DMT1 and ferroportin) and inflammatory markers. After surgery, sTfR and PPIX were decreased. Serum hepcidin levels were increased despite the significant reduction in inflammation. DMT1 abundance was negatively correlated with higher level of serum hepcidin. Ferroportin abundance was not modified. This study shed a new light in effective iron recovery pathways after SG involving suppression of inflammation, improvement of iron absorption, iron supply and efficiency of erythropoiesis, and finally beneficial control of iron homeostasis by hepcidin. Thus, recommendations for iron supplementation of patients after SG should take into account these new parameters of iron status assessment.


Assuntos
Gastrectomia/efeitos adversos , Hepcidinas/sangue , Ferro/deficiência , Adulto , Proteínas de Transporte de Cátions/análise , Estudos de Coortes , Suplementos Nutricionais , Duodeno/química , Duodeno/metabolismo , Eritrócitos/química , Feminino , Humanos , Absorção Intestinal/fisiologia , Ferro/administração & dosagem , Pessoa de Meia-Idade , Obesidade/sangue , Obesidade/complicações , Obesidade/cirurgia , Estudos Prospectivos , Protoporfirinas/sangue , Receptores da Transferrina/sangue , Fatores de Transcrição/análise
6.
Int J Mol Sci ; 22(16)2021 Aug 21.
Artigo em Inglês | MEDLINE | ID: mdl-34445741

RESUMO

(1) Background: coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has been linked to hematological dysfunctions, but there are little experimental data that explain this. Spike (S) and Nucleoprotein (N) proteins have been putatively associated with these dysfunctions. In this work, we analyzed the recruitment of hemoglobin (Hb) and other metabolites (hemin and protoporphyrin IX-PpIX) by SARS-Cov2 proteins using different approaches. (2) Methods: shotgun proteomics (LC-MS/MS) after affinity column adsorption identified hemin-binding SARS-CoV-2 proteins. The parallel synthesis of the peptides technique was used to study the interaction of the receptor bind domain (RBD) and N-terminal domain (NTD) of the S protein with Hb and in silico analysis to identify the binding motifs of the N protein. The plaque assay was used to investigate the inhibitory effect of Hb and the metabolites hemin and PpIX on virus adsorption and replication in Vero cells. (3) Results: the proteomic analysis by LC-MS/MS identified the S, N, M, Nsp3, and Nsp7 as putative hemin-binding proteins. Six short sequences in the RBD and 11 in the NTD of the spike were identified by microarray of peptides to interact with Hb and tree motifs in the N protein by in silico analysis to bind with heme. An inhibitory effect in vitro of Hb, hemin, and PpIX at different levels was observed. Strikingly, free Hb at 1mM suppressed viral replication (99%), and its interaction with SARS-CoV-2 was localized into the RBD region of the spike protein. (4) Conclusions: in this study, we identified that (at least) five proteins (S, N, M, Nsp3, and Nsp7) of SARS-CoV-2 recruit Hb/metabolites. The motifs of the RDB of SARS-CoV-2 spike, which binds Hb, and the sites of the heme bind-N protein were disclosed. In addition, these compounds and PpIX block the virus's adsorption and replication. Furthermore, we also identified heme-binding motifs and interaction with hemin in N protein and other structural (S and M) and non-structural (Nsp3 and Nsp7) proteins.


Assuntos
COVID-19/etiologia , Hemoglobinas/metabolismo , SARS-CoV-2/metabolismo , Proteínas não Estruturais Virais/metabolismo , Proteínas Estruturais Virais/metabolismo , COVID-19/sangue , Hemina/metabolismo , Hemoglobinas/ultraestrutura , Humanos , Simulação de Acoplamento Molecular , Ligação Proteica , Domínios Proteicos , Proteômica , Protoporfirinas/metabolismo , SARS-CoV-2/patogenicidade , Proteínas não Estruturais Virais/ultraestrutura , Proteínas Estruturais Virais/ultraestrutura , Ligação Viral , Replicação Viral
7.
Int J Mol Sci ; 22(15)2021 Jul 31.
Artigo em Inglês | MEDLINE | ID: mdl-34361019

RESUMO

Cholestatic liver diseases can progress to end-stage liver disease and reduce patients' quality of life. Although their underlying mechanisms are still incompletely elucidated, oxidative stress is considered to be a key contributor to these diseases. Heme oxygenase-1 (HO-1) is a cytoprotective enzyme that displays antioxidant action. It has been found that this enzyme plays a protective role against various inflammatory diseases. However, the role of HO-1 in cholestatic liver diseases has not yet been investigated. Here, we examined whether pharmacological induction of HO-1 by cobalt protoporphyrin (CoPP) ameliorates cholestatic liver injury. To this end, a murine model of 3,5-diethoxycarbonyl-1,4-dihydrocollidine (DDC) diet feeding was used. Administration of CoPP ameliorated liver damage and cholestasis with HO-1 upregulation in DDC diet-fed mice. Induction of HO-1 by CoPP suppressed the DDC diet-induced oxidative stress and hepatocyte apoptosis. In addition, CoPP attenuated cytokine production and inflammatory cell infiltration. Furthermore, deposition of the extracellular matrix and expression of fibrosis-related genes after DDC feeding were also decreased by CoPP. HO-1 induction decreased the number of myofibroblasts and inhibited the transforming growth factor-ß pathway. Altogether, these data suggest that the pharmacological induction of HO-1 ameliorates cholestatic liver disease by suppressing oxidative stress, hepatocyte apoptosis, and inflammation.


Assuntos
Anti-Inflamatórios/uso terapêutico , Antioxidantes/uso terapêutico , Colestase Intra-Hepática/tratamento farmacológico , Heme Oxigenase-1/metabolismo , Protoporfirinas/uso terapêutico , Animais , Anti-Inflamatórios/farmacologia , Antioxidantes/farmacologia , Apoptose , Colestase Intra-Hepática/etiologia , Colestase Intra-Hepática/metabolismo , Hepatócitos/efeitos dos fármacos , Hepatócitos/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Estresse Oxidativo , Protoporfirinas/farmacologia , Piridinas/toxicidade , Xenobióticos/toxicidade
8.
Photodiagnosis Photodyn Ther ; 35: 102452, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-34303032

RESUMO

BACKGROUND: Photodynamic therapy (PDT) is a minimally invasive cancer therapy. However, its therapeutic efficacy for prostate cancer is not yet fully understood. In this study, the predictors of therapeutic efficacy of 5-aminolevulinic acid-based PDT (ALA-PDT) on prostate cancer cells are investigated. MATERIALS AND METHODS: The human prostate cancer cell lines, PC-3, 22Rv1, DU145, and LNCap were used to investigate the effects of ALA-PDT on protoporphyrin IX (PpIX) intracellular accumulation, which was measured by flow cytometry. The cytotoxicity of ALA-PDT was evaluated by MTT (3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide) assay. The levels of porphyrin metabolism-related enzyme and transporter mRNA were comprehensively evaluated by quantitative real-time polymerase chain reaction (qRT-PCR). Protein expression was evaluated by Western blot. A xenograft model was created using PC-3 and 22Rv1, and then, pathological analysis was performed to determine the therapeutic effect of ALA-PDT RESULTS: PC-3 and LNCap cells showed high accumulation of PpIX and high sensitivity to ALA-PDT, while 22Rv1 and DU145 showed low accumulation of PpIX and low sensitivity to ALA-PDT. ALA-PDT-induced cytotoxicity correlated negatively with PpIX accumulation. The in vitro assays identified the ATP-binding cassette transporter subfamily G2 (ABCG2) transporter dimer as a predictor of treatment response. In vivo immunohistochemical staining of ABCG2 transporter showed low expression in PC-3 cells and high expression in 22Rv1 cells, and ALA-PDT-induced tumor tissue degeneration was greater in PC-3 cells than in 22Rv1 cells. CONCLUSION: The ABCG2 transporter is a useful predictor of the therapeutic effect of ALA-PDT on human prostate cancer cells.


Assuntos
Fotoquimioterapia , Neoplasias da Próstata , Ácido Aminolevulínico , Linhagem Celular Tumoral , Humanos , Masculino , Fotoquimioterapia/métodos , Fármacos Fotossensibilizantes/uso terapêutico , Neoplasias da Próstata/tratamento farmacológico , Protoporfirinas
9.
Photochem Photobiol Sci ; 20(7): 843-857, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-34216374

RESUMO

Photosensitizers of singlet oxygen exhibit three main types of reverse intersystem-crossing (RISC): thermally activated, triplet-triplet annihilation, and singlet oxygen feedback. RISC can be followed by delayed fluorescence (DF) emission, which can provide important information about the excited state dynamics in the studied system. An excellent model example is a widely used clinical photosensitizer Protoporphyrin IX, which manifests all three mentioned types of RISC and DF. Here, we estimated rate constants of individual RISC and DF processes in Protoporphyrin IX in dimethylformamide, and we showed how these affect triplet decays and DF signals under diverse experimental conditions, such as a varying oxygen concentration or excitation intensity. This provided a basis for a general discussion on guidelines for a more precise analysis of long-lived signals. Furthermore, it has been found that PpIX photoproducts and potential transient excited complexes introduce a new overlapping delayed luminescence spectral band with a distinct lifetime. These findings are important for design of more accurate biological oxygen sensors and assays based on DF and triplet lifetime.


Assuntos
Fluorescência , Hipóxia , Fármacos Fotossensibilizantes/metabolismo , Protoporfirinas/metabolismo , Humanos , Oxigênio/química , Oxigênio/metabolismo , Fármacos Fotossensibilizantes/química , Protoporfirinas/química
10.
BMC Geriatr ; 21(1): 384, 2021 06 26.
Artigo em Inglês | MEDLINE | ID: mdl-34174837

RESUMO

BACKGROUND: According to the European Working Group on Sarcopenia in Older People (EWGSOP), the diagnosis of sarcopenia primarily focused on low muscle strength with the detection of low muscle quality and quantity as confirming index. Many studies had identified mitochondrial dysfunction as one of the multifactorial etiologies of sarcopenia. Yet, no study had investigated the role of biosynthetic pathway intermediate, which was found in mitochondria, in the development of sarcopenia. This study aimed to examine the association between protoporphyrin IX (PPIX) and components of sarcopenia. METHOD: The present study enrolled 1172 participants without anemia between 1999 to 2002 from the National Health and Nutrition Examination Survey (NHANES) database. We employed the multivariable-logistic regression model to examine the relationship between PPIX and sarcopenia. Covariate adjustments were designated to each of the three models for further analysis of the relationship. RESULTS: In the unadjusted model, PPIX was significantly associated with sarcopenia (OR = 3.910, 95% CI = 2.375, 6.439, P value < 0.001). The significance persisted after covariate adjustments as observed in the fully adjusted model (OR = 2.537, 95% CI = 1.419, 4.537, P value = 0.002). CONCLUSIONS: The findings of this study suggested statistically significant association between PPIX and sarcopenia. Our study disclosed the potential of PPIX as a valuable indicator of sarcopenia.


Assuntos
Sarcopenia , Idoso , Estudos Transversais , Força da Mão , Humanos , Inquéritos Nutricionais , Prevalência , Protoporfirinas , Sarcopenia/diagnóstico , Sarcopenia/epidemiologia
11.
Int J Mol Sci ; 22(10)2021 May 14.
Artigo em Inglês | MEDLINE | ID: mdl-34069086

RESUMO

Hydrogen sulfide (H2S) is an endogenously produced molecule with anti-inflammatory and cytoprotective properties. We aimed to investigate for the first time if a novel, esterase-sensitive H2S-prodrug, BW-HS-101 with the ability to release H2S in a controllable manner, prevents gastric mucosa against acetylsalicylic acid-induced gastropathy on microscopic and molecular levels. Wistar rats were pretreated intragastrically with vehicle, BW-HS-101 (0.5-50 µmol/kg) or its analogue without the ability to release H2S, BW-iHS-101 prior to ASA administration (125 mg/kg, intragastrically). BW-HS-101 was administered alone or in combination with nitroarginine (L-NNA, 20 mg/kg, intraperitoneally) or zinc protoporphyrin IX (10 mg/kg, intraperitoneally). Gastroprotective effects of BW-HS-101 were additionally evaluated against necrotic damage induced by intragastrical administration of 75% ethanol. Gastric mucosal damage was assessed microscopically, and gastric blood flow was determined by laser flowmetry. Gastric mucosal DNA oxidation and PGE2 concentration were assessed by ELISA. Serum and/or gastric protein concentrations of IL-1α, IL-1ß, IL-2, IL-4, IL-6, IL-10, IL-13, VEGF, GM-CSF, IFN-γ, TNF-α, and EGF were determined by a microbeads/fluorescent-based multiplex assay. Changes in gastric mucosal iNOS, HMOX-1, SOCS3, IL1-R1, IL1-R2, TNF-R2, COX-1, and COX-2 mRNA were assessed by real-time PCR. BW-HS-101 or BW-iHS-101 applied at a dose of 50 µmol/kg protected gastric mucosa against ASA-induced gastric damage and prevented a decrease in the gastric blood flow level. H2S prodrug decreased DNA oxidation, systemic and gastric mucosal inflammation with accompanied upregulation of SOCS3, and EGF and HMOX-1 expression. Pharmacological inhibition of nitric oxide (NO) synthase but not carbon monoxide (CO)/heme oxygenase (HMOX) activity by L-NNA or ZnPP, respectively, reversed the gastroprotective effect of BW-HS-101. BW-HS-101 also protected against ethanol-induced gastric injury formation. We conclude that BW-HS-101, due to its ability to release H2S in a controllable manner, prevents gastric mucosa against drugs-induced gastropathy, inflammation and DNA oxidation, and upregulate gastric microcirculation. Gastroprotective effects of this H2S prodrug involves endogenous NO but not CO activity and could be mediated by cytoprotective and anti-inflammatory SOCS3 and EGF pathways.


Assuntos
Mucosa Gástrica/efeitos dos fármacos , Sulfeto de Hidrogênio/farmacocinética , Substâncias Protetoras/farmacologia , Animais , Anti-Inflamatórios não Esteroides/efeitos adversos , Aspirina/efeitos adversos , DNA/metabolismo , Liberação Controlada de Fármacos , Etanol/toxicidade , Mucosa Gástrica/irrigação sanguínea , Mucosa Gástrica/patologia , Gastrite/induzido quimicamente , Gastrite/tratamento farmacológico , Gastrite/patologia , Regulação da Expressão Gênica/efeitos dos fármacos , Masculino , Óxido Nítrico/metabolismo , Nitroarginina/administração & dosagem , Nitroarginina/farmacologia , Pró-Fármacos/farmacocinética , Prostaglandina-Endoperóxido Sintases/metabolismo , Prostaglandinas/metabolismo , Substâncias Protetoras/administração & dosagem , Protoporfirinas/administração & dosagem , Protoporfirinas/farmacologia , Ratos Wistar
12.
Methods Mol Biol ; 2277: 175-185, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34080152

RESUMO

The Protoporphyrin IX-Triplet State Lifetime Technique (PpIX-TSLT) has been proposed by us as a potential clinical noninvasive tool for monitoring mitochondrial function. We have been working on the development of mitochondrial respirometry for monitoring mitochondrial oxygen tension (mitoPO2) and mitochondrial oxygen consumption (mitoVO2) in skin. In this work, we describe the principles of the method in small experimental animals.


Assuntos
Mitocôndrias/metabolismo , Consumo de Oxigênio , Ácido Aminolevulínico/farmacologia , Animais , Temperatura Corporal , Desenho de Equipamento , Medições Luminescentes/instrumentação , Medições Luminescentes/métodos , Protoporfirinas/química , Ratos Wistar , Respiração Artificial , Pele/efeitos dos fármacos , Traqueotomia
13.
Molecules ; 26(11)2021 May 28.
Artigo em Inglês | MEDLINE | ID: mdl-34071361

RESUMO

Nuclear receptor REV-ERBß is an overexpressed oncoprotein that has been used as a target for cancer treatment. The metal-complex nature of its ligand, iron protoporphyrin IX (Heme), enables the REV-ERBß to be used for multiple therapeutic modalities as a photonuclease, a photosensitizer, or a fluorescence imaging agent. The replacement of iron with cobalt as the metal center of protoporphyrin IX changes the ligand from an agonist to an antagonist of REV-ERBß. The mechanism behind that phenomenon is still unclear, despite the availability of crystal structures of REV-ERBß in complex with Heme and cobalt protoporphyrin IX (CoPP). This study used molecular dynamic simulations to compare the effects of REV-ERBß binding to Heme and CoPP, respectively. The initial poses of Heme and CoPP in complex with agonist and antagonist forms of REV-ERBß were predicted using molecular docking. The binding energies of each ligand were calculated using the MM/PBSA method. The computed binding affinity of Heme to REV-ERBß was stronger than that of CoPP, in agreement with experimental results. CoPP altered the conformation of the ligand-binding site of REV-ERBß, disrupting the binding site for nuclear receptor corepressor, which is required for REV-ERBß to regulate the transcription of downstream target genes. Those results suggest that a subtle change in the metal center of porphyrin can change the behavior of porphyrin in cancer cell signaling. Therefore, modification of porphyrin-based agents for cancer therapy should be conducted carefully to avoid triggering unfavorable effects.


Assuntos
Cobalto/química , Neoplasias/tratamento farmacológico , Protoporfirinas/antagonistas & inibidores , Receptores Citoplasmáticos e Nucleares/química , Proteínas Repressoras/química , Sítios de Ligação , Química Farmacêutica/métodos , Heme/química , Humanos , Ferro/química , Cinética , Ligantes , Metais , Conformação Molecular , Simulação de Acoplamento Molecular , Simulação de Dinâmica Molecular , Peptídeos/química , Fármacos Fotossensibilizantes/química , Porfirinas/química , Ligação Proteica , Receptores Citoplasmáticos e Nucleares/antagonistas & inibidores , Receptores Citoplasmáticos e Nucleares/metabolismo , Proteínas Repressoras/antagonistas & inibidores , Proteínas Repressoras/genética , Transdução de Sinais
14.
Int J Pharm ; 605: 120813, 2021 Aug 10.
Artigo em Inglês | MEDLINE | ID: mdl-34144137

RESUMO

Heme is a prosthetic group of hemoglobin comprising protoporphyrin IX (PPIX) with Fe2+. Studies have shown that modulating heme synthesis pathway in Plasmodium could greatly affect the action mechanism and antimalarial effect of artemisinin and its derivatives. Herein, an intraerythrocytic parasite targeted nanostructured lipid carrier (NLC) was developed for potentiation of artemether (ARM) by combination with PPIX and iron-loaded transferrin (holo-Tf). Firstly, ARM and PPIX were co-loaded into NLCs with high entrapment efficiency. Then, a targeting ligand heparin (HP) was electrostatically adsorbed onto the periphery of NLCs, followed by conjugation with holo-Tf to receive the final formulation Tf-HP-NLC/ARM/PPIX. Tf-HP-NLC/ARM/PPIX exhibited nanoscale particle size (~188 nm) and was relatively stable in simulated gastrointestinal fluids and rat plasma. A sustained drug release characteristic was observed in PBS (pH 7.4). In vitro targeting assay confirmed that Tf-HP-NLC/ARM/PPIX could be specifically and efficiently internalized into intraerythrocytic parasites via HP receptor-meditated endocytosis. Furthermore, due to enhanced intraparasitic accumulation and activated mechanism of ARM, the combinational delivery system Tf-HP-NLC/ARM/PPIX showed increased inhibitory activity against Plasmodium falciparum in culture and enhanced antimalarial effect in Plasmodium berghei-infected murine model, suggesting a promising strategy for development of new therapy based on action mechanism of ARM.


Assuntos
Malária , Nanoestruturas , Animais , Artemeter/uso terapêutico , Portadores de Fármacos/uso terapêutico , Heparina , Lipídeos , Malária/tratamento farmacológico , Camundongos , Tamanho da Partícula , Protoporfirinas , Ratos , Transferrina
15.
ACS Appl Mater Interfaces ; 13(24): 27991-27998, 2021 Jun 23.
Artigo em Inglês | MEDLINE | ID: mdl-34110123

RESUMO

Porphyria is a group of genetic photodermatoses that cause too much porphyrin to accumulate in the blood, skin, and liver, resulting in skin photosensitivity and damage, liver disease, or potential liver failure. Conventional detection methods include high-performance liquid chromatography and fluorescence spectrometry. However, these methods usually require complicated pretreatment and time-consuming processes. Therefore, efficient and fast detection of porphyria is urgently needed. Herein, we develop a molecular afterglow reporter-based sensing scheme for the detection of porphyrins in whole blood. The afterglow reporter can respond to the production of singlet oxygen (1O2) of porphyrins after light excitation, and the detection signals can be amplified through adjusting the amount of singlet oxygen and afterglow reporter molecules. Moreover, without the use of a real-time excitation source, afterglow signals can avoid the scattering and autofluorescence interference in biological samples, thereby reducing background noise. More importantly, we prove the applicability of the afterglow reporter in the quantitative detection of porphyrins in whole blood and demonstrate its great clinical potential.


Assuntos
Adamantano/análogos & derivados , Corantes Fluorescentes/química , Porfirias/diagnóstico , Protoporfirinas/análise , Quinolinas/química , Espectrometria de Fluorescência/métodos , Adamantano/síntese química , Corantes Fluorescentes/síntese química , Humanos , Limite de Detecção , Porfirias/sangue , Protoporfirinas/efeitos da radiação , Quinolinas/síntese química , Oxigênio Singlete/metabolismo
16.
Arch Biochem Biophys ; 706: 108926, 2021 07 30.
Artigo em Inglês | MEDLINE | ID: mdl-34029560

RESUMO

Morroniside, a major iridoid glycoside isolated from Cornus officinalis, has a variety of beneficial pharmacological properties. Although morroniside has recently been reported to exhibit anti-inflammatory and antioxidant effects, the detailed mechanism has not yet been fully elucidated. In this study, we investigated the inhibitory effect of morroniside on inflammatory and oxidative stress activated by lipopolysaccharide (LPS) in RAW 264.7 macrophages. Our results indicated that morroniside pretreatment significantly inhibited the LPS-induced phagocytic activity and release of pro-inflammatory factors, which was associated with blocking the expression of their regulatory genes. Morroniside also markedly suppressed the expression of myeloid differentiation factor 88 as well as Toll-like receptor 4 (TLR4), and attenuated the translocation of nuclear factor-κB (NF-κB) to the nucleus in LPS-treated RAW 264.7 macrophages. Furthermore, morroniside prevented the binding of LPS to the TLR4 on the cell surface. In addition, morroniside abolished reactive oxygen species (ROS) generation, and enhanced the expression of heme oxygenase-1 (HO-1) following activation of nuclear factor-E2-related factor 2 (Nrf2) in LPS-stimulated RAW 264.7 macrophages. However, zinc protoporphyrin, a specific inhibitor of HO-1, reversed the morroniside-mediated inhibition of inflammatory response in LPS-treated RAW 264.7 macrophages. In conclusion, our findings suggest that morroniside exerts LPS-induced anti-inflammatory and antioxidant effects by targeting the TLR4/NF-κB and Nrf2/HO-1 signaling pathways in RAW 264.7 macrophages. Taken together, our findings suggest that morroniside interacted structurally and electrochemically with TLR4/MD2 complex, consequently can be a potential functional agent to prevent inflammatory and oxidative damage.


Assuntos
Anti-Inflamatórios/farmacologia , Antioxidantes/farmacologia , Glicosídeos/farmacologia , Heme Oxigenase-1/genética , Proteínas de Membrana/genética , Fator 2 Relacionado a NF-E2/genética , NF-kappa B/genética , Receptor 4 Toll-Like/genética , Animais , Anti-Inflamatórios/isolamento & purificação , Antioxidantes/isolamento & purificação , Cornus/química , Regulação da Expressão Gênica , Glicosídeos/isolamento & purificação , Heme Oxigenase-1/metabolismo , Lipopolissacarídeos/antagonistas & inibidores , Lipopolissacarídeos/farmacologia , Proteínas de Membrana/metabolismo , Camundongos , Fator 88 de Diferenciação Mieloide/genética , Fator 88 de Diferenciação Mieloide/metabolismo , Fator 2 Relacionado a NF-E2/metabolismo , NF-kappa B/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Extratos Vegetais/química , Protoporfirinas/farmacologia , Células RAW 264.7 , Espécies Reativas de Oxigênio/antagonistas & inibidores , Espécies Reativas de Oxigênio/metabolismo , Transdução de Sinais , Receptor 4 Toll-Like/metabolismo
17.
Photodiagnosis Photodyn Ther ; 34: 102327, 2021 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-33962057

RESUMO

BACKGROUND: Photodynamic therapy (PDT) has been shown to be less effective on the extremities. Protoporphyrin-IX (PpIX) fluorescence and skin surface temperature are variables that have been implicated in the differences in efficacy with body site, but objective studies have not been undertaken. OBJECTIVES: To further investigate observations from our previous study that temperature and fluorescence during pro-drug incubation are correlated, through a prospective objective investigation of the relationships between fluorescence and skin surface temperature before and during PDT and relationships with body site and efficacy. METHODS: Eighteen patients with Bowen's disease or basal cell carcinoma, who had been referred for PDT, were recruited to this study. PpIX fluorescence and thermal measurements were recorded at intervals during the pro-drug incubation and irradiation phases of PDT. Pain immediately after irradiation, and outcome at 3- and 12-months were recorded. RESULTS: Temperature and PpIX fluorescence were higher on the trunk than lower leg immediately before treatment (median temperature 32.7 °C vs. 27.8 °C, p < 0.05 and median fluorescence 16.5 vs. 6.7, p < 0.05). Higher pain levels were reported during PDT on the extremities (median 5.7 vs. 2.2, p < 0.05). Clearance rates at 12-months were 80 %. CONCLUSIONS: The study supports a correlation between temperature and PpIX fluorescence during PDT, providing robust objective data to support our previous hypothesis and observations. The higher pain levels, lower PpIX fluorescence on the lower leg, and the high efficacy rates at all body sites irrespective of temperature and fluorescence indicates that relationships between PDT treatment conditions and parameters is likely to be multifactorial.


Assuntos
Fotoquimioterapia , Neoplasias Cutâneas , Ácido Aminolevulínico/uso terapêutico , Fluorescência , Humanos , Fotoquimioterapia/métodos , Fármacos Fotossensibilizantes/uso terapêutico , Estudos Prospectivos , Protoporfirinas/uso terapêutico , Pele , Neoplasias Cutâneas/tratamento farmacológico
18.
Photodiagnosis Photodyn Ther ; 34: 102324, 2021 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-33965601

RESUMO

BACKGROUND: SARS-CoV-2 attacks hemoglobin through its structural protein ORF3a, dissociating the iron from the heme, as iron is necessary by cell machinery for virus replication. In this process protoporphyrin (PpIX) is released. METHODS: The decrease in the hemoglobin levels observed in patients with Covid-19 is frequently accompanied by an increase in PpIX levels. This evidence was confirmed by the quantification of PpIX by high-performance liquid chromatography (HPLC). PpIX emission is observed in its two characteristic bands at approximately 635 nm and 705 nm. RESULTS: This paper searches to understand the role of heme and PpIX inside the cells. Perspectives on the use of PpIX fluorescence as a sensor to monitor the presence of SARS-CoV-2 in the tissue, blood, urine, or feces to map the evolution and severity of the disease or to monitor the response of the Covid-19 treatment modalities were described. CONCLUSION: Fluorescence spectroscopy could be adopted as an excellent diagnostic technique for Covid-19, of low cost and high sensitivity. This method can potentially be used as a marker to monitor the response to the treatments. Photodynamic and sonodynamic therapies using the endogenous PpIX increased in the acute phase of the disease, could be employed for Covid-19 treatment.


Assuntos
COVID-19 , Fotoquimioterapia , Ácido Aminolevulínico , COVID-19/tratamento farmacológico , Hemoglobinas , Humanos , Fotoquimioterapia/métodos , Fármacos Fotossensibilizantes/farmacologia , Protoporfirinas , SARS-CoV-2
19.
Photodiagnosis Photodyn Ther ; 35: 102317, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-33940210

RESUMO

INTRODUCTION: Nanoparticles (Np) can increase drug efficacy and overcome problems associated with solubility and aggregation in a solution of PpIX. PURPOSE: Evaluate if Np interferes in the photophysical and photobiological capacity of the PpIX comparing with free PpIX intended for topical PDT of melanoma. METHODS: In vitro photophysical evaluation of Np-PpIX was carried out through singlet oxygen (1O2) quantum yield. In vitro cytotoxicity and phototoxicity assays have used murine melanoma cell culture. RESULTS: The quantum yield of singlet oxygen has shown that Np did not influence the formation capacity of this reactive species. In the dark, all PpIX-Nps concentrations were less cytotoxic compared to free drugs. At a higher light dose (1500 mJ.cm2) 3.91 µg / mL PpIX had similar % viable cells for free and Np (∼34 %) meaning Nps did not interfere in the photodynamic effect of PpIX. However, at 7.91 µg / mL the phototoxicity increased for both (5.8 % viable cells for free versus 21.7 % for Nps). Despite the higher phototoxicity of free PpIX at this concentration, greater cytotoxicity in the dark was obtained (∼49 % viable cells for free versus ∼90.6 % Np) which means Nps protect the tumor tissue from the photodynamic action of PpIX. CONCLUSIONS: Np is a potential delivery system for melanoma skin cancer, since it maintained the photophysical properties of PpIX and excellent in vitro phototoxicity effect against melanoma cells, reducing cell viability ∼80 % (7.91 µg / mL PpIX in Nps) and provides safe PDT (due to lower cytotoxicity in the dark).


Assuntos
Melanoma , Nanopartículas , Fotoquimioterapia , Animais , Melanoma/tratamento farmacológico , Camundongos , Fotoquimioterapia/métodos , Fármacos Fotossensibilizantes/farmacologia , Protoporfirinas
20.
J Biol Chem ; 297(1): 100778, 2021 07.
Artigo em Inglês | MEDLINE | ID: mdl-34023387

RESUMO

Porphyrias are rare blood disorders caused by genetic defects in the heme biosynthetic pathway and are associated with the accumulation of high levels of porphyrins that become cytotoxic. Porphyrins, due to their amphipathic nature, spontaneously associate into different nanostructures, but very little is known about the cytotoxic effects of these porphyrin nanostructures. Previously, we demonstrated the unique ability of fluorescent biological porphyrins, including protoporphyrin-IX (PP-IX), to cause organelle-selective protein aggregation, which we posited to be a major mechanism by which fluorescent porphyrins exerts their cytotoxic effect. Herein, we tested the hypothesis that PP-IX-mediated protein aggregation is modulated by different PP-IX nanostructures via a mechanism that depends on their oxidizing potential and protein-binding ability. UV-visible spectrophotometry showed pH-mediated reversible transformations of PP-IX nanostructures. Biochemical analysis showed that PP-IX nanostructure size modulated PP-IX-induced protein oxidation and protein aggregation. Furthermore, albumin, the most abundant serum protein, preferentially binds PP-IX dimers and enhances their oxidizing ability. PP-IX binding quenched albumin intrinsic fluorescence and oxidized His-91 residue to Asn/Asp, likely via a previously described photo-oxidation mechanism for other proteins. Extracellular albumin protected from intracellular porphyrinogenic stress and protein aggregation by acting as a PP-IX sponge. This work highlights the importance of PP-IX nanostructures in the context of porphyrias and offers insights into potential novel therapeutic approaches.


Assuntos
Nanoestruturas/química , Agregados Proteicos , Protoporfirinas/química , Ácido Aminolevulínico/farmacologia , Animais , Sítios de Ligação , Bovinos , Linhagem Celular , Desferroxamina/farmacologia , Dimerização , Humanos , Concentração de Íons de Hidrogênio , Modelos Biológicos , Oxirredução , Ligação Proteica , Estrutura Secundária de Proteína , Soroalbumina Bovina/química , Soroalbumina Bovina/metabolismo , Espectrometria de Fluorescência , Espectrofotometria Ultravioleta
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...