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1.
Shanghai Kou Qiang Yi Xue ; 29(4): 359-364, 2020 Aug.
Artigo em Chinês | MEDLINE | ID: mdl-33089282

RESUMO

PURPOSE: The aim of the present study was to investigate the effect of curcumin (Cur) on TGF-ß1/Smad3 pathway of rat gingival fibroblast treated with cyclosporine A (CsA) in vitro, and to provide theoretical basis for the mechanism of curcumin inhibiting drug-induced gingival hyperplasia induced by CsA. METHODS: Healthy Sprague-Dawley rat gingival fibroblasts were cultured with different concentrations of Cur (0, 5, 10, 20, 30 µmol/L) and Cur (20 µmol/L)+CsA(200 ng/mL), cell proliferation was assessed with CCK-8 assay. The mRNA levels of TGF-ß1, Smad3, α-SMA and collagen type Ⅰ in gingival fibroblasts were detected by real-time PCR under Cur(20 µmol/L)+CsA(200 ng/mL); the protein level of TGF-ß1, Smad3, p-Smad3, α-SMA and collagen type Ⅰ were determined through Western blot. The effect of Cur(20 µmol/L)+CsA(200 ng/mL) on migration ability of gingival fibroblasts was observed through Scratch wound-healing assay. The data were analyzed with SPSS 23.0 software package. RESULTS: Cell proliferation and migration ability of rats gingival fibroblasts were significantly reduced under Cur(20 µmol/L)+CsA(200 ng/mL). 20 µmol/L Cur significantly decreased mRNA expression of TGF-ß1, α-SMA and collagen type Ⅰ in gingival fibroblasts, and Western blot suggested significantly down-regulated expression of TGF-ß1, p-Smad3, α-SMA, and collagen typeⅠ. CONCLUSIONS: Cur may inhibit TGF-ß1/Smad3 signaling pathway of gingival fibroblasts activated by CsA, thereby weakening proliferation and migration, reducing secretion of smooth muscle actin and collagen of gingival fibroblasts, and ameliorating gingival hyperplasia.


Assuntos
Curcumina , Fator de Crescimento Transformador beta1 , Animais , Curcumina/farmacologia , Ciclosporina/farmacologia , Fibroblastos , Ratos , Ratos Sprague-Dawley
2.
Chem Biol Interact ; 330: 109245, 2020 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-32866465

RESUMO

The calcineurin inhibitor, cyclosporin A (CsA) is one of the most common immunosuppressive agents used in organ transplantation. However, its clinical use is often limited by several unwanted effects including nephrotoxicity and hepatotoxicity. By using immunohistochemical and ELISA techniques, it was found that CsA administration causes a rapid activation of a disintegrin and metalloproteases-17 (ADAM-17), epidermal growth factor receptor (EGFR) and subsequent ERK1/2 phosphorylation in the liver and kidney of albino mice. Furthermore, this study presents mechanistic relevance of this signaling cascade involving reactive oxygen species (ROS)-mediated ADAM-17/EGFR/ERK1/2 activation as indicated by a clear reduction in ADAM-17 and EGFR activities as well as ERK1/2 phosphorylation when the animals pretreated with Polyethylene glycol-superoxide dismutase (PEG-SOD) before CsA administration. Collectively, our findings demonstrate that CsA has the ability to activate ADAM-17-mediated EGFR/ERK1/2 phosphorylation in the liver and kidney of albino mice in ROS-dependent manner. Finally, these data may support the concept of using antioxidant therapy as a valuable approach for the prevention of CsA-induced nephrotoxicity and hepatotoxicity.


Assuntos
Ciclosporina/toxicidade , Rim/metabolismo , Fígado/metabolismo , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Polietilenoglicóis/farmacologia , Superóxido Dismutase/farmacologia , Proteína ADAM17/metabolismo , Animais , Ciclosporina/farmacologia , Interações Medicamentosas , Receptores ErbB/metabolismo , Rim/efeitos dos fármacos , Fígado/efeitos dos fármacos , Camundongos , Fosforilação/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo
3.
PLoS One ; 15(9): e0237809, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32915792

RESUMO

Chimeric mice with humanized livers are considered a useful animal model for predicting human (h-) drug metabolism and toxicity. In this study, the characteristics of fresh h-hepatocytes (cFHHs, PXB-cells®) isolated from chimeric mice (PXB-mice®) were evaluated in vitro to confirm their utility for drug development. cFHHs cultured at high density (2.13 × 105 cells/cm2) displayed stable production of h-albumin and cytochrome P450 (CYP) 3A activities for at least 21 days. The mRNA expression levels of 10 of 13 CYP, UDP-glucuronosyltransferase (UGT), and transporters were maintained at >10% of the levels of freshly isolated cFHHs after 21 days. From 1 week, many bile canaliculi were observed between cFHHs, and the accumulation of the multidrug resistance-associated protein and bile salt export pump substrates in these bile canaliculi was clearly inhibited by cyclosporin A. Microarray analysis of cFHHs cultured at high density and at low density (0.53 × 105 cells/cm2) revealed that high density culture maintained high expressions of some transcription factors (HNF4α, PXR, and FXR) perhaps involved in the high CYP, UGT and transporter gene expressions of cFHHs. These results strongly suggest that cFHHs could be a novel in vitro tool for drug development studies.


Assuntos
Canalículos Biliares/efeitos dos fármacos , Desenvolvimento de Medicamentos/métodos , Hepatócitos/efeitos dos fármacos , Cultura Primária de Células/métodos , Quimeras de Transplante , Animais , Canalículos Biliares/citologia , Canalículos Biliares/metabolismo , Células Cultivadas , Criança , Pré-Escolar , Ciclosporina/farmacologia , Citocromo P-450 CYP3A/genética , Citocromo P-450 CYP3A/metabolismo , Feminino , Glucuronosiltransferase/genética , Glucuronosiltransferase/metabolismo , Hepatócitos/citologia , Hepatócitos/metabolismo , Humanos , Masculino , Camundongos , Camundongos SCID , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo
4.
Antiviral Res ; 181: 104878, 2020 09.
Artigo em Inglês | MEDLINE | ID: mdl-32679055

RESUMO

In response to the current pandemic caused by the novel SARS-CoV-2, identifying and validating effective therapeutic strategies is more than ever necessary. We evaluated the in vitro antiviral activities of a shortlist of compounds, known for their cellular broad-spectrum activities, together with drugs that are currently under evaluation in clinical trials for COVID-19 patients. We report the antiviral effect of remdesivir, lopinavir, chloroquine, umifenovir, berberine and cyclosporine A in Vero E6 cells model of SARS-CoV-2 infection, with estimated 50% inhibitory concentrations of 0.99, 5.2, 1.38, 3.5, 10.6 and 3 µM, respectively. Virus-directed plus host-directed drug combinations were also investigated. We report a strong antagonism between remdesivir and berberine, in contrast with remdesivir/diltiazem, for which we describe high levels of synergy, with mean Loewe synergy scores of 12 and peak values above 50. Combination of host-directed drugs with direct acting antivirals underscore further validation in more physiological models, yet they open up interesting avenues for the treatment of COVID-19.


Assuntos
Antivirais/farmacologia , Betacoronavirus/efeitos dos fármacos , Infecções por Coronavirus/tratamento farmacológico , Reposicionamento de Medicamentos , Pandemias , Pneumonia Viral/tratamento farmacológico , Monofosfato de Adenosina/análogos & derivados , Monofosfato de Adenosina/farmacologia , Alanina/análogos & derivados , Alanina/farmacologia , Animais , Berberina/farmacologia , Chlorocebus aethiops , Cloroquina/farmacologia , Infecções por Coronavirus/virologia , Ciclosporina/farmacologia , Antagonismo de Drogas , Combinação de Medicamentos , Sinergismo Farmacológico , Humanos , Indóis/farmacologia , Lopinavir/farmacologia , Pneumonia Viral/virologia , Células Vero
5.
PLoS One ; 15(7): e0224952, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32692785

RESUMO

Tauopathies are a class of neurodegenerative disorders characterized by abnormal deposition of post-translationally modified tau protein in the human brain. Tauopathies are associated with Alzheimer's disease (AD), chronic traumatic encephalopathy (CTE), and other diseases. Hyperphosphorylation increases tau tendency to aggregate and form neurofibrillary tangles (NFT), a pathological hallmark of AD. In this study, okadaic acid (OA, 100 nM), a protein phosphatase 1/2A inhibitor, was treated for 24h in mouse neuroblastoma (N2a) and differentiated rat primary neuronal cortical cell cultures (CTX) to induce tau-hyperphosphorylation and oligomerization as a cell-based tauopathy model. Following the treatments, the effectiveness of different kinase inhibitors was assessed using the tauopathy-relevant tau antibodies through tau-immunoblotting, including the sites: pSer202/pThr205 (AT8), pThr181 (AT270), pSer202 (CP13), pSer396/pSer404 (PHF-1), and pThr231 (RZ3). OA-treated samples induced tau phosphorylation and oligomerization at all tested epitopes, forming a monomeric band (46-67 kDa) and oligomeric bands (170 kDa and 240 kDa). We found that TBB (a casein kinase II inhibitor), AR and LiCl (GSK-3 inhibitors), cyclosporin A (calcineurin inhibitor), and Saracatinib (Fyn kinase inhibitor) caused robust inhibition of OA-induced monomeric and oligomeric p-tau in both N2a and CTX culture. Additionally, a cyclin-dependent kinase 5 inhibitor (Roscovitine) and a calcium chelator (EGTA) showed contrasting results between the two neuronal cultures. This study provides a comprehensive view of potential drug candidates (TBB, CsA, AR, and Saracatinib), and their efficacy against tau hyperphosphorylation and oligomerization processes. These findings warrant further experimentation, possibly including animal models of tauopathies, which may provide a putative Neurotherapy for AD, CTE, and other forms of tauopathy-induced neurodegenerative diseases.


Assuntos
Quinase 3 da Glicogênio Sintase/antagonistas & inibidores , Ácido Okadáico/farmacologia , Inibidores de Proteínas Quinases/farmacologia , Multimerização Proteica/efeitos dos fármacos , Animais , Linhagem Celular , Ciclosporina/farmacologia , Quinase 3 da Glicogênio Sintase/metabolismo , Cloreto de Lítio/farmacologia , Camundongos , Modelos Biológicos , Fosforilação/efeitos dos fármacos , Ratos , Tauopatias/metabolismo , Tauopatias/patologia , Triazóis/farmacologia
6.
Nat Commun ; 11(1): 3347, 2020 07 03.
Artigo em Inglês | MEDLINE | ID: mdl-32620768

RESUMO

A sharp increase in mitochondrial Ca2+ marks the activation of brown adipose tissue (BAT) thermogenesis, yet the mechanisms preventing Ca2+ deleterious effects are poorly understood. Here, we show that adrenergic stimulation of BAT activates a PKA-dependent mitochondrial Ca2+ extrusion via the mitochondrial Na+/Ca2+ exchanger, NCLX. Adrenergic stimulation of NCLX-null brown adipocytes (BA) induces a profound mitochondrial Ca2+ overload and impaired uncoupled respiration. Core body temperature, PET imaging of glucose uptake and VO2 measurements confirm a thermogenic defect in NCLX-null mice. We show that Ca2+ overload induced by adrenergic stimulation of NCLX-null BAT, triggers the mitochondrial permeability transition pore (mPTP) opening, leading to a remarkable mitochondrial swelling and cell death. Treatment with mPTP inhibitors rescue mitochondrial function and thermogenesis in NCLX-null BAT, while calcium overload persists. Our findings identify a key pathway through which BA evade apoptosis during adrenergic stimulation of uncoupling. NCLX deletion transforms the adrenergic pathway responsible for thermogenesis activation into a death pathway.


Assuntos
Adipócitos Marrons/patologia , Tecido Adiposo Marrom/metabolismo , Norepinefrina/metabolismo , Trocador de Sódio e Cálcio/metabolismo , Termogênese/fisiologia , Adipócitos Marrons/citologia , Adipócitos Marrons/efeitos dos fármacos , Tecido Adiposo Marrom/citologia , Adrenérgicos/farmacologia , Animais , Apoptose/efeitos dos fármacos , Cálcio/metabolismo , Células Cultivadas , Temperatura Baixa/efeitos adversos , Ciclosporina/farmacologia , Metabolismo Energético/efeitos dos fármacos , Metabolismo Energético/fisiologia , Feminino , Microscopia Intravital , Masculino , Camundongos , Camundongos Knockout , Microscopia de Fluorescência , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Proteínas de Transporte da Membrana Mitocondrial/antagonistas & inibidores , Proteínas de Transporte da Membrana Mitocondrial/metabolismo , Cultura Primária de Células , Transdução de Sinais , Trocador de Sódio e Cálcio/genética , Termogênese/efeitos dos fármacos
7.
Exp Hematol ; 86: 21-27.e2, 2020 06.
Artigo em Inglês | MEDLINE | ID: mdl-32437909

RESUMO

Our previous study revealed that expression of G protein-coupled receptor 68 (GPR68) was upregulated in MDSL cells, a cell line representing myelodysplastic syndromes (MDS), in response to lenalidomide (LEN), and mediated a calcium/calpain proapoptotic pathway. Isx, a GPR68 agonist, enhanced the sensitivity to LEN in MDSL cells. The fact that Isx is not a U.S. Food and Drug Administration-approved drug prompts us to look for alternative candidates that could enhance the sensitivity of LEN in MDS as well as other hematologic malignancies, such as acute myeloid leukemia (AML). In the study described here, we found that regulator of calcineurin 1 (RCAN1), an endogenous inhibitor of calcineurin (CaN), was upregulated in MDSL cells in response to LEN, possibly through degradation of IKZF1. Consistently, cyclosporin (Cys), a pharmacological inhibitor of CaN, inhibited the activity of CaN and induced apoptosis in MDSL cells, indicating that CaN provided a prosurvival signal in MDSL cells. In addition, Cys enhanced the cytotoxic effect of LEN in MDS/AML cell lines as well as primary bone marrow cells from MDS patients and AML patient-derived xenograft models. Intriguingly, pretreatment with LEN reversed the suppressive effect of Cys on T-cell activation. Our study suggests a novel mechanism of action of LEN in mediating cytotoxicity in MDS/AML via upregulation of RCAN1, thus inhibiting the CaN prosurvival pathway. Our study also suggests that Cys enhances the sensitivity to LEN in MDS/AML cells without compromising T-cell activation.


Assuntos
Ciclosporina/farmacologia , Resistencia a Medicamentos Antineoplásicos/efeitos dos fármacos , Lenalidomida/farmacologia , Leucemia Mieloide Aguda , Síndromes Mielodisplásicas , Animais , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Ciclosporina/agonistas , Proteínas de Ligação a DNA/biossíntese , Regulação Leucêmica da Expressão Gênica/efeitos dos fármacos , Humanos , Fator de Transcrição Ikaros/biossíntese , Lenalidomida/agonistas , Leucemia Mieloide Aguda/tratamento farmacológico , Leucemia Mieloide Aguda/metabolismo , Leucemia Mieloide Aguda/patologia , Camundongos , Camundongos Endogâmicos NOD , Proteínas Musculares/biossíntese , Síndromes Mielodisplásicas/tratamento farmacológico , Síndromes Mielodisplásicas/metabolismo , Síndromes Mielodisplásicas/patologia , Proteínas de Neoplasias/biossíntese , Regulação para Cima/efeitos dos fármacos , Ensaios Antitumorais Modelo de Xenoenxerto
8.
Artigo em Inglês | MEDLINE | ID: mdl-32376613

RESUMO

Cyclophilins play a key role in the life cycle of coronaviruses. Alisporivir (Debio 025) is a nonimmunosuppressive analogue of cyclosporine with potent cyclophilin inhibition properties. Alisporivir reduced SARS-CoV-2 RNA production in a dose-dependent manner in Vero E6 cells, with a 50% effective concentration (EC50) of 0.46 ± 0.04 µM. Alisporivir inhibited a postentry step of the SARS-CoV-2 life cycle. These results justify rapidly conducting a proof-of-concept phase 2 trial with alisporivir in patients with SARS-CoV-2 infection.


Assuntos
Betacoronavirus/efeitos dos fármacos , Infecções por Coronavirus/tratamento farmacológico , Ciclofilinas/antagonistas & inibidores , Ciclosporina/farmacologia , Pneumonia Viral/tratamento farmacológico , Animais , Antivirais/farmacologia , Linhagem Celular , Chlorocebus aethiops , Humanos , Pandemias , Células Vero , Replicação Viral/efeitos dos fármacos
9.
Sheng Wu Gong Cheng Xue Bao ; 36(4): 605-611, 2020 Apr 25.
Artigo em Chinês | MEDLINE | ID: covidwho-151101

RESUMO

Cyclophilin A (CypA) is a widely distributed and highly conserved protein in organisms. It has peptidyl-prolyl cis/trans isomerase activity and is a receptor for cyclosporin A (CsA). Coronaviruses are enveloped, single-stranded, positive-sense RNA viruses. Seven types of coronaviruses are currently known to infect humans, among which SARS-CoV, MERS-CoV, and SARS-CoV-2 are fatal for humans. It is well established that CypA is essential for the replication of various coronaviruses such as SARS-CoV, CoV-229E, CoV-NL63, and FCoV. Additionally, CsA and its derivatives (ALV, NIM811, etc.) have obvious inhibitory effects on a variety of coronaviruses. These results suggest that CypA is a potential antiviral target and the existing drug CsA might be used as an anti-coronavirus drug. At the end of 2019, SARS-CoV-2 raged in China, which seriously theatern human health and causes huge economic lases. In view of this, we describe the effects of CypA on the replication of coronaviruses and the antiviral activities of its inhibitors, which will provide the scientific basis and ideas for the development of antiviral drugs for SARS-CoV-2.


Assuntos
Antivirais/farmacologia , Infecções por Coronavirus , Coronavirus/efeitos dos fármacos , Coronavirus/crescimento & desenvolvimento , Ciclofilina A/antagonistas & inibidores , Ciclosporina/farmacologia , Ciclosporina/uso terapêutico , Pandemias , Pneumonia Viral , Antivirais/uso terapêutico , Betacoronavirus/efeitos dos fármacos , Betacoronavirus/crescimento & desenvolvimento , Infecções por Coronavirus/tratamento farmacológico , Infecções por Coronavirus/epidemiologia , Infecções por Coronavirus/virologia , Ciclosporina/química , Humanos , Pneumonia Viral/tratamento farmacológico , Pneumonia Viral/epidemiologia , Pneumonia Viral/virologia , Vírus da SARS/efeitos dos fármacos , Vírus da SARS/crescimento & desenvolvimento , Replicação Viral/efeitos dos fármacos
10.
Sheng Wu Gong Cheng Xue Bao ; 36(4): 605-611, 2020 Apr 25.
Artigo em Chinês | MEDLINE | ID: mdl-32347055

RESUMO

Cyclophilin A (CypA) is a widely distributed and highly conserved protein in organisms. It has peptidyl-prolyl cis/trans isomerase activity and is a receptor for cyclosporin A (CsA). Coronaviruses are enveloped, single-stranded, positive-sense RNA viruses. Seven types of coronaviruses are currently known to infect humans, among which SARS-CoV, MERS-CoV, and SARS-CoV-2 are fatal for humans. It is well established that CypA is essential for the replication of various coronaviruses such as SARS-CoV, CoV-229E, CoV-NL63, and FCoV. Additionally, CsA and its derivatives (ALV, NIM811, etc.) have obvious inhibitory effects on a variety of coronaviruses. These results suggest that CypA is a potential antiviral target and the existing drug CsA might be used as an anti-coronavirus drug. At the end of 2019, SARS-CoV-2 raged in China, which seriously theatern human health and causes huge economic lases. In view of this, we describe the effects of CypA on the replication of coronaviruses and the antiviral activities of its inhibitors, which will provide the scientific basis and ideas for the development of antiviral drugs for SARS-CoV-2.


Assuntos
Antivirais/farmacologia , Infecções por Coronavirus , Coronavirus/efeitos dos fármacos , Coronavirus/crescimento & desenvolvimento , Ciclofilina A/antagonistas & inibidores , Ciclosporina/farmacologia , Ciclosporina/uso terapêutico , Pandemias , Pneumonia Viral , Antivirais/uso terapêutico , Betacoronavirus/efeitos dos fármacos , Betacoronavirus/crescimento & desenvolvimento , Infecções por Coronavirus/tratamento farmacológico , Infecções por Coronavirus/epidemiologia , Infecções por Coronavirus/virologia , Ciclosporina/química , Humanos , Pneumonia Viral/tratamento farmacológico , Pneumonia Viral/epidemiologia , Pneumonia Viral/virologia , Vírus da SARS/efeitos dos fármacos , Vírus da SARS/crescimento & desenvolvimento , Replicação Viral/efeitos dos fármacos
11.
Transplantation ; 104(9): e252-e259, 2020 09.
Artigo em Inglês | MEDLINE | ID: mdl-32217944

RESUMO

BACKGROUND: Despite the benefits of ex vivo lung perfusion (EVLP) such as lung reconditioning, preservation, and evaluation before transplantation, deleterious effects, including activation of proinflammatory cascades and alteration of metabolic profiles have been reported. Although patient outcomes have been favorable, further studies addressing optimal conditions are warranted. In this study, we investigated the role of the immunosuppressant drug cyclosporine A (CyA) in preserving mitochondrial function and subsequently preventing proinflammatory changes in lung grafts during EVLP. METHODS: Using rat heart-lung blocks after 1-hour cold preservation, an acellular normothermic EVLP system was established for 4 hours. CyA was added into perfusate at a final concentration of 1 µM. The evaluation included lung graft function, lung compliance, and pulmonary vascular resistance as well as biochemical marker measurement in the perfusate at multiple time points. After EVLP, single orthotopic lung transplantation was performed, and the grafts were assessed 2 hours after reperfusion. RESULTS: Lung grafts on EVLP with CyA exhibited significantly better functional and physiological parameters as compared with those without CyA treatment. CyA administration attenuated proinflammatory changes and prohibited glucose consumption during EVLP through mitigating mitochondrial dysfunction in lung grafts. CyA-preconditioned lungs showed better posttransplant lung early graft function and less inflammatory events compared with control. CONCLUSIONS: During EVLP, CyA administration can have a preconditioning effect through both its anti-inflammatory and mitochondrial protective properties, leading to improved lung graft preservation, which may result in enhanced graft quality after transplantation.


Assuntos
Ciclosporina/farmacologia , Imunossupressores/farmacologia , Transplante de Pulmão/métodos , Pulmão/irrigação sanguínea , Alarminas/antagonistas & inibidores , Animais , Cálcio/metabolismo , Nucleotídeos de Desoxiadenina , Masculino , Perfusão , Ratos , Ratos Endogâmicos Lew , Condicionamento Pré-Transplante
12.
Parasit Vectors ; 13(1): 94, 2020 Feb 21.
Artigo em Inglês | MEDLINE | ID: mdl-32085719

RESUMO

BACKGROUND: New therapeutic drugs are urgently needed against visceral leishmaniasis because current drugs, such as pentavalent antimonials and miltefosine, produce severe side effects and development of resistance. Whether cyclosporine A (CsA) and its derivatives can be used as therapeutic drugs for visceral leishmaniasis has been controversial for many years. METHODS: In this study, we evaluated the efficacy of CsA and its derivative, dihydrocyclosporin A (DHCsA-d), against promastigotes and intracellular amastigotes of Leishmania donovani. Sodium stibogluconate (SSG) was used as a positive control. RESULTS: Our results showed that DHCsA-d was able to inhibit the proliferation of L. donovani promastigotes (IC50: 21.24 µM and 12.14 µM at 24 h and 48 h, respectively) and intracellular amastigotes (IC50: 5.23 µM and 4.84 µM at 24 and 48 h, respectively) in vitro, but CsA treatment increased the number of amastigotes in host cells. Both DHCsA-d and CsA caused several alterations in the morphology and ultrastructure of L. donovani, especially in the mitochondria. However, DHCsA-d showed high cytotoxicity towards cells of the mouse macrophage cell line RAW264.7, with CC50 values of 7.98 µM (24 h) and 6.65 µM (48 h). Moreover, DHCsA-d could increase IL-12, TNF-α and IFN-γ production and decrease the levels of IL-10, IL-4, NO and H2O2 in infected macrophages. On the contrary, CsA decreased IL-12, TNF-α, and IFN-γ production and increased the levels of IL-10, IL-4, NO and H2O2 in infected macrophages. The expression of L. donovani cyclophilin A (LdCyPA) in promastigotes and intracellular amastigotes and the expression of cyclophilin A (CyPA) in RAW 264.7 cells were found to be significantly downregulated in the CsA-treated group compared to those in the untreated group. However, no significant changes in LdCyPA and CyPA levels were found after DHCsA-d or SSG treatment. CONCLUSIONS: Our findings initially resolved the dispute regarding the efficacy of CsA and DHCsA-d for visceral leishmaniasis treatment. CsA showed no significant inhibitory effect on intracellular amastigotes. DHCsA-d significantly inhibited promastigotes and intracellular amastigotes, but it was highly cytotoxic. Therefore, CsA and DHCsA-d are not recommended as antileishmanial drugs.


Assuntos
Antiprotozoários/farmacologia , Ciclosporina/farmacologia , Ciclosporinas/farmacologia , Leishmania donovani/efeitos dos fármacos , Leishmaniose Visceral/parasitologia , Animais , Avaliação Pré-Clínica de Medicamentos , Humanos , Interferon gama/imunologia , Interleucina-10/imunologia , Interleucina-2/imunologia , Leishmania donovani/crescimento & desenvolvimento , Leishmania donovani/fisiologia , Leishmaniose Visceral/tratamento farmacológico , Leishmaniose Visceral/imunologia , Macrófagos/imunologia , Macrófagos/parasitologia , Camundongos , Células RAW 264.7
13.
Braz Oral Res ; 34: e007, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32049108

RESUMO

The aim of this study was to assess the influence of cyclosporine administration on the repair of critical-sized calvaria defects (CSDs) in rat calvaria filled with diverse biomaterials. Sixty animals were divided into two groups: the control (CTR) group (saline solution) and the cyclosporine (CCP) group (cyclosporine, 10 mg/kg/day). These medications were administered daily by gavage, beginning 15 days before the surgical procedure and lasting until the day the animals were euthanized. A CSD (5 mm Ø) was made in the calvaria of each animal, which was allocated to one of 3 subgroups, according to the biomaterial used to fill the defect: coagulum (COA), deproteinized bovine bone (DBB), or biphasic calcium phosphate ceramics of hydroxyapatite and ß-phosphate tricalcium (HA/TCP). Euthanasia of the animals was performed 15 and 60 days after the surgical procedure (n = 5 animals/period/subgroup). Bone repair (formation) assessment was performed through microtomography and histometry, while the analyses of the expression of the BMP2, Osteocalcin, and TGFß1 proteins were performed using immunohistochemistry. The CSDs not filled with biomaterials demonstrated lower bone formation in the CCP group. At 15 days, less bone formation was observed in the CSDs filled with DBB, a smaller volume of mineralized tissue was observed in the CSDs filled with HA/TCP, and the expression levels of BMP2 and osteocalcin were lower in the CCP group compared to the CTR group. The use of cyclosporine impaired bone repair in CSD, and this effect can be partially explained by the suppression of BMP2 and osteocalcin expression.


Assuntos
Regeneração Óssea/efeitos dos fármacos , Substitutos Ósseos/farmacologia , Inibidores de Calcineurina/farmacologia , Ciclosporina/farmacologia , Osteogênese/efeitos dos fármacos , Animais , Proteína Morfogenética Óssea 2/análise , Imuno-Histoquímica , Masculino , Osteocalcina/análise , Distribuição Aleatória , Ratos , Reprodutibilidade dos Testes , Crânio/efeitos dos fármacos , Crânio/patologia , Fatores de Tempo , Fator de Crescimento Transformador beta1/análise , Microtomografia por Raio-X
14.
Crit Care Med ; 48(2): e123-e132, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-31939811

RESUMO

OBJECTIVES: Trauma predisposes to systemic sterile inflammation (systemic inflammatory response syndrome) as well as infection, but the mechanisms linking injury to infection are poorly understood. Mitochondrial debris contains formyl peptides. These bind formyl peptide receptor-1, trafficking neutrophils to wounds, initiating systemic inflammatory response syndrome, and wound healing. Bacterial formyl peptides, however, also attract neutrophils via formyl peptide receptor-1. Thus, mitochondrial formyl peptides might suppress neutrophils antimicrobial function. Also, formyl peptide receptor-1 blockade used to mitigate systemic inflammatory response syndrome might predispose to sepsis. We examined how mitochondrial formyl peptides impact neutrophils functions contributing to antimicrobial responses and how formyl peptide receptor-1 antagonists affect those functions. DESIGN: Prospective study of human and murine neutrophils and clinical cohort analysis. SETTING: University research laboratory and level 1 trauma center. PATIENTS: Trauma patients, volunteer controls. ANIMAL SUBJECTS: C57Bl/6, formyl peptide receptor-1, and formyl peptide receptor-2 knockout mice. INTERVENTIONS: Human and murine neutrophils functions were activated with autologous mitochondrial debris, mitochondrial formyl peptides, or bacterial formyl peptides followed by chemokines or leukotrienes. The experiments were repeated using formyl peptide receptor-1 antagonist cyclosporin H, "designer" human formyl peptide receptor-1 antagonists (POL7178 and POL7200), or anti-formyl peptide receptor-1 antibodies. Mouse injury/lung infection model was used to evaluate effect of formyl peptide receptor-1 inhibition. MEASUREMENTS AND MAIN RESULTS: Human neutrophils cytosolic calcium, chemotaxis, reactive oxygen species production, and phagocytosis were studied before and after exposure to mitochondrial debris, mitochondrial formyl peptides, and bacterial formyl peptides. Mitochondrial formyl peptide and bacterial formyl peptides had similar effects on neutrophils. Responses to chemokines and leukotrienes were suppressed by prior exposure to formyl peptides. POL7200 and POL7178 were specific antagonists of human formyl peptide receptor-1 and more effective than cyclosporin H or anti-formyl peptide receptor-1 antibodies. Formyl peptides inhibited mouse neutrophils responses to chemokines only if formyl peptide receptor-1 was present. Formyl peptide receptor-1 blockade did not inhibit neutrophils bacterial phagocytosis or reactive oxygen species production. Cyclosporin H increased bacterial clearance in lungs after injury. CONCLUSIONS: Formyl peptides both activate and desensitize neutrophils. Formyl peptide receptor-1 blockade prevents desensitization, potentially both diminishing systemic inflammatory response syndrome and protecting the host against secondary infection after tissue trauma or primary infection.


Assuntos
Proteínas Mitocondriais/imunologia , Ativação de Neutrófilo/imunologia , Receptores de Formil Peptídeo/antagonistas & inibidores , Animais , Ciclosporina/farmacologia , Humanos , Lesão Pulmonar/fisiopatologia , Camundongos , Camundongos Endogâmicos C57BL , Neutrófilos/imunologia , Infecções Respiratórias/fisiopatologia
15.
Clin Sci (Lond) ; 134(2): 239-259, 2020 Jan 31.
Artigo em Inglês | MEDLINE | ID: mdl-31943002

RESUMO

Mitochondrial stress has been widely observed in diabetic kidney disease (DKD). Cyclophilin D (CypD) is a functional component of the mitochondrial permeability transition pore (mPTP) which allows the exchange of ions and solutes between the mitochondrial matrix to induce mitochondrial swelling and activation of cell death pathways. CypD has been successfully targeted in other disease contexts to improve mitochondrial function and reduced pathology. Two approaches were used to elucidate the role of CypD and the mPTP in DKD. Firstly, mice with a deletion of the gene encoding CypD (Ppif-/-) were rendered diabetic with streptozotocin (STZ) and followed for 24 weeks. Secondly, Alisporivir, a CypD inhibitor was administered to the db/db mouse model (5 mg/kg/day oral gavage for 16 weeks). Ppif-/- mice were not protected against diabetes-induced albuminuria and had greater glomerulosclerosis than their WT diabetic littermates. Renal hyperfiltration was lower in diabetic Ppif-/- as compared with WT mice. Similarly, Alisporivir did not improve renal function nor pathology in db/db mice as assessed by no change in albuminuria, KIM-1 excretion and glomerulosclerosis. Db/db mice exhibited changes in mitochondrial function, including elevated respiratory control ratio (RCR), reduced mitochondrial H2O2 generation and increased proximal tubular mitochondrial volume, but these were unaffected by Alisporivir treatment. Taken together, these studies indicate that CypD has a complex role in DKD and direct targeting of this component of the mPTP will likely not improve renal outcomes.


Assuntos
Ciclofilina D/metabolismo , Diabetes Mellitus Experimental/metabolismo , Nefropatias/metabolismo , Proteínas de Transporte da Membrana Mitocondrial/metabolismo , Albuminúria/genética , Albuminúria/metabolismo , Animais , Ciclofilina D/antagonistas & inibidores , Ciclofilina D/genética , Ciclosporina/farmacologia , Diabetes Mellitus Experimental/genética , Nefropatias Diabéticas/genética , Nefropatias Diabéticas/metabolismo , Peróxido de Hidrogênio/metabolismo , Rim/efeitos dos fármacos , Rim/metabolismo , Rim/patologia , Nefropatias/genética , Camundongos Endogâmicos C57BL , Camundongos Knockout , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Proteínas de Transporte da Membrana Mitocondrial/genética , ATPases Translocadoras de Prótons/metabolismo
16.
Clin Nucl Med ; 45(2): e69-e71, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31833924

RESUMO

A 30-year-old woman presented with persistent fever and elevated lactate dehydrogenase levels without skin rash. F-FDG PET/CT revealed FDG uptake in subcutaneous tissues in the forearm, buttocks, and lower limbs, whereas the trunk was unaffected. She was diagnosed with subcutaneous panniculitis-like T-cell lymphoma based on a PET/CT-guided subcutaneous tissue biopsy from the thigh. Although conventional cytotoxic agent-based chemotherapies failed to achieve disease remission, subsequent cyclosporine A treatment promptly resolved the fever and laboratory abnormalities. Remarkably, abnormal FDG uptake disappeared entirely on follow-up PET/CT, demonstrating its utility in the evaluations of responses to emerging cyclosporine A-based treatments in subcutaneous panniculitis-like T-cell lymphoma.


Assuntos
Ciclosporina/farmacologia , Fluordesoxiglucose F18 , Linfoma de Células T/diagnóstico por imagem , Linfoma de Células T/tratamento farmacológico , Paniculite/diagnóstico por imagem , Paniculite/tratamento farmacológico , Tomografia Computadorizada com Tomografia por Emissão de Pósitrons , Adulto , Ciclosporina/uso terapêutico , Feminino , Humanos , Linfoma de Células T/patologia , Paniculite/patologia , Resultado do Tratamento
17.
J Stroke Cerebrovasc Dis ; 29(2): 104562, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31836361

RESUMO

INTRODUCTION: Recent studies have indicated that the damaging effects of stroke are not only limited to the brain. We sought to examine the changes of liver and renal enzymes in the acute phase of ischemic stroke and to investigate possible explanations and therapeutic options, concerning in particular the functional alterations of peripheral organs after administration of an anti-inflammatory agent. MATERIAL/METHODS: Twelve-week-old Wistar male rats were randomly divided into control and Cyclosporine groups (n = 10 each). Cyclosporine was given orally by gavage for 5 days prior to cerebral ischemia at a total volume of 15 mg/kg/day. All animals were subjected to 60 minutes focal ischemia by filament occlusion of the middle cerebral artery. Serum concentrations of Creatinine, Urea, SGOT, SGPT, and γGT were determined at the time before surgery and after 60 minutes brain ischemia. RESULTS: Comparing data of 2 time-points, in both groups the serum liver enzyme levels increased progressively during the ischemic period. The liver enzymes and Urea were significantly lower in the Cyclosporine group than in the control group and the levels of Creatinine were slightly higher in the Cyclosporine group, in both time-points. CONCLUSIONS: The detection of high liver enzyme serum levels in the acute phase of ischemic stroke implies the secondary effect of cerebral infraction on the peripheral organs and particularly on the liver function. Cyclosporine seems to exhibit a protective activity and to affect both liver and renal function after ischemic stroke.


Assuntos
Anti-Inflamatórios/farmacologia , Isquemia Encefálica/tratamento farmacológico , Ciclosporina/farmacologia , Nefropatias/prevenção & controle , Rim/efeitos dos fármacos , Hepatopatias/prevenção & controle , Fígado/efeitos dos fármacos , Acidente Vascular Cerebral/tratamento farmacológico , Animais , Biomarcadores/sangue , Isquemia Encefálica/sangue , Isquemia Encefálica/complicações , Isquemia Encefálica/fisiopatologia , Modelos Animais de Doenças , Rim/enzimologia , Rim/fisiopatologia , Nefropatias/sangue , Nefropatias/etiologia , Nefropatias/fisiopatologia , Fígado/enzimologia , Fígado/fisiopatologia , Hepatopatias/sangue , Hepatopatias/etiologia , Hepatopatias/fisiopatologia , Masculino , Ratos Wistar , Acidente Vascular Cerebral/sangue , Acidente Vascular Cerebral/complicações , Acidente Vascular Cerebral/fisiopatologia , Fatores de Tempo
18.
Xenobiotica ; 50(5): 614-619, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-31573401

RESUMO

1. Voriconazole is known to display highly variable pharmacokinetics affecting treatment efficacy and safety. This study aimed to identify the factors causing the variation of voriconazole concentration in patients with allogeneic hematopoietic stem cell transplantation.2. The data of patients was collected, including clinical characteristics and voriconazole concentrations. A total of 5 single nucleotide polymorphisms of 3 candidate genes (CYP2C19, ABCC2, ABCG2) related to voriconazole metabolism were genotyped by MassArray method. The correlation between polymorphisms and voriconazole concentration was analyzed.3. A total of 244 voriconazole concentrations of 43 patients were included in this study. The voriconazole concentration was significantly correlated with patients' total bile acid (p = 0.001) and cyclosporin A (p < 0.001). The median concentration of the CYP2C19 normal metabolizers was remarkably lower than poor metabolizers (0.86 vs 2.27 µg/mL). The median concentration of ABCC2 rs2273697 GG genotype carriers was significantly higher than that of GA genotype carriers (p = 0.026).4. The variability of voriconazole concentration is partially explained by total bile acid, metabolic types of CYP2C19. The voriconazole concentration of CYP2C19 normal metabolizers is likely to be lower than 1.0 µg/mL and thus at risk of infection due to inadequate treatment.


Assuntos
Antifúngicos/farmacologia , Ciclosporina/farmacologia , Citocromo P-450 CYP2C19/genética , Voriconazol/metabolismo , Adulto , Antifúngicos/metabolismo , Ciclosporina/metabolismo , Transplante de Células-Tronco Hematopoéticas , Humanos , Proteínas Associadas à Resistência a Múltiplos Medicamentos , Variantes Farmacogenômicos , Polimorfismo de Nucleotídeo Único
19.
Biomed Pharmacother ; 123: 109747, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31881484

RESUMO

Psoriasis is a T cells mediated chronic skin inflammation in which helper T (Th) cells play in critical roles in its pathogenesis. Taxifolin (TXL) has been discovered to exert various pharmacological activities. In this study, we wished to observe whether TXL had potential activities on psoriasis, and how it works. We found that TXL can inhibit LPS-induced abnormal proliferation in Hacat cell line, ant also significantly alleviate the IMQ-induced psoriasis in BALB/c mice, comparing with the control group. Although TXL has no significant effects on the ratio of total T cells in skin draining lymph nodes (SDLN), it decreases the ratio of pro-inflammatory Th1 and Th17 cells, both in skin lesions and SDLN. Our results also disclosed that TXL may regulate Th cells differentiation by inhibiting the transcript factors, including T-bet, GATA-3 and RORγt. Further data show that TXL can inhibit Notch1 and Jak2/Stat3 signal pathways. In summary, TXL may be able to treat psoriasis by regulating Th cells differentiation via inhibiting Notch1 and Jak2/Stat3 pathways.


Assuntos
Dermatite/tratamento farmacológico , Imiquimode/farmacologia , Janus Quinase 2/metabolismo , Psoríase/tratamento farmacológico , Quercetina/análogos & derivados , Receptor Notch1/metabolismo , Fator de Transcrição STAT3/metabolismo , Animais , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Ciclosporina/farmacologia , Citocinas/metabolismo , Humanos , Inflamação , Queratinócitos , Lipopolissacarídeos/farmacologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Modelos Animais , Psoríase/induzido quimicamente , Quercetina/química , Quercetina/farmacologia , Transdução de Sinais/efeitos dos fármacos , Pele/efeitos dos fármacos , Linfócitos T/efeitos dos fármacos , Linfócitos T/metabolismo , Fatores de Transcrição/metabolismo
20.
ACS Appl Mater Interfaces ; 12(3): 3438-3444, 2020 Jan 22.
Artigo em Inglês | MEDLINE | ID: mdl-31877011

RESUMO

In recent years, rapid development of induced pluripotent stem cell (iPSC) technology has provided good technical support for the study of human cardiovascular disease (CVD). In this work, a mimetic cell membrane and drug carrier OPFL system containing photoactive oligo(p-phenylene vinylene) functionalized with phospholipid units (OPV-lipid) was prepared for functional regulation of iPSC-derived cardiomyocytes. OPFL bound to the cell membrane of iPSC-derived human cardiomyocytes and significantly enhanced delivery of cyclosporin A (CsA) into these cells, which promoted the regulation of mitochondrial calcium levels and membrane potential by CsA. This led to the protection of the mitochondrial structure and function, thus reducing apoptosis of iPSC-derived cardiomyocytes and achieving the effect of treating CVD. OPFL not only acts as a fluorescent probe for cell imaging and visualization of the drug delivery process but also provides a tool to deliver lipid-insoluble drugs throughout the cell membrane. Benefiting from good biocompatibility, facile operation, and a visible and controllable cell uptake process, OPFL has good potential to be a powerful tool in future basic and clinical research applications involving iPSCs.


Assuntos
Células-Tronco Pluripotentes Induzidas/citologia , Polivinil/química , Substâncias Protetoras/química , Apoptose/efeitos dos fármacos , Cálcio/metabolismo , Membrana Celular/metabolismo , Células Cultivadas , Ciclosporina/farmacologia , Sistemas de Liberação de Medicamentos , Humanos , Células-Tronco Pluripotentes Induzidas/efeitos dos fármacos , Células-Tronco Pluripotentes Induzidas/metabolismo , Potenciais da Membrana , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Miócitos Cardíacos/citologia , Miócitos Cardíacos/efeitos dos fármacos , Miócitos Cardíacos/metabolismo , Fosfolipídeos/química , Polivinil/metabolismo
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