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1.
Pain ; 163(2): e202-e214, 2022 02 01.
Artigo em Inglês | MEDLINE | ID: mdl-34252912

RESUMO

ABSTRACT: Gain-of-function and loss-of-function mutations in Nav1.7 cause chronic pain and pain insensitivity, respectively. The preferential expression of Nav1.7 in the peripheral nervous system and its role in human pain signaling make Nav1.7 a promising target for next-generation pain therapeutics. However, pharmacological agents have not fully recapitulated these pain phenotypes, and because of the lack of subtype-selective molecular modulators, the role of Nav1.7 in the perception of pain remains poorly understood. Scorpion venom is an excellent source of bioactive peptides that modulate various ion channels, including voltage-gated sodium (Nav) channels. Here, we demonstrate that Buthus martensii Karsch scorpion venom (BV) elicits pain responses in mice through direct enhancement of Nav1.7 activity and have identified Makatoxin-3, an α-like toxin, as a critical component for BV-mediated effects on Nav1.7. Blocking other Nav subtypes did not eliminate BV-evoked pain responses, supporting the pivotal role of Nav1.7 in BV-induced pain. Makatoxin-3 acts on the S3-S4 loop of voltage sensor domain IV (VSD4) of Nav1.7, which causes a hyperpolarizing shift in the steady-state fast inactivation and impairs inactivation kinetics. We also determined the key residues and structure-function relationships for the toxin-channel interactions, which are distinct from those of other well-studied α toxins. This study not only reveals a new mechanism underlying BV-evoked pain but also enriches our knowledge of key structural elements of scorpion toxins that are pivotal for toxin-Nav1.7 interactions, which facilitates the design of novel Nav1.7 selective modulators.


Assuntos
Dor Crônica , Picadas de Escorpião , Venenos de Escorpião , Animais , Dor Crônica/genética , Humanos , Camundongos , Fenótipo , Venenos de Escorpião/química , Venenos de Escorpião/toxicidade , Escorpiões
2.
Exp Ther Med ; 20(3): 2845-2855, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32765781

RESUMO

The use of Shi Xiao San (SXS), composed of Pollen Typhae Angustifoliae and Faeces Trogopterori, can be traced back to the Song dynasty. Traditionally, SXS has been used to treat irregular menstruation, pelvic pain, progressive dysmenorrhea, and postpartum lochiorrhea. The management of adenomyosis (AM) is challenging and to the best of our knowledge there are currently no effective therapeutic strategies. Therefore, the aim of the present study was to investigate the effect of SXS on the development of adenomyosis in a mouse model. AM was induced in 60 neonatal female ICR mice by administering tamoxifen; 10 randomly selected mice were used for model identification via histopathological examination and 10 mice treated with the solvent alone were used as the normal controls. A total of sixty days after birth, the mice treated with AM were randomly divided into four groups and administered one of the following treatments: Low-dose SXS (55 mg/kg); high-dose SXS (110 mg/kg); danazol (1 mg/20 g body weight); or no treatment (model group); at the same time, the normal control group received no treatment. After 2 months of treatment, hotplate and tail-flick tests were used to assess the response to noxious thermal stimuli in the mice, and plasma samples were collected to measure corticosterone levels. Hematoxylin and eosin staining scores of myometrial infiltration and the number of AM nodules were evaluated. Furthermore, the expression of genes associated with AM-related pain was also analyzed. The results from the present study indicated that treatment with SXS decreased myometrial infiltration, alleviated generalized hyperalgesia, and lowered plasma corticosterone levels in mice with induced AM. These findings suggest that SXS effectively attenuated the development of AM, and may serve as a promising treatment approach for AM treatment.

3.
J Immunother Cancer ; 7(1): 326, 2019 11 27.
Artigo em Inglês | MEDLINE | ID: mdl-31775862

RESUMO

BACKGROUND: It is unclear whether plant-derived extracellular vesicles (EVs) can mediate interspecies communication with mammalian cells. Tumor-associated macrophages (TAMs) display a continuum of different polarization states between tumoricidal M1 phenotype and tumor-supportive M2 phenotypes, with a lower M1/M2 ratio correlating with tumor growth, angiogenesis and invasion. We investigated whether EVs from ginseng can alter M2-like polarization both in vitro and in vivo to promote cancer immunotherapy. METHODS: A novel EVs-liked ginseng-derived nanoparticles (GDNPs) were isolated and characterized from Panax ginseng C. A. Mey. Using GDNPs as an immunopotentiator for altering M2 polarized macrophages, we analyzed associated surface markers, genes and cytokines of macrophages treated with GDNPs. Mice bearing B16F10 melanoma were treated with GDNPs therapy. Tumor growth were assessed, and TAM populations were evaluated by FACS and IF. RESULTS: GDNPs significantly promoted the polarization of M2 to M1 phenotype and produce total reactive oxygen species, resulting in increasing apoptosis of mouse melanoma cells. GDNP-induced M1 polarization was found to depend upon Toll-like receptor (TLR)-4 and myeloid differentiation antigen 88 (MyD88)-mediated signaling. Moreover, ceramide lipids and proteins of GDNPs may play an important role in macrophage polarization via TLR4 activation. We found that GDNPs treatment significantly suppressed melanoma growth in tumor-bearing mice with increased presence of M1 macrophages detected in the tumor tissue. CONCLUSIONS: GDNPs can alter M2 polarization both in vitro and in vivo, which contributes to an antitumor response. The polarization of macrophages induced by GDNPs is largely dependent on TLR4 and MyD88 signalling. GDNPs as an immunomodulator participate in mammalian immune response and may represent a new class of nano-drugs in cancer immunotherapy.


Assuntos
Antineoplásicos Fitogênicos/farmacologia , Ativação de Macrófagos/efeitos dos fármacos , Ativação de Macrófagos/imunologia , Macrófagos/efeitos dos fármacos , Macrófagos/imunologia , Nanopartículas , Panax/química , Extratos Vegetais/farmacologia , Animais , Antineoplásicos Fitogênicos/química , Antineoplásicos Fitogênicos/isolamento & purificação , Linhagem Celular Tumoral , Centrifugação com Gradiente de Concentração , Modelos Animais de Doenças , Estabilidade de Medicamentos , Humanos , Lipídeos/química , Macrófagos/metabolismo , Masculino , Melanoma/tratamento farmacológico , Melanoma/imunologia , Melanoma/metabolismo , Melanoma/patologia , Melanoma Experimental , Camundongos , Fator 88 de Diferenciação Mieloide/metabolismo , Nanopartículas/química , Fagocitose , Extratos Vegetais/química , Extratos Vegetais/isolamento & purificação , Distribuição Tecidual , Receptor 4 Toll-Like/metabolismo , Ensaios Antitumorais Modelo de Xenoenxerto
4.
Free Radic Biol Med ; 120: 13-24, 2018 05 20.
Artigo em Inglês | MEDLINE | ID: mdl-29530794

RESUMO

Oxaliplatin-induced peripheral neuropathy (OIPN) is a severe, dose-limiting toxicity associated with cancer chemotherapy. The efficacy of antioxidant administration in OIPN is debatable, as the promising preliminary results obtained with a number of antioxidants have not been confirmed in larger clinical trials. Besides its antioxidant activity, the transcription factor, nuclear factor-erythroid 2 (NF-E2) p45-related factor 2 (Nrf2) plays a crucial role in the maintenance of mitochondrial homeostasis, and mitochondrial dysfunction is a key contributor to OIPN. Here, we have investigated the protective properties of Nrf2 in OIPN. Nrf2-/- mice displayed severe mechanical allodynia and cold sensitivity and thus experienced increased peripheral nervous system injury compared to Nrf2+/+ mice. Furthermore, Nrf2 knockout aggravated oxaliplatin-induced reactive oxygen species production, decreased the mitochondrial membrane potential, led to abnormal intracellular calcium levels, and induced cytochrome c-related apoptosis and overexpression of the TRP protein family. Sulforaphane-induced activation of the Nrf2 signaling pathway alleviated morphological alterations, mitochondrial dysfunction in dorsal root ganglion neurons, and nociceptive sensations in mice. Our findings reveal that Nrf2 may play a critical role in ameliorating OIPN, through protection of mitochondrial function by alleviating oxidative stress and inhibiting TRP protein family expression. This suggests that pharmacological or therapeutic activation of Nrf2 may be used to prevent or slow down the progression of OIPN.


Assuntos
Antineoplásicos/toxicidade , Fator 2 Relacionado a NF-E2/metabolismo , Oxaliplatina/toxicidade , Doenças do Sistema Nervoso Periférico/induzido quimicamente , Doenças do Sistema Nervoso Periférico/metabolismo , Animais , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Mitocôndrias/metabolismo , Estresse Oxidativo/fisiologia
5.
Mol Pain ; 14: 1744806918761238, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29424271

RESUMO

Background Several studies have shown that scorpion venom peptide BmK AGAP has an analgesic activity. Our previous study also demonstrated that intraplantar injection of BmK AGAP ameliorates formalin-induced spontaneous nociceptive behavior. However, the effect of intrathecal injection of BmK AGAP on nociceptive processing is poorly understood. Methods We investigated the effects of intrathecal injection of BmK AGAP on spinal nociceptive processing induced by chronic constrictive injury or formalin. Thermal hyperalgesia and mechanical allodynia were measured using radiant heat and the von Frey filaments test. Formalin-induced spontaneous nociceptive behavior was also investigated. C-Fos expression was assessed by immunohistochemistry. Phosphorylated mitogen-activated protein kinase (p-MAPK) expression was monitored by Western blot assay. Results Intrathecal injection of BmK AGAP reduced chronic constrictive injury-induced neuropathic pain behavior and pain from formalin-induced inflammation, accompanied by decreased expression of spinal p-MAPKs and c-Fos protein. The results of combining low doses of different MAPK inhibitor (U0126, SP600125, or SB203580; 0.1 µg for each inhibitor) with a low dose of BmK AGAP (0.2 µg) suggested that BmK AGAP could potentiate the effects of MAPK inhibitors on inflammation-associated pain. Conclusion Our results demonstrate that intrathecal injection of BmK AGAP produces a sensory-specific analgesic effect via a p-MAPK-dependent mechanism.


Assuntos
Analgésicos/uso terapêutico , Proteínas Quinases Ativadas por Mitógeno/antagonistas & inibidores , Venenos de Escorpião/uso terapêutico , Sensação , Medula Espinal/enzimologia , Analgésicos/farmacologia , Animais , Constrição , Modelos Animais de Doenças , Regulação para Baixo/efeitos dos fármacos , Formaldeído , Hiperalgesia/complicações , Hiperalgesia/tratamento farmacológico , Hiperalgesia/patologia , Inflamação/complicações , Inflamação/tratamento farmacológico , Inflamação/patologia , Injeções Espinhais , Masculino , Camundongos , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Neuralgia/complicações , Neuralgia/tratamento farmacológico , Neuralgia/patologia , Fosforilação/efeitos dos fármacos , Inibidores de Proteínas Quinases/farmacologia , Inibidores de Proteínas Quinases/uso terapêutico , Proteínas Proto-Oncogênicas c-fos/metabolismo , Venenos de Escorpião/administração & dosagem , Venenos de Escorpião/farmacologia , Sensação/efeitos dos fármacos , Medula Espinal/efeitos dos fármacos , Medula Espinal/patologia
6.
J Ethnopharmacol ; 213: 311-320, 2018 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-29180043

RESUMO

ETHNOPHARMACOLOGICAL RELEVANCE: Bungarus multicinctus snake belongs to Elapidae family and is widely distributed in southern China. It is widely used in traditional Chinese medicine with the effect of dispelling wind and removing obstruction in the meridians. Moreover, it is also as a chief ingredient of many polyherbal formulations for the treatment of cancer. AIM OF THE STUDY: To evaluate the antitumor activity of Bungarus multicinctus snake venom components and isolate, characterize the most effective anti-tumor component of Bungarus multicinctus snake venom. MATERIALS AND METHODS: The in vitro antitumor activity of Bungarus multicinctus venom components was detected by cytotoxicity assay and cell apoptosis assay. A unique LAAO from Bungarus multicinctus venom named as BM-Apotxin was isolated and characterized by Sephadex G-75 gel filtration, Sephadex G-25 desalting, Q ion-exchange chromatography and subsequent amino acids sequence determination. The LAAO activity and enzyme kinetics of BM-Apotxin was detected by microplate assay. RESULTS: BM-Apotxin, a 65KDa glycoprotein, which contributed to the most anti-tumor effects of Bungarus multicinctus venom. BM-Apotxin can selectively kill tumor cells, with less cytotoxicity to the normal cells. BM-Apotxin is an L-amino acid oxidase (LAAO) with high sequence identity to other snake venom LAAOs. Its anti-tumor activity is mainly due to the hydrogen peroxide produced from LAAO oxidation. But the catalase did not reverse its anti-tumor effect completely. Like other snake venom LAAOs, BM-Apotxin can oxidize many L amino acids, not D amino acids. The optimum substrate for BM-Apotxin is L-Phe. Moreover, BM-Apotxin deglycosylation can significantly reduce the LAAO activity and anti-tumor activity of BM-Apotxin. CONCLUSION: This study will facilitate the study on anti-tumor mechanism of snake venom and drug development based on Bungarus multicinctus venom.


Assuntos
Bungarus , Venenos Elapídicos/farmacologia , L-Aminoácido Oxidase/isolamento & purificação , L-Aminoácido Oxidase/farmacologia , Sequência de Aminoácidos , Animais , Antineoplásicos/isolamento & purificação , Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Linhagem Celular , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Venenos Elapídicos/química , Humanos , L-Aminoácido Oxidase/química
7.
Cancer Res ; 77(4): 926-936, 2017 02 15.
Artigo em Inglês | MEDLINE | ID: mdl-28011619

RESUMO

Aberrant expression of thioredoxin 1 (Trx1) plays an important role in cancer initiation and progression and has gained attention as an anticancer drug target. Here we report that the recently discovered natural diterpenoid isoforretin A (IsoA) significantly inhibits Trx1 activity and mediates anticancer effects in multiple preclinical settings. The inhibitory effect of IsoA was antagonized by free radical scavengers polyethylene glycol-catalase, polyethylene glycol superoxide dismutase, thiol-based antioxidants N-acetylcysteine and glutathione. Mass spectrometry analysis revealed that the mechanism of action was based on direct conjugation of IsoA to the Cys32/Cys35 residues of Trx1. This conjugation event attenuated reversible thiol reduction of Trx1, leading to ROS accumulation and a broader degradation of thiol redox homeostasis in cancer cells. Extending these in vitro findings, we documented that IsoA administration inhibited the growth of HepG2 tumors in a murine xenograft model of hepatocellular carcinoma. Taken together, our findings highlight IsoA as a potent bioactive inhibitor of Trx1 and a candidate anticancer natural product. Cancer Res; 77(4); 926-36. ©2016 AACR.


Assuntos
Antineoplásicos/farmacologia , Diterpenos/farmacologia , Espécies Reativas de Oxigênio/metabolismo , Tiorredoxinas/antagonistas & inibidores , Animais , Apoptose/efeitos dos fármacos , Quebras de DNA de Cadeia Dupla , Diterpenos/uso terapêutico , Células Hep G2 , Humanos , MAP Quinase Quinase Quinase 5/fisiologia , Sistema de Sinalização das MAP Quinases , Camundongos , Camundongos Endogâmicos BALB C , Neoplasias Experimentais/tratamento farmacológico , Neoplasias Experimentais/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Polietilenoglicóis/farmacologia , Superóxido Dismutase/farmacologia , Ensaios Antitumorais Modelo de Xenoenxerto
8.
Sci Rep ; 6: 35030, 2016 10 13.
Artigo em Inglês | MEDLINE | ID: mdl-27733755

RESUMO

As one of the three major human pathogens that cause schistosomiasis, Schistosoma japonicum is the only one that is endemic in China. Despite great progress on schistosomiasis control over the past 50 years in China, S. japonicum transmission still occurs in certain endemic regions, which causes significant public health problems and enormous economic losses. During different life stages, parasites are able to survive dramatic osmolality changes between its vector, fresh water, and mammal host. However, the molecular mechanism of parasite osmoregulation remains unknown. To address this challenging question, we report the first cloning of an S. japonicum aquaglyceroporin (SjAQP) from an isolate from Jiangsu province, China. Expressing SjAQP in Xenopus oocytes facilitated the permeation of water, glycerol, and urea. The water permeability of SjAQP was inhibited by 1 mM HgCl2, 3 mM tetraethylammonium, 1 mM ZnCl2, and 1 mM CuSO4. SjAQP was constitutively expressed throughout the S. japonicum life cycle, including in the egg, miracidia, cercaria, and adult stages. The highest expression was detected during the infective cercaria stage. Our results suggest that SjAQP plays a role in osmoregulation throughout the S. japonicum life cycle, especially during cercariae transformation, which enables parasites to survive osmotic challenges.


Assuntos
Aquagliceroporinas/genética , Aquagliceroporinas/metabolismo , Clonagem Molecular/métodos , Schistosoma japonicum/fisiologia , Animais , Animais Geneticamente Modificados , Água Doce/parasitologia , Regulação da Expressão Gênica no Desenvolvimento , Proteínas de Helminto/genética , Proteínas de Helminto/metabolismo , Humanos , Camundongos , Oócitos/metabolismo , Osmorregulação , Schistosoma japonicum/metabolismo , Xenopus/genética
9.
Sci Rep ; 6: 32496, 2016 08 31.
Artigo em Inglês | MEDLINE | ID: mdl-27578261

RESUMO

Observation of tongue coating, a foundation for clinical diagnosis and treatment in traditional Chinese medicine (TCM), is a major indicator of the occurrence, development, and prognosis of disease. The biological basis of tongue diagnosis and relationship between the types and microorganisms of tongue coating remain elusive. Thirteen chronic erosive gastritis (CEG) patients with typical yellow tongue coating (YTC) and ten healthy volunteers with thin white tongue coating (WTC) were included in this study. Patients were provided a 2-course targeted treatment of a herbal medicine Ban Xia Xie Xin decoction, traditionally prescribed for CEG patients with YTC, to evaluate the relationship between tongue coating microbiota and diagnosis of CEG with typical YTC. The tongue coating segregation structure was determined using Illumina Miseq sequencing of the V4-V5 region of the 16S ribosomal RNA gene. Bacillus was significantly observed only in CEG patients with YTC, but not in patients who received the decoction. YTC (n = 22) and WTC (n = 29) samples were collected for bacterial culturing to illustrate the relationship between Bacillus and YTC. The Bacillus positivity rate of YTC samples was 72.7%; Bacillus was not observed in WTC samples. In conclusion, Bacillus was strongly associated with YTC.


Assuntos
Bacillus/isolamento & purificação , Medicamentos de Ervas Chinesas/uso terapêutico , Gastrite/diagnóstico , Gastrite/tratamento farmacológico , Fitoterapia/métodos , Extratos Vegetais/uso terapêutico , Bacillus/classificação , Bacillus/genética , Bactérias/classificação , Bactérias/genética , Bactérias/isolamento & purificação , Estudos de Casos e Controles , Doença Crônica , Cor , Gastrite/microbiologia , Gastrite/patologia , Humanos , Medicina Tradicional Chinesa , Microbiota/genética , Análise de Componente Principal , RNA Ribossômico 16S/genética , Língua/microbiologia , Língua/patologia
10.
Sci Rep ; 6: 32206, 2016 08 24.
Artigo em Inglês | MEDLINE | ID: mdl-27553905

RESUMO

Sulforaphane, a naturally occurring compound found in cruciferous vegetables, has been shown to be neuroprotective in several neurological disorders. In this study, we sought to investigate the potential protective effects and associated molecular mechanisms of sulforaphane in an in vivo Parkinson's disease (PD) model, based on rotenone-mediated neurotoxicity. Our results showed that sulforaphane inhibited rotenone-induced locomotor activity deficiency and dopaminergic neuronal loss. Additionally, sulforaphane treatment inhibited the rotenone-induced reactive oxygen species production, malondialdehyde (MDA) accumulation, and resulted in an increased level of total glutathione and reduced glutathione (GSH): oxidized glutathione (GSSG) in the brain. Western blot analysis illustrated that sulforaphane increased the expression of nuclear factor (erythroid-derived 2)-like 2 (Nrf2), heme oxygenase-1 (HO-1), and NAD(P)H quinone oxidoreductase (NQO1), the latter two of which are anti-oxidative enzymes. Moreover, sulforaphane treatment significantly attenuated rotenone-inhibited mTOR-mediated p70S6K and 4E-BP1 signalling pathway, as well as neuronal apoptosis. In addition, sulforaphane rescued rotenone-inhibited autophagy, as detected by LC3-II. Collectively, these findings demonstrated that sulforaphane exert neuroprotective effect involving Nrf2-dependent reductions in oxidative stress, mTOR-dependent inhibition of neuronal apoptosis, and the restoration of normal autophagy. Sulforaphane appears to be a promising compound with neuroprotective properties that may play an important role in preventing PD.


Assuntos
Autofagia/efeitos dos fármacos , Isotiocianatos/farmacologia , Fármacos Neuroprotetores/farmacologia , Síndromes Neurotóxicas/prevenção & controle , Rotenona/toxicidade , Animais , Linhagem Celular , Modelos Animais de Doenças , Neurônios Dopaminérgicos/efeitos dos fármacos , Neurônios Dopaminérgicos/patologia , Glutationa/metabolismo , Humanos , Masculino , Camundongos Endogâmicos BALB C , Fator 2 Relacionado a NF-E2/metabolismo , Síndromes Neurotóxicas/tratamento farmacológico , Síndromes Neurotóxicas/metabolismo , Doença de Parkinson/etiologia , Sulfóxidos , Serina-Treonina Quinases TOR/metabolismo
11.
Biotechnol Lett ; 37(12): 2461-6, 2015 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-26303431

RESUMO

OBJECTIVES: To identify an efficient in vitro refolding method to generate highly active His6-tagged scorpion toxin antitumor-analgesic peptide (AGAP) isolated from Escherichia coli inclusion bodies. RESULTS: N- and C-Terminal His6-tagged recombinant (r) AGAP (NHis6-rAGAP and CHis6-rAGAP, respectively) were expressed in E. coli; the purification and refolding conditions were optimized. CHis6-rAGAP, but not NHis6-rAGAP, exhibited significant in vitro antihepatoma activity that was much greater than that of rAGAP produced using SUMO fusion technology (IC50, 0.4 ± 0.08 vs. 1.8 ± 0.3 µM). CHis6-rAGAP also showed significant inhibition of tumor growth in a mouse xenograft model of human hepatoma and inhibition of neuronal excitability, demonstrated by blockage of voltage-sensitive tetrodotoxin-resistant (TTX-R) sodium currents in acute isolated dorsal root ganglion neurons. CONCLUSIONS: This refolding protocol optimized for C-terminal His6-tagged scorpion rAGAP is potentially applicable to similar long-chain and cysteine-rich toxins.


Assuntos
Histidina/metabolismo , Dobramento de Proteína , Proteínas Recombinantes de Fusão/metabolismo , Venenos de Escorpião/metabolismo , Animais , Carcinoma Hepatocelular/tratamento farmacológico , Modelos Animais de Doenças , Escherichia coli/genética , Expressão Gênica , Xenoenxertos , Histidina/genética , Histidina/isolamento & purificação , Histidina/uso terapêutico , Humanos , Camundongos , Neurônios/efeitos dos fármacos , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/isolamento & purificação , Proteínas Recombinantes de Fusão/uso terapêutico , Venenos de Escorpião/genética , Venenos de Escorpião/isolamento & purificação , Venenos de Escorpião/uso terapêutico , Resultado do Tratamento , Canais de Sódio Disparados por Voltagem/efeitos dos fármacos
12.
J Mater Chem B ; 3(21): 4330-4337, 2015 Jun 07.
Artigo em Inglês | MEDLINE | ID: mdl-32262775

RESUMO

Multifunctional gold nanostructures effective at photothermal therapy (PTT) and high-quality X-ray/computed tomography (CT) imaging have drawn much attention in recent research. In particular, the development of improved PTT against cancer has been a particularly focused area of research for which the clinical need is great. Since the intracellular concentration of gold nanostructures is critical for their therapeutic photothermal efficacy, we decorated gold nanostructures with positively charged polyethylene glycol (PEG) to boost their degree of uptake by the cell. Herein gold nanostars (GNSs) were decorated with amine-terminated PEG (GNS-PEG-NH2) and methoxy-terminated PEG (GNS-mPEG). PEGylated GNSs showed good dispersivity, high stability and low cytotoxicity. Moreover, compared with GNS-mPEG, GNS-PEG-NH2 exhibited superior thermal therapeutic efficacy against breast tumor cells due to their higher cellular uptake. Measurement of the X-ray absorption coefficient revealed that the attenuation of GNS-PEG-NH2 was about 3.6-fold higher than that of the commercial CT contrast agent iodixanol at the concentration of 25 mg L-1. Importantly, GNS-PEG-NH2 also exhibited effective tumor therapeutic efficacy in vivo, and the tumor sites injected with GNS-PEG-NH2 showed high contrast X-ray/CT imaging. And most of the injected GNS-PEG-NH2 was cleared from tumors 15 days post-injection, indicating rapid clearance and minimal toxicity of GNS-PEG-NH2. Such PEGylated GNSs, when used for producing high-contrast images to guide enhanced PTT therapy, may thus provide new opportunities for the development of cancer theranostics.

13.
Analyst ; 140(3): 750-5, 2015 Feb 07.
Artigo em Inglês | MEDLINE | ID: mdl-25514163

RESUMO

Detection of malignant cells from malignant effusion is crucial to establish or adjust therapies of patients with cancer. The conventional qualitative detection in malignant pleuroperitoneal effusion is cytological analysis, which is time-consuming and complicated. Therefore, a faster and more convenient detection strategy is urgently needed. In this study, we report a rapid method to detect malignant cells from malignant pleuroperitoneal effusion (hydrothorax and ascites) of patients using IR-808, a tumor-targeted near-infrared (NIR) fluorescent heptamethine dye (tNRI dye), which exhibited superior labeling efficacy without specific conjugation to biomarkers. The targeted imaging performance toward malignant cells using IR-808 was confirmed by comparing with normal cells, and the fluorescence stability assay of IR-808 in malignant effusion was performed from 1 h to 48 h. In order to save time and dose, the incubation time and concentration were optimized to 10 min and 5 µM, which were used to detect malignant cells from 28 clinical samples of malignant pleuroperitoneal effusion. The results revealed that IR-808 could be internalized selectively by malignant cells of samples, and these malignant cells could be easily distinguished from normal cells under a fluorescence microscope. The positive rates between cytological analysis and the IR-808 staining method were 86% (24/28) and 79% (22/28), respectively. An excellent concordance level (Kappa = 0.752, P < 0.001) was observed between the two methods. Our results indicated that IR-808, a new NIR fluorescent heptamethine dye with unique optical imaging and tumor targeting properties, could provide a fast and simple way to detect a broad spectrum of malignant cells from malignant pleuroperitoneal effusion in patients.


Assuntos
Corantes Fluorescentes , Indóis , Neoplasias/diagnóstico , Peritônio/patologia , Derrame Pleural Maligno/diagnóstico , Espectroscopia de Luz Próxima ao Infravermelho/métodos , Células Cultivadas , Endotélio Vascular/citologia , Feminino , Corantes Fluorescentes/química , Humanos , Indóis/química , Masculino , Pessoa de Meia-Idade
14.
Yao Xue Xue Bao ; 47(10): 1336-40, 2012 Oct.
Artigo em Chinês | MEDLINE | ID: mdl-23289146

RESUMO

To investigate the effects of human anti-BAFF scFv-Fc against the hsBAFF, ICR mice were randomly divided into six groups: control, hsBAFF (1 mg x kg(-1)), hsBAFF (1 mg x kg(-1)) + Ab (1 mg x kg(-1)), hsBAFF (1 mg x kg(-1)) + Ab (2 mg x kg(-1)), hsBAFF (1 mg x kg(-1)) + human IgG (1 mg x kg(-1)) and hsBAFF (1 mg x kg(-1)) + human IgG (2 mg x kg(-1)) groups. The effects of scFv-Fc administration on the proliferation of B lymphocytes were evaluated using an MTT assay. The titres of antibody in the serum and B lymphocytes differentiation were assessed by ELISA and flow cytometry, respectively. The results showed that administration of scFv-Fc to mice injected with hsBAFF significantly prevented human BAFF-induced increases in splenic B cell numbers and serum immunoglobulin levels. Furthermore, this fully human antibody would avoid inducing the human anti-mouse antibody (HAMA) response when used in humans. These findings suggest that the compact antibody may be useful in therapeutic or diagnostic application of the BAFF-associated autoimmune diseases in human.


Assuntos
Fator Ativador de Células B/imunologia , Proliferação de Células , Fragmentos Fc das Imunoglobulinas/imunologia , Proteínas Recombinantes de Fusão/imunologia , Anticorpos de Cadeia Única/imunologia , Animais , Fator Ativador de Células B/metabolismo , Linfócitos B/citologia , Peso Corporal , Diferenciação Celular , Células Cultivadas , Feminino , Humanos , Fragmentos Fc das Imunoglobulinas/metabolismo , Imunoglobulina G/sangue , Imunoglobulina G/imunologia , Imunoglobulina M/sangue , Camundongos , Camundongos Endogâmicos ICR , Distribuição Aleatória , Proteínas Recombinantes de Fusão/metabolismo , Anticorpos de Cadeia Única/metabolismo , Baço/citologia
15.
Appl Biochem Biotechnol ; 157(3): 562-74, 2009 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-19099209

RESUMO

Elevated levels of B-cell-activating factor of the tumor necrosis factor family (BAFF) have been implicated in the pathogenesis of autoimmune diseases in human. In this study, we have constructed a vector for the expression of a novel compact antibody composed of anti-BAFF single-chain antibody fragment (scFv) and the Fc region (the hinge region, CH2, and CH3 domains) of human IgG1 in Chinese hamster ovary cells. The scFv-Fc fusion protein, showing spontaneous Fc fragment-mediated homodimerization via disulfide bridges, was affinity-purified on protein A Sepharose from culture supernatant. The scFv-Fc antibody was demonstrated to retain high binding affinity to antigen and prolonged clearance time in blood and to possess some human IgG crystallizable fragment effector functions such as protein A binding and antibody-dependent cellular cytotoxicity. These results suggest that this recombinant antibody may have therapeutic applications in the therapy of autoimmune disorders mediated by BAFF.


Assuntos
Fator Ativador de Células B/imunologia , Fragmentos Fc das Imunoglobulinas/imunologia , Fragmentos de Imunoglobulinas/imunologia , Proteínas Recombinantes de Fusão/imunologia , Animais , Fator Ativador de Células B/metabolismo , Western Blotting , Células CHO , Cricetinae , Cricetulus , Ensaio de Imunoadsorção Enzimática , Humanos , Fragmentos Fc das Imunoglobulinas/metabolismo , Fragmentos de Imunoglobulinas/metabolismo , Modelos Biológicos , Proteínas Recombinantes de Fusão/metabolismo
16.
Appl Microbiol Biotechnol ; 79(3): 423-31, 2008 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-18427805

RESUMO

Elevated levels of B-cell-activating factor of the TNF family (BAFF) have been implicated in the pathogenesis of autoimmune diseases in human. In this study, an anti-BAFF single-chain antibody fragment (scFv) was genetically linked to the C terminus of the enhanced green fluorescent protein (EGFP) to generate an EGFP/scFv fusion protein. The EGFP/scFv fusion protein had an apparent molecular weight of 52 kDa and was identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis and Western blot analysis. After being purified by immobilized metal affinity chromatography, the fusion protein exhibited similar fluorescence spectra with native EGFP. Enzyme-linked immunosorbent assay and fluorescence-activated cell sorting showed the EGFP/scFv could bind to human soluble BAFF and BAFF positive cell lines in vitro. The binding of EGFP/scFv can also be visualized under laser scanning confocal microscopy. Furthermore, the results of the competition assay indicated its antigen binding specificity. Therefore, the fusion protein EGFP/scFv has several characteristics including high sensitivity, stability and convenience for manipulation, and can be a powerful tool for the study of the underlying pathology of BAFF relevant to autoimmune diseases.


Assuntos
Anticorpos/imunologia , Antígenos/química , Antígenos/imunologia , Fator Ativador de Células B/química , Fator Ativador de Células B/imunologia , Especificidade de Anticorpos , Antígenos/genética , Antígenos/isolamento & purificação , Fator Ativador de Células B/genética , Fator Ativador de Células B/isolamento & purificação , Linhagem Celular , Ensaio de Imunoadsorção Enzimática , Expressão Gênica , Vetores Genéticos/genética , Proteínas de Fluorescência Verde/química , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/imunologia , Proteínas de Fluorescência Verde/isolamento & purificação , Humanos , Peso Molecular , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/imunologia , Proteínas Recombinantes de Fusão/isolamento & purificação
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