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1.
Zhongguo Zhong Yao Za Zhi ; 45(3): 623-630, 2020 Feb.
Artigo em Chinês | MEDLINE | ID: mdl-32237522

RESUMO

The aim of this paper was to investigate the effect of Faeces Bombycis(FB) on the intestinal microflora in rats with syndrome of damp retention in middle-jiao, and to explore its mechanism in regulating intestinal microflora from the perspective of microorganisms contained in FB. The contents of antidiuretic hormone(ADH) and C-reactive protein(CRP) in serum and aquaporin 3(AQP3) in jejunum were determined by enzyme-linked immunosorbent assay(ELISA). Illumina Miseq platform was used for high-throughput sequencing of the rat feces and FB. The ELISA results showed that as compared with the normal control group, the contents of ADH and CRP in the model group were significantly increased(P<0.05), and the content of AQP3 was significantly decreased(P<0.05). After drug administration, the ADH, CRP and AQP3 contents were recovered. Sequencing of rat feces showed that the ACE, Chao1 and Shannon indexes of the intestinal microflora were the lowest in the model group. As compared with the normal control group, the levels from phylum to genus were all significantly changed in model group, and Proteobacteria, Acinetobacter, Anaerobacter, Pseudomonas, and Parabacteroides levels were significantly increased(P<0.05), while Marvinbryantia level was significantly decreased(P<0.05). As compared with the model group, Proteobacteria was significantly decreased in the FB low and high dose groups(P<0.05), and Acinetobacter, Anaerobacter, Pseudomonas, Parabacteroides levels were significantly decreased in the low, medium and high dose groups(P<0.05), while Lachnoanaerobaculum, Intestinimonas and Marvinbryantia were increased significantly in the high dose group(P<0.05). Sequencing analysis of FB showed that the relative abundance of Leclercia, Pantoea, Brachybacterium, Shimwellia, Hartmannibacter, Klebsiella, Serratia, Aurantimonas, Paenibacillus and Bacillus was high in the FB, but they were basically not present or little in the rat feces. In conclusion, FB may play a role in the treatment of "syndrome of damp retention in middle-jiao" by balancing the intestinal microflora, and this effect may be related to the metabolites of microorganisms in the FB.


Assuntos
Bombyx/química , Fezes/química , Microbioma Gastrointestinal , Animais , Aquaporina 3/análise , Proteína C-Reativa/análise , Sequenciamento de Nucleotídeos em Larga Escala , Medicina Tradicional Chinesa , Ratos , Vasopressinas/sangue
2.
Artigo em Inglês | MEDLINE | ID: mdl-31931329

RESUMO

The silkworm, Bombyx mori, is a promising expression system for the production of recombinant proteins, but the purification of these proteins is not easy because of the large amount of host proteins present. To investigate purity, recovery and scale-up ability of the purification of recombinant proteins expressed in silkworm larval hemolymph without any affinity tags, we used mCherry, a red fluorescence protein, as a model. The host cell proteins could be greatly reduced using a three-step chromatography protocol consisting of hydrophobic interaction chromatography (HIC), size exclusion chromatography (SEC) and heparin chromatography after heat pretreatment. The thermal treatment had the greatest impact on the removal of host cell extracellular proteins and increasing purity. There were still some minor traces of host cell proteins in the purified sample, which showed that the purification of recombinant proteins from the silkworm hemolymph was still challenging. The proposed protocol and affinity tag purification reduced the overall protein content by 99.84% and 99.95%, respectively, while the amount of DNA was reduced by 98.41% and 99.53%, respectively. Purities of our proposed protocol based on SDS-PAGE and capillary electrophoresis (CE) analyses were 85.45% and 43.60%, respectively, while those of Strep-tag affinity purification were 100% or 63.69%, respectively. Using densitometry, the overall recovery was calculated was 5.78%, which was higher than 4.09% using Strep-tag affinity purification. This proposed protocol, mainly based on thermal treatment, HIC, SEC and HiTrap Heparin HP column chromatography, is applicable to an upscalable purification for the silkworm expression system without employing affinity tag chromatography process.


Assuntos
Bombyx/química , Cromatografia de Afinidade/métodos , Hemolinfa/química , Larva/química , Proteínas Recombinantes de Fusão/isolamento & purificação , Animais , Bombyx/metabolismo , Eletroforese em Gel de Poliacrilamida , Larva/metabolismo , Oligopeptídeos/genética , Oligopeptídeos/metabolismo , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/genética
3.
J Sci Food Agric ; 100(4): 1418-1425, 2020 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-31667852

RESUMO

BACKGROUND: Silkworm pupa oil polyunsaturated fatty acid (SPO PUFA) has been confirmed to have a cholesterol-lowering function. METHODS AND RESULTS: The effect of SPO PUFA and its main component, α-linolenic acid (ALA), on the metabolism of cholesterol and its regulation was investigated. The model of lipid denatured cells were constructed to carry out lipid accumulation, cholesterol metabolism and transformation. Real-time PCR and western blots were also used to analyze the expression levels of related genes and proteins to investigate the cholesterol efflux regulation mechanism. The data indicated that SPO PUFA and ALA dose-dependently decreased intracellular total cholesterol (TC) and enhanced total bile acid (TBA). They could also promote cholesterol removal by enhancing bile acid secretion and by upregulating genes LXRα, PPARγ, ABCA1, ABCG1, and CYP7A1, which were regulated by LXRα/PPARγ-ABCA1/ABCG1-CYP7A1 nuclear receptor signal pathways. CONCLUSIONS: This study is of great significance in maintaining the balance of cholesterol and lipid metabolism, and in reducing the risk of steatohepatitis. © 2019 Society of Chemical Industry.


Assuntos
Bombyx/química , Colesterol/metabolismo , Ácidos Graxos Insaturados/farmacologia , Hepatócitos/metabolismo , Pupa/química , Transportador 1 de Cassete de Ligação de ATP/genética , Transportador 1 de Cassete de Ligação de ATP/metabolismo , Membro 1 da Subfamília G de Transportadores de Cassetes de Ligação de ATP/genética , Membro 1 da Subfamília G de Transportadores de Cassetes de Ligação de ATP/metabolismo , Animais , Linhagem Celular , Colesterol 7-alfa-Hidroxilase/genética , Colesterol 7-alfa-Hidroxilase/metabolismo , Hepatócitos/efeitos dos fármacos , Humanos , Metabolismo dos Lipídeos/efeitos dos fármacos , PPAR gama/genética , PPAR gama/metabolismo , Transdução de Sinais/efeitos dos fármacos
4.
Insect Mol Biol ; 29(1): 104-111, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-31390480

RESUMO

Acetylation is an important, reversible posttranslational modification to a protein. In a previous study, we found that there were a large number of acetylated sites in various nutrient storage proteins of the silkworm haemolymph. In this study, we confirmed that acetylation can affect the stability of nutrient storage protein Bombyx mori apolipophorin-III (BmApoLp-III). First, the expression of BmApoLp-III could be upregulated when BmN cells were treated with the deacetylase inhibitor panobinostat (LBH589); similarly, the expression was downregulated when the cells were treated with the acetylase inhibitor C646. Furthermore, the increase in acetylation by LBH589 could inhibit the degradation and improve the accumulation of BmApoLp-III in BmN cells treated with cycloheximide and MG132 respectively. Moreover, we found that an increase in acetylation could decrease the ubiquitination of BmApoLp-III and vice versa; therefore, we predicted that acetylation could improve the stability of BmApoLp-III by competing for ubiquitination and inhibiting the protein degradation pathway mediated by ubiquitin. Additionally, BmApoLp-III had an antiapoptosis function that increased after LBH589 treatment, which might have been due to the improved protein stability after acetylation. These results have laid the foundation for further study on the mechanism of acetylation in regulating the storage and utilization of silkworm nutrition.


Assuntos
Apolipoproteínas/química , Bombyx/química , Proteínas de Insetos/química , Estabilidade Proteica/efeitos dos fármacos , Acetilação , Animais , Apolipoproteínas/metabolismo , Benzoatos/farmacologia , Bombyx/efeitos dos fármacos , Linhagem Celular , Cicloeximida/farmacologia , Proteínas de Insetos/metabolismo , Leupeptinas/farmacologia , Panobinostat/farmacologia , Pirazóis/farmacologia
5.
Artif Cells Nanomed Biotechnol ; 48(1): 68-76, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31852285

RESUMO

Mulberry silkworm cocoon (MSC) carbonisata has been used for the treatment of inflammatory diseases for hundreds of years; however, after years of research efforts, little information is available on its anti-inflammatory components and underlying mechanism. We developed novel carbon dots (CDs) derived from MSC carbonisata (MSC-CDs), for the first time, with an average diameter of 2.26-9.35 nm and a quantum yield (QY) of 6.32%. The MSC-CDs were prepared using a modified pyrolysis method, and no further modification and external surface passivation agent was required. With abundant surface groups, MSC-CDs showed distinct solubility and bioactivity. In this study, we innovatively used three classical experimental models of inflammation to evaluate the anti-inflammatory bioactivity of MSC-CDs. The results indicated that MSC-CDs exhibited marked anti-inflammatory bioactivity which was likely mediated by inhibition of the expression of interleukin-6 and tumour necrosis factor-α. These results suggest that MSC-CDs possess a remarkable anti-inflammatory property, which provides evidence to support further investigation of the considerable potential and effective material basis of this traditional Chinese medicine.


Assuntos
Anti-Inflamatórios/química , Anti-Inflamatórios/farmacologia , Bombyx/química , Carbono/química , Carbono/farmacologia , Pontos Quânticos/química , Animais , Anti-Inflamatórios/uso terapêutico , Carbono/uso terapêutico , Edema/tratamento farmacológico , Feminino , Camundongos , Células RAW 264.7 , Sepse/tratamento farmacológico
6.
Molecules ; 24(21)2019 Oct 31.
Artigo em Inglês | MEDLINE | ID: mdl-31683683

RESUMO

Silk fibroin (SF) produced by the domesticated wild silkworm, Samia cynthia ricini (S. c. ricini) is attracting increasing interest owing to its unique mechanical properties, biocompatibility, and abundance in nature. However, its utilization is limited, largely due to lack of appropriate processing strategies. Various strategies have been assessed to regenerate cocoon SF, as well as the use of aqueous liquid fibroin (LFaq) prepared by dissolution of silk dope obtained from the silk glands of mature silkworm larvae in water. However, films cast from these fibroin solutions in water or organic solvents are often water-soluble and require post-treatment to render them water-stable. Here, we present a strategy for fabrication of water-stable films from S. c. ricini silk gland fibroin (SGF) without post-treatment. Aqueous ethanol induced gelation of fibroin in the posterior silk glands (PSG), enabling its separation from the rest of the silk gland. When dissolved in 1,1,1,3,3,3-hexafluoro-2-propanol (HFIP), the SGF-gel gave a solution from which a transparent, flexible, and water-insoluble film (SGFHFIP) was cast. Detailed structural characterization of the SGFHFIP as-cast film was carried out and compared to a conventional, water-soluble film cast from LFaq. FTIR and 13C solid-state NMR analyses revealed both cast films to be α-helix-rich. However, gelation of SGF induced by the 40%-EtOH-treatment resulted in an imperfect ß-sheet structure. As a result, the SGF-gel was soluble in HFIP, but some ß-sheet structural memory remains, and the SGFHFIP as-cast film obtained has some ß-sheet content which renders it water-resistant. These results reveal a structure water-solubility relationship in S. c. ricini SF films that may offer useful insights towards tunable fabrication of novel biomaterials. A plausible model of the mechanism that leads to the difference in water resistance of the two kinds of α-helix-rich films is proposed.


Assuntos
Bombyx/química , Fibroínas/química , Propanóis/química , Água/química , Aminoácidos/análise , Animais , Varredura Diferencial de Calorimetria , Espectroscopia de Ressonância Magnética Nuclear de Carbono-13 , Conformação Proteica em alfa-Hélice , Solubilidade , Soluções , Espectroscopia de Infravermelho com Transformada de Fourier , Termogravimetria , Difração de Raios X
7.
Molecules ; 24(20)2019 Oct 21.
Artigo em Inglês | MEDLINE | ID: mdl-31640173

RESUMO

Bombyx batryticatus is a well-known animal in traditional Chinese medicine. The aim of the research was to reveal the quality formation mechanism of B. batryticatus and to screen out the characteristic component used for the quality control. The anticonvulsant effects of B. batryticatus with a stiff time of one, five, and nine days (D1, D5 and D9, respectively) and healthy silkworm of the same developmental stage (SW) were determined by animal experiment. The dynamic changes in chemical composition were analyzed using UPLC-Q-TOF-MS-based metabolomics. D5 and D9 B. batryticatus exhibited significant anticonvulsant effects (p < 0.05 and p < 0.01, respectively). Accordingly, principal component analysis (PCA) and partial least squares discrimination analysis (PLS-DA) indicated that the chemical composition of D5 and D9 B. batryticatus changed significantly. The different metabolites mainly consisted of primary metabolites such as lipids and amino acids and secondary metabolites such as flavonoids, beauvericin, and glycolipids. Interestingly, the relative abundance of quercetin-7-O-ß-d-4-O-methylglucoside, the characteristic component of B. batryticatus, increased with stiff time and was promised to be used as an index component of quality control. The results expand our understanding of the quality formation mechanism of B. batryticatus. In addition, it highlights the potential of UPLC-Q-TOF-MS-based metabolomics for the quality control purpose of TCMs.


Assuntos
Bombyx/fisiologia , Metabolômica/métodos , Metilglucosídeos/análise , Animais , Anticonvulsivantes , Bombyx/química , Bombyx/microbiologia , Cromatografia Líquida de Alta Pressão , Análise dos Mínimos Quadrados , Espectrometria de Massas , Metilglucosídeos/química , Análise de Componente Principal , Quercetina , Metabolismo Secundário
8.
Molecules ; 24(20)2019 Oct 17.
Artigo em Inglês | MEDLINE | ID: mdl-31627317

RESUMO

Formation of the α-helical conformation in the poly-l-alanine (PA) sequence regions, subsequent structural transition to ß-sheet during natural spinning, and presence of residual α-helices in Samia cynthia ricini (S. c. ricini) native silk fiber have been experimentally proven. However, the aggregation state of the residual α-helices, and their influence on the mechanical deformation behavior in native fiber remain unclear. Here we show that the α-helices form an ordered aggregation state with a hexagonal packing in the aqueous solution, some of which remain during natural spinning. X-ray scattering and differential scanning calorimetry (DSC) analyses revealed occurrence of a structural transition of the residual α-helices to the ß-sheet structure, accompanied by disappearance of the plateau region in the force-strain curve, due to heat-treatment at ~220 °C. On the basis of X-ray scattering before and after tensile stretching of S. c. ricini native silk, a direct connection between the plateau region and the α-helix to ß-sheet structural transition was confirmed. Our findings demonstrate the importance of the PA sequence regions in fiber structure formation and their influence on the tensile deformation behavior of S. c. ricini silk, features believed to be essentially similar in other saturniid silks. We strongly believe the residual ordered α-helices to be strategically and systematically designed by S. c. ricini silkworms to impart flexibility in native silk fiber. We anticipate that these knowledge forms a basis for fruitful strategies in the design and development of amino acid sequences for artificial silks with desired mechanical properties.


Assuntos
Bombyx/química , Fibroínas/ultraestrutura , Peptídeos/química , Agregados Proteicos , Animais , Bombyx/fisiologia , Fibroínas/isolamento & purificação , Temperatura Alta , Larva/química , Larva/fisiologia , Teste de Materiais , Peptídeos/isolamento & purificação , Maleabilidade , Conformação Proteica em alfa-Hélice , Conformação Proteica em Folha beta , Resistência à Tração
9.
Int J Mol Sci ; 20(19)2019 Sep 27.
Artigo em Inglês | MEDLINE | ID: mdl-31569598

RESUMO

The interests of developing antimicrobial biomaterials based on silk sericin from Bombyx mori cocoon, have been shooting up in the last decades. Sericin is a valuable natural protein owing to its hydrophilicity, biodegradability, and biocompatibility. Here, we fabricated a sponge with antibacterial capacities for potential wound dressing application. By co-blending of sericin, polyvinyl alcohol (PVA) and zinc oxide nanoparticles (ZnONPs), the ZnONPs-sericin/PVA composite sponge (ZnONPs-SP) was successfully prepared after freeze-drying. Scanning electron microscopy showed the porous structure of ZnONPs-SP. Energy dispersive spectroscopy indicated the existence of Zn in the sponge. X-ray diffractometry revealed the hexagonal wurtzite structure of ZnONPs. Fourier transform infrared spectroscopy showed the biologic coupling of ZnONPs and sericin resulted in a decrease of α-helix and random coil contents, and an increase of ß-sheet structure in the sponge. The swelling experiment suggested ZnONPs-SP has high porosity, good hydrophilicity, and water absorption capability. The plate bacterial colony counting coupled with growth curve assays demonstrated that the composite sponge has an efficiently bacteriostatic effect against Staphylococcus aureus and Escherichia coli, respectively. Furthermore, the cell compatibility analysis suggested the composite sponge has excellent cytocompatibility on NIH3T3 cells. In all, ZnONPs-SP composite sponge has significant potentials in biomaterials such as wound dressing and tissue engineering.


Assuntos
Antibacterianos/química , Antibacterianos/farmacologia , Nanopartículas Metálicas/química , Polivinil/química , Polivinil/farmacologia , Sericinas/química , Óxido de Zinco/química , Animais , Bombyx/química , Sobrevivência Celular , Nanopartículas Metálicas/ultraestrutura , Camundongos , Testes de Sensibilidade Microbiana , Células NIH 3T3 , Porosidade , Espectroscopia de Infravermelho com Transformada de Fourier , Difração de Raios X
10.
Chemosphere ; 236: 124379, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31545189

RESUMO

In order to study the role of mulberry (Moms alba L) as an economic crop for remediation of cadmium (Cd) contaminated soil, the transport of Cd from mulberry to silkworm were investigated. Three varieties of mulberry (Yuesang-11, Nongsang-14, and Qiangsang-1) with three planting densities were cultivated in two heavy metal-contaminated fields named Dongkou in Shaoyang city and Linxiang in Yueyang city in Hunan province respectively. The both field soils were contaminated by heavy metals, especially by Cd. The potential risks of heavy metals in Linxiang's soil were higher than those in Dongkou's because of higher concentrations of Cd. Since the promotion of Cd concentrations in aerial parts (stem, branch and leaf) resulted from the increase of planting density, the method of high planting density is beneficial to improve the efficiency of the remediation of Cd contaminated soil. The percentages of average Cd contents of mulberry in Dongkou accounted for 44%, 20%, 18% and 16% in roots, stems, branches and leaves respectively, while the Cd contents were 38%, 27%, 19% and 16% distributed in roots, stems, branches and leaves respectively. Mulberry leaves from contaminated soils was applied in food source of silkworms in this study. Although there is Cd uptake occurred in silkworm growth and its products (cocoons and chrysalis), Cd contents in cocoons are lower than the national standard (100 µg*kg-1) for textile industry of China. Therefore, mulberry can be regarded as an economical crop to control soil contamination with Cd.


Assuntos
Cádmio/metabolismo , Recuperação e Remediação Ambiental/métodos , Morus/metabolismo , Poluentes do Solo/metabolismo , Animais , Biodegradação Ambiental , Bombyx/química , Bombyx/metabolismo , Cádmio/análise , China , Recuperação e Remediação Ambiental/economia , Frutas/química , Frutas/metabolismo , Morus/química , Morus/classificação , Folhas de Planta/química , Folhas de Planta/metabolismo , Raízes de Plantas/química , Raízes de Plantas/metabolismo , Poluentes do Solo/análise
11.
BMC Complement Altern Med ; 19(1): 241, 2019 Sep 05.
Artigo em Inglês | MEDLINE | ID: mdl-31488109

RESUMO

BACKGROUND: Mulberry silkworm larvae (Bombyx mori) are known as the oldest resource of food and traditional medicine. Although silkworm larvae have been reported to treat various chronic diseases, the effect of fermentation by microorganisms improving the biological activities of silkworm larvae was not reported. In the present study, fermented silkworm larvae was developed via solid-state fermentation with Aspergillus kawachii and investigated its anti-cancer activity in human hepatocellular carcinoma cells. METHODS: We investigated the anti-cancer effects of unfermented (SEE) and fermented silkworm larva ethanol extract (FSEE) on HepG2 human hepatocellular carcinoma cells as well as compared changes in free amino acid, fatty acid, and mineral contents. Anti-cancer activities were evaluated by SRB staining, cell cycle analysis, Annexin V staining, Hoechst staining, DNA fragmentation analysis and western blot analysis. Fatty acid, free amino acid and mineral contents of SEE and FSEE were determined by gas chromatography, amino acid analyzer and flame atomic absorption spectrophotometer, respectively. RESULTS: Compared with SEE, treatment with FSEE resulted in apoptotic cell death in HepG2 cells characterized by G0/G1 phase cell cycle arrest, DNA fragmentation, and formation of apoptotic bodies. Furthermore, FSEE significantly up-regulated pro-apoptotic as well as down-regulated anti-apoptotic proteins in HepG2 cells. However, an equivalent concentration of SEE did not induce cell cycle arrest or apoptosis in HepG2 cells. Moreover, fermentation process by Aspergillus kawachii resulted in enhancement of fatty acid contents in silkworm larvae, whereas amino acid and mineral contents were decreased. CONCLUSION: Collectively, this study demonstrates that silkworm larvae solid state-fermented by Aspergillus kawachii strongly potentiates caspase-dependent and -independent apoptosis pathways in human hepatocellular carcinoma cells by regulating secondary metabolites.


Assuntos
Antineoplásicos/farmacologia , Aspergillus/metabolismo , Bombyx/microbiologia , Carcinoma Hepatocelular/tratamento farmacológico , Larva/química , Neoplasias Hepáticas/tratamento farmacológico , Animais , Antineoplásicos/química , Antineoplásicos/metabolismo , Apoptose/efeitos dos fármacos , Bombyx/química , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/fisiopatologia , Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Fragmentação do DNA , Fermentação , Células Hep G2 , Humanos , Larva/microbiologia , Neoplasias Hepáticas/fisiopatologia
12.
Mater Sci Eng C Mater Biol Appl ; 104: 109890, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31500018

RESUMO

A protein-polymer blend system based on silkworm silk fibroin (SF) and polylactic acid (PLA) was systematically investigated to understand the interaction and miscibility of proteins and synthetic biocompatible polymers in the macro- and micro-meter scales, which can dramatically control the cell responses and enzyme biodegradation on the biomaterial interface. Silk fibroin, a semicrystalline protein with beta-sheet crystals, provides controllable crystal content and biodegradability; while noncrystallizable PDLLA provides hydrophobicity and thermal stability in the system. Differential scanning calorimetry (DSC) combined with scanning electron microscope (SEM) showed that the morphology of the blend films was uniform on a macroscopic scale, yet with tunable micro-phase patterns at different mixing ratios. Fourier transform infrared analysis (FTIR) revealed that structures of the blend system, such as beta-sheet crystal content, gradually changed with the mixing ratios. All blended samples have better stability than pure SF and PLA samples as evidenced by thermogravimetric analysis. Protease XIV enzymatic study showed that the biodegradability of the blend samples varied with their blending ratios and microscale morphologies. Significantly, the topology of the micro-phase patterns on the blends can promote cell attachment and manipulate the cell growth and proliferation. This study provided a useful platform for understanding the fabrication strategies of protein-synthetic polymer composites that have direct biomedical and green chemistry applications.


Assuntos
Fibroínas/química , Poliésteres/química , Polímeros/química , Seda/química , Animais , Materiais Biocompatíveis/química , Bombyx/química , Varredura Diferencial de Calorimetria/métodos , Microscopia Eletrônica de Varredura/métodos , Espectroscopia de Infravermelho com Transformada de Fourier/métodos
13.
J Sci Food Agric ; 99(15): 7008-7015, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31435932

RESUMO

BACKGROUND: Silkworm droppings have long been used in traditional medicine to remedy allergic itching, palsy, blood circulation problems, and arthritis in Asian countries. To investigate the anti-allergic effect of silkworm dropping extract (SDE) and its mechanism, we used a mouse model of food allergy induced by ovalbumin (OVA). RESULTS: SDE ameliorated the symptoms of OVA-induced food allergies, and the levels of T helper 2 (Th2)-related cytokines [such as interleukin (IL)-4, IL-5, IL-10, and IL-13] were found to be significantly decreased in both the spleen and mesenteric lymph nodes by SDE. Furthermore, SDE treatment directly inhibited OVA permeation, IL-4 production, and degranulation of mast cells; in contrast, immunoglobulin E (IgE) production from B cells was not affected. CONCLUSION: These results suggest that SDE has potential anti-allergic activities, and SDE may be useful in the treatment/prevention of allergic disorders such as food allergies, serving as therapeutic agents. © 2019 Society of Chemical Industry.


Assuntos
Antialérgicos/administração & dosagem , Bombyx/química , Fezes/química , Hipersensibilidade Alimentar/tratamento farmacológico , Células Th2/efeitos dos fármacos , Animais , Antialérgicos/química , Feminino , Hipersensibilidade Alimentar/genética , Hipersensibilidade Alimentar/imunologia , Humanos , Imunoglobulina E/imunologia , Interleucina-13/genética , Interleucina-13/imunologia , Interleucina-4/genética , Interleucina-4/imunologia , Interleucina-5/genética , Interleucina-5/imunologia , Mastócitos/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Ovalbumina/efeitos adversos , Células Th2/imunologia
14.
Biosens Bioelectron ; 142: 111595, 2019 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-31425973

RESUMO

The present work reports cocoon silk fibroin (SF)as a unique precursor for the in-situ fabrication of well-engineered, stable and leach free gold nanoparticle doped carbonaceous materials (AuNPs@NSC). In principle, at the molecular level, SF has a singular structure that can be converted to a N-doped aromatic carbon structure by heat treatment. The electrochemical properties of the prepared nanocomposite were examined by cyclic voltammetry and differential pulse voltammetry. A flexible three electrode sensor system with AuNPs@NSC-modified working electrodes has been developed, to achieve easy operation and quick and accurate responses. The electrochemical results showed that the sensor made by the AuNPs@NSC-modified working electrode demonstrated high sensitivity for the detection of rutin, which is attributed to the good distribution of the AuNPs on the carbon matrix. Using differential pulse voltammetry (DPV), the AuNPs@NSC electrode was found to have a linear response in the range of 0.11-250 µM and a comparably low limit of detection of 0.02 µM (S/N = 3). To ensure the accuracy and applicability of the sensors, the concentration of rutin in the commodity (rutin capsule, 10 mg/capsule) was examined, and the sensor provided high precision with a minimum relative error (RE) of 3.3%. These findings suggest that AuNPs@NSC can be considered to be a potential electrode material for the development of electrochemical devices and has great potential in extending their application to the flexible sensor field.


Assuntos
Técnicas Biossensoriais/instrumentação , Carbono/química , Fibroínas/química , Ouro/química , Nanocompostos/química , Rutina/análise , Animais , Bombyx/química , Cápsulas , Técnicas Eletroquímicas/instrumentação , Eletrodos , Desenho de Equipamento , Limite de Detecção , Nanopartículas Metálicas/química
15.
J Phys Chem Lett ; 10(15): 4278-4284, 2019 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-31318218

RESUMO

Silk continues to amaze: over the past decade, new research threads have emerged that include the use of silk fibroin for advanced pharmaceutics, including its suitability for drug delivery. Despite this ongoing interest, the details of silk fibroin structures and their subsequent drug interactions at the molecular level remain elusive, primarily because of the difficulties encountered in modeling the silk fibroin molecule. Here, we generated an atomistic silk model containing amorphous and crystalline regions. We then exploited advanced well-tempered metadynamics simulations to generate molecular conformations that we subsequently exposed to classical molecular dynamics simulations to monitor both drug binding and release. Overall, this study demonstrated the importance of the silk fibroin primary sequence, electrostatic interactions, hydrogen bonding, and higher-order conformation in the processes of drug binding and release.


Assuntos
Doxorrubicina/química , Portadores de Fármacos/química , Fibroínas/química , Animais , Bombyx/química , Cristalização , Liberação Controlada de Fármacos , Ligação de Hidrogênio , Simulação de Dinâmica Molecular , Conformação Proteica , Eletricidade Estática , Termodinâmica , Água/química
16.
J Microencapsul ; 36(5): 432-446, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31238752

RESUMO

The present study was aimed at preparing and evaluating levocetirizine (LCZD) loaded emulgel containing tamanu oil and sericin for atopic dermatitis (AD) therapy. The emulgel envisaged topical delivery of LCZD utilising natural antioxidants for superior therapeutic outcomes when compared with other conventional therapy. Tamanu oil based microemulsion (ME) was optimised utilising Box-Behnken design (BBD). The OPT-ME displayed globule size 379.5 ± 2.33 nm, polydispersity index 0.284, drug loading 0.41 ± 0.01% w/w, entrapment efficiency 94.34 ± 2.11% w/w and drug release 86.24 ± 4.90% respectively over a period of 24 h. The optimised formulation (OPT-ME) was further incorporated into sericin gel to form emulgel (LSE). In vivo pharmacodynamic studies revealed enhanced therapeutic potential of emulgel in terms of reduced scratching frequency and erythema score when compared with conventional gel. The superior therapeutic potential was further witnessed through histopathological and biochemical studies. The emulgel can be an alternative appropriate dosage form for the treatment of AD.


Assuntos
Cetirizina/administração & dosagem , Dermatite Atópica/tratamento farmacológico , Emulsões/química , Óleos Vegetais/química , Sericinas/química , Animais , Bombyx/química , Calophyllum/química , Cetirizina/farmacocinética , Cetirizina/uso terapêutico , Chlorocebus aethiops , Dermatite Atópica/induzido quimicamente , Dermatite Atópica/patologia , Dinitroclorobenzeno , Sistemas de Liberação de Medicamentos , Liberação Controlada de Fármacos , Feminino , Masculino , Ratos Wistar , Absorção Cutânea , Células Vero
17.
Int J Mol Sci ; 20(12)2019 Jun 21.
Artigo em Inglês | MEDLINE | ID: mdl-31234286

RESUMO

Silk fiber has become a research focus because of its comprehensive mechanical properties. Metal ions can influence the conformational transition of silk fibroin. Current research is mainly focused on the role of a single ion, rather than the whole metal ion environment. Here, we report the effects of the overall metal ion environment on the secondary structure and mechanical properties of silk fibers after direct injection and feeding of silkworms with EDTA. The metal composition of the hemolymph, silk gland, and silk fiber changed significantly post EDTA treatment. Synchrotron FTIR analysis indicated that the secondary structure of silk fiber after EDTA treatment changed dramatically; particularly, the ß-sheets decreased and the ß-turns increased. Post EDTA treatment, the silk fiber had significantly decreased strength, Young's modulus, and toughness as compared with the control groups, while the strain exhibited no obvious change. These changes can be attributed to the change in the metal ion environment in the silk fibroin and sericin in the silk gland. Our investigation provides a new theoretical basis for the natural silk spinning process, and our findings could help develop a method to modify the mechanical properties of silk fiber using metal ions.


Assuntos
Bombyx/metabolismo , Ácido Edético/metabolismo , Metais/metabolismo , Seda/química , Ração Animal/análise , Animais , Fenômenos Biomecânicos , Bombyx/química , Ácido Edético/administração & dosagem , Módulo de Elasticidade , Metais/análise , Conformação Proteica em Folha beta , Seda/metabolismo , Espectroscopia de Infravermelho com Transformada de Fourier , Estresse Mecânico
18.
Oxid Med Cell Longev ; 2019: 7897584, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31198493

RESUMO

Bombyx batryticatus is a known traditional Chinese medicine (TCM) utilized to treat convulsions, epilepsy, cough, asthma, headaches, and purpura in China for thousands of years. This study is aimed at investigating the antiepileptic effects of protein-rich extracts from Bombyx batryticatus (BBPs) on seizure in mice and exploring the protective effects of BBPs against H2O2-induced oxidative stress in PC12 cells and their underlying mechanisms. Maximal electroshock-induced seizure (MES) and pentylenetetrazole- (PTZ-) induced seizure in mice and the histological analysis were carried out to evaluate the antiepileptic effects of BBPs. The cell viability of PC12 cells stimulated by H2O2 was determined by MTT assay. The apoptosis and ROS levels of H2O2-stimulated PC12 cells were determined by flow cytometry analysis. Furthermore, the levels of malondialdehyde (MDA), superoxide dismutase (SOD), lactate dehydrogenase (LDH), and glutathione (GSH) in PC12 cells were assayed by ELISA and expressions of caspase-3, caspase-9, Bax, Bcl-2, PI3K, Akt, and p-Akt were evaluated by Western blotting and quantitative real-time polymerase chain reaction (RT-qPCR) assays. The results revealed that BBPs exerted significant antiepileptic effects on mice. In addition, BBPs increased the cell viability of H2O2-stimulated PC12 cells and reduced apoptotic cells and ROS levels in H2O2-stimulated PC12 cells. By BBPs treatments, the levels of MDA and LDH were reduced and the levels of SOD and GSH-Px were increased in H2O2-stimulated PC12 cells. Moreover, BBPs upregulated the expressions of PI3K, Akt, p-Akt, and Bcl-2, whereas they downregulated the expressions of caspase-9, caspase-3, and Bax in H2O2-stimulated PC12 cells. These findings suggested that BBPs possessed potential antiepileptic effects on MES and PTZ-induced seizure in mice and protective effects on H2O2-induced oxidative stress in PC12 cells by exerting antioxidative and antiapoptotic effects via PI3K/Akt signaling pathways.


Assuntos
Anticonvulsivantes/farmacologia , Bombyx/química , Regulação da Expressão Gênica/efeitos dos fármacos , Proteínas de Insetos/administração & dosagem , Fármacos Neuroprotetores/farmacologia , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Convulsões/tratamento farmacológico , Animais , Anticonvulsivantes/administração & dosagem , Antioxidantes/farmacologia , Apoptose , Sobrevivência Celular , Convulsivantes/toxicidade , Eletrochoque/efeitos adversos , Peróxido de Hidrogênio/toxicidade , Proteínas de Insetos/farmacologia , Masculino , Malondialdeído/metabolismo , Camundongos , Fármacos Neuroprotetores/administração & dosagem , Oxidantes/toxicidade , Estresse Oxidativo/efeitos dos fármacos , Células PC12 , Pentilenotetrazol/toxicidade , Fosfatidilinositol 3-Quinases/genética , Proteínas Proto-Oncogênicas c-akt/genética , Ratos , Convulsões/etiologia , Convulsões/metabolismo , Convulsões/patologia , Transdução de Sinais
19.
Carbohydr Polym ; 216: 17-24, 2019 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-31047054

RESUMO

The cutaneous tissue contains cellular protein and polysaccharide components which together maintain the functionality of the tissue. In this study, silk fibroin (SF) and konjac glucomannan (KGM) were physically crosslinked to form biocompatible protein/polysaccharide sponges with tunable mechanical properties for wound dressing application. The pore structure of sponges can be adjusted by changing blend ratio of SF/KGM, forming homogeneous interconnected pore structure. FTIR and Raman results revealed the intermolecular interaction between SF and KGM, suggesting the formation of interpenetrating polymer network after ethanol/ammonium hydroxide treatment. Raising KGM content significantly enhanced water-absorption, water-retention abilities, and compression strength of porous sponges. Especially, the composite sponges possessed a similar compressive modulus with native skin tissue, showing a matched flexibility for wound treatment. Moreover, the cell viability results based on human dermal fibroblast cells demonstrated that the sponge showed excellent biocompatibility for cell adhesion and proliferation. Therefore, due to the strong water-absorption capacity, moist environment, similar compressive modulus with skin tissue and excellent biocompatibility, the composite sponges have potential application in wound dressings.


Assuntos
Materiais Biocompatíveis/química , Fibroínas/química , Mananas/química , Curativos Oclusivos , Absorção Fisico-Química , Animais , Materiais Biocompatíveis/toxicidade , Bombyx/química , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Força Compressiva , Módulo de Elasticidade , Fibroblastos/metabolismo , Fibroínas/toxicidade , Humanos , Mananas/toxicidade , Porosidade , Água/química
20.
Colloids Surf B Biointerfaces ; 180: 118-126, 2019 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-31035055

RESUMO

With the increase of vascular diseases in recent years, it is of importance to develop an anti-occlusion stent graft, which can meet the requirements of transplants for a long term. In this paper, we describe a silk fibroin (SF)/heparin-functionalized bifurcated stent graft (BSG) using textile forming technology. The BSGs were prototyped based on seamless weaving technology, and the surface was modified with SF-loaded heparin under steam/air treatment to improve their patency. The physical properties such as thickness, water permeability, contact angle, mechanical properties, and in vitro drug release and coagulation time of the BSGs were examined. The results showed that heparin modification can improve its coagulation time, and the water permeability resistance of the BSGs reached 1.154 ± 0.854 mL/(cm2×min), while their thicknesses were just 0.085 ± 0.004 mm. The heparin release of the BSGs showed that the release time was prolonged upon steam treatment by means of the increase in the ß-sheet structure and crystallinity of SF. The viability and attachment of human vascular smooth muscle (HVSM) cells cultured in the release of modified BSGs demonstrated that the modified BSGs could significantly inhibit the proliferation of HVSM cells. The heparin-functionalized BSG with satisfactory thickness, water permeability resistance and anti-occlusion function, which has potential applications in the treatment of vascular diseases.


Assuntos
Prótese Vascular , Preparações de Ação Retardada/química , Fibroínas/química , Heparina/química , Animais , Bombyx/química , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Preparações de Ação Retardada/farmacologia , Liberação Controlada de Fármacos , Fibroínas/isolamento & purificação , Heparina/farmacologia , Humanos , Interações Hidrofóbicas e Hidrofílicas , Cinética , Miócitos de Músculo Liso/citologia , Miócitos de Músculo Liso/efeitos dos fármacos , Conformação Proteica em Folha beta , Tecidos Suporte
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