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1.
Int J Mol Sci ; 22(4)2021 Feb 23.
Artigo em Inglês | MEDLINE | ID: mdl-33672103

RESUMO

Hydrogen sulfide (H2S) has long been considered as a toxic gas, but as research progressed, the idea has been updated and it has now been shown to have potent protective effects at reasonable concentrations. H2S is an endogenous gas signaling molecule in mammals and is produced by specific enzymes in different cell types. An increasing number of studies indicate that H2S plays an important role in cardiovascular homeostasis, and in most cases, H2S has been reported to be downregulated in cardiovascular diseases (CVDs). Similarly, in preclinical studies, H2S has been shown to prevent CVDs and improve heart function after heart failure. Recently, many H2S donors have been synthesized and tested in cellular and animal models. Moreover, numerous molecular mechanisms have been proposed to demonstrate the effects of these donors. In this review, we will provide an update on the role of H2S in cardiovascular activities and its involvement in pathological states, with a special focus on the roles of exogenous H2S in cardiac protection.

2.
J Alzheimers Dis ; 80(2): 787-797, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33579846

RESUMO

BACKGROUND: In recent years, the efficacy of type 2 diabetes mellitus (T2DM) drugs in the treatment of Alzheimer's disease (AD) has attracted extensive interest owing to the close associations between the two diseases. OBJECTIVE: Here, we screened traditional Chinese medicine (TCM) and multi-target ingredients that may have potential therapeutic effects on both T2DM and AD from T2DM prescriptions. METHODS: Network pharmacology and molecular docking were used. RESULTS: Firstly, the top 10 frequently used herbs and corresponding 275 active ingredients were identified from 263 T2DM-related TCM prescriptions. Secondly, through the comparative analysis of 208 potential targets of ingredients, 1,740 T2DM-related targets, and 2,060 AD-related targets, 61 common targets were identified to be shared. Thirdly, by constructing pharmacological network, 26 key targets and 154 representative ingredients were identified. Further enrichment analysis showed that common targets were involved in regulating multiple pathways related to T2DM and AD, while network analysis also found that the combination of Danshen (Radix Salviae)-Gancao (Licorice)-Shanyao (Rhizoma Dioscoreae) contained the vast majority of the representative ingredients and might be potential for the cotreatment of the two diseases. Fourthly, MAPK1, PPARG, GSK3B, BACE1, and NR3C1 were selected as potential targets for virtual screening of multi-target ingredients. Further docking studies showed that multiple natural compounds, including salvianolic acid J, gancaonin H, gadelaidic acid, icos-5-enoic acid, and sigmoidin-B, exhibited high binding affinities with the five targets. CONCLUSION: To summarize, the present study provides a potential TCM combination that might possess the potential advantage of cotreatment of AD and T2DM.

3.
J Appl Microbiol ; 2021 Feb 11.
Artigo em Inglês | MEDLINE | ID: mdl-33570819

RESUMO

Obesity is considered as a risk factor for chronic health diseases such as heart diseases, cancer and diabetes 2. Reduced physical activities, lifestyle, poor nutritional diet and genetics are among the risk factors associated with the development of obesity. In recent years, several studies have explored the link between the gut microbiome and the progression of diseases including obesity, with the shift in microbiome abundance and composition being the main focus. The alteration of gut microbiome composition affects both nutrients metabolism and specific gene expressions, thereby disturbing body physiology. Specifically, the abundance of fibre-metabolizing microbes is associated with weight loss and that of protein and fat-metabolizing bacteria with weight gain. Various internal and external factors such as genetics, maternal obesity, mode of delivery, breastfeeding, nutrition, antibiotic use and the chemical compounds present in the environment are known to interfere with the richness of the gut microbiota (GM), thus influencing weight gain/loss and ultimately the development of obesity. However, the effectiveness of each factor in potentiating the shift in microbes' abundance to result in significant changes that can lead to obesity is not yet clear. In this review, we will highlight the factors involved in shaping GM, their influence on obesity and possible interventions. Understanding the influence of these factors on the diversity of the GM and how to improve their effectiveness on disease conditions could be keys in the treatment of metabolic diseases.

4.
J Diabetes Res ; 2020: 2817972, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33062708

RESUMO

Diabetes is prevalent worldwide, but ideally intensive therapeutic strategy in clinical diabetes and diabetic nephropathy (DN) is still lack. Pyruvate is protective from glucometabolic disturbances and kidney dysfunction in various pathogenic insults. Present studies focused on oral pyruvate effects on diabetes status and DN with 0.35% pyruvate in pyruvate-enriched oral rehydration solution (Pyr-ORS) and 1% pyruvate as drinking water for 8 weeks, using the model of diabetic db/db mice. Both Pyr-ORS and 1% pyruvate showed comparable therapeutic effectiveness with controls of body weight and blood sugar, increases of blood insulin levels, and improvement of renal function and pathological changes. Aberrant key enzyme activities in glucometabolic pathways, AR, PK, and PDK/PDH, were also restored; indexes of oxidative stress and inflammation, NAD+/NADH ratio, and AGEs in the kidneys were mostly significantly preserved after pyruvate treatments. We concluded that oral pyruvate delayed DN progression in db/db mice and the modified Pyr-ORS formula might be an ideal novel therapeutic drink in clinical prevention and treatment of type 2 diabetes and DN.

5.
FEBS Open Bio ; 10(5): 827-834, 2020 05.
Artigo em Inglês | MEDLINE | ID: mdl-32150786

RESUMO

Endoplasmic reticulum (ER) stress plays a critical role in the development of diabetic nephropathy (DN). We previously demonstrated that pyruvate (Pyr)-enriched oral rehydration solution improved glucometabolic disorders and ameliorated DN outcome in db/db mice. Here, we investigated the effects of Pyr on high glucose-induced ER stress and apoptosis in HK-2 cells. Our results suggest that high glucose can induce reactive oxygen species production, apoptosis and ER stress in HK-2 cells, and that Pyr treatment can ameliorate these effects and restore the expression of key proteins involved in ER stress. Thus, Pyr may have potential for the development of novel strategies for the prevention and treatment of clinical DN.

6.
J Am Heart Assoc ; 8(1): e009146, 2019 01 08.
Artigo em Inglês | MEDLINE | ID: mdl-30612502

RESUMO

Background The molecular mechanisms through which high-demand pacing induce myocardial dysfunction remain unclear. Methods and Results We created atrioventricular block in pigs using dependent right ventricular septal pacing for 6 months. Echocardiography was performed to evaluate dyssynchrony between pacing (n=6) and sham control (n=6) groups. Microarray and enrichment analyses were used to identify differentially expressed genes ( DEG s) in the left ventricular ( LV ) myocardium between pacing and sham control groups. Histopathological and protein changes were also analyzed and an A cell pacing model was also performed. Pacing significantly increased mechanical dyssynchrony. Enrichment analysis using Ingenuity Pathway Analysis and the activation z-score analysis method demonstrated that there were 5 DEG s ( ABCA 1, APOD , CLU , LY 96, and SERPINF 1) in the LV septum (z-score=-0.447) and 5 DEG s ( APOD , CLU , LY 96, MSR 1, and SERPINF 1) in the LV free wall (z-score=-1.000) inhibited the liver X receptor/retinoid X receptor ( LXR / RXR ) pathway, and 4 DEG s ( ACTA 2, MYL 1, PPP 2R3A, and SNAI 2) activated the integrin-linked kinase ( ILK ) pathway in the LV septum (z-score=1.000). The pacing group had a larger cell size, higher degree of myolysis and fibrosis, and increased expression of intracellular lipid, inflammatory cytokines, and apoptotic markers than the sham control group. The causal relationships between pacing and DEG s related to LXR / RXR and ILK pathways, apoptosis, fibrosis, and lipid expression after pacing were confirmed in the cell pacing model. Luciferase reporter assay in the cell pacing model also supported inhibition of the LXR pathway by pacing. Conclusions Right ventricular septal-dependent pacing was associated with persistent LV dyssynchrony-induced cardiomyopathy through inhibition of the LXR / RXR pathway.


Assuntos
Cardiomiopatias/metabolismo , Regulação da Expressão Gênica , Ventrículos do Coração/metabolismo , Receptores X do Fígado/genética , Miócitos Cardíacos/metabolismo , Receptores X Retinoide/genética , Animais , Apoptose , Western Blotting , Cardiomiopatias/diagnóstico , Cardiomiopatias/etiologia , Células Cultivadas , Modelos Animais de Doenças , Ecocardiografia , Eletrocardiografia , Feminino , Citometria de Fluxo , Ventrículos do Coração/patologia , Receptores X do Fígado/metabolismo , Miócitos Cardíacos/patologia , Marca-Passo Artificial/efeitos adversos , RNA/genética , Receptores X Retinoide/metabolismo , Suínos , Porco Miniatura
7.
J Agric Food Chem ; 67(1): 140-147, 2019 Jan 09.
Artigo em Inglês | MEDLINE | ID: mdl-30418775

RESUMO

To investigate the effect of heat treatment on the antitumor activity of lactoferrin in colon cancer cells and colon tumors, the HT-29 (human intestinal epithelial tumor cell) cell line was exposed to lactoferrin and various heat treatments. The impacts on cell proliferation, invasion, and migration were observed in vitro, and nude mice bearing HT29 tumors were administered lactoferrin and underwent various heat treatments in vivo. In the HT29 cell proliferation test using transwell and scratch analyses, lactoferrin (20 mg/mL) without or with heat treatment (50 and 70 °C) significantly inhibited cell proliferation, migration, and invasion (compared with the control, p < 0.05), while lactoferrin with heat treatment (100 °C) did not affect these parameters. In vivo, HT29 tumor weight was significantly reduced in the lactoferrin (without heat treatment and with 50 and 70 °C treatment) groups (1.59 ± 0.20, 1.67 ± 0.25, and 2.41 ± 0.42 g, compared with the control, p < 0.05), and there was no significant difference between the control (3.73 ± 0.33 g) and the 100 °C treatment group (3.58 ± 0.29 g). Moreover, 100 °C heat treatment reduced inhibition of the VEGFR2/VEGFA/PI3K/Akt/Erk1/2 angiogenesis pathway by lactoferrin. In summary, HT29 tumors were effectively suppressed by lactoferrin via inhibition of VEGFR2/VEGFA/PI3K/Akt/Erk1/2 pathway, and heat treatment affected the antitumor activity of lactoferrin in a temperature-dependent manner.


Assuntos
Antineoplásicos/administração & dosagem , Neoplasias do Colo/tratamento farmacológico , Lactoferrina/administração & dosagem , Animais , Antineoplásicos/química , Apoptose/efeitos dos fármacos , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Neoplasias do Colo/genética , Neoplasias do Colo/metabolismo , Neoplasias do Colo/fisiopatologia , Células HT29 , Temperatura Alta , Humanos , Lactoferrina/química , Sistema de Sinalização das MAP Quinases , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Fosfatidilinositol 3-Quinases/genética , Fosfatidilinositol 3-Quinases/metabolismo , Fator A de Crescimento do Endotélio Vascular/genética , Fator A de Crescimento do Endotélio Vascular/metabolismo , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/genética , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/metabolismo , Ensaios Antitumorais Modelo de Xenoenxerto
8.
J Cancer ; 9(23): 4521-4526, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30519358

RESUMO

Neuroblastoma is a lethal tumor of the sympathetic nervous system. 8-Hydroxydeoxyguanine (8-OH-dG) formation is a common seen type of oxidative DNA damage, which could be repaired by human oxoguanine glycosylase 1 (hOGG1). To explore the contributing role of hOGG1 gene single nucleotide polymorphisms (SNPs) in neuroblastoma risk, we performed a case-control study by genotyping three SNPs (rs1052133 G>C, rs159153 T>C, rs293795 A>G) in hOGG1 gene. A total of 512 neuroblastoma cases and 1076 cancer-free controls were enrolled from three medical centers in China. The hOGG1 gene polymorphisms were determined using TaqMan real-time PCR. The results showed that only the rs1052133 G>C polymorphism was associated with neuroblastoma risk [GC vs. GG: adjusted odds ratio (OR)=0.64, 95% confidence interval (CI)=0.51-0.81, P=0.0002; dominant model: adjusted OR=0.71, 95% CI=0.57-0.88, P=0.002]. Moreover, subjects carrying 1, 2, or 1-3 protective genotypes have less opportunity to develop neuroblastoma, in comparison to those without protective genotypes. Stratified analysis revealed that rs1052133 GC/CC carriers were less likely to develop neuroblastoma in subgroups of age >18 months, males, tumor that develops from retroperitoneal, mediastinum and clinical stage I+II+4s. Our results indicate that hOGG1 rs1052133 G>C polymorphism is associated with decreased risk of neuroblastoma. However, the exact biological mechanism awaits further research.

9.
Int J Mol Sci ; 19(12)2018 Dec 18.
Artigo em Inglês | MEDLINE | ID: mdl-30567295

RESUMO

BACKGROUND: Lipid expression is increased in the atrial myocytes of mitral regurgitation (MR) patients. This study aimed to investigate key regulatory genes and mechanisms of atrial lipotoxic myopathy in MR. METHODS: The HL-1 atrial myocytes were subjected to uniaxial cyclic stretching for eight hours. Fatty acid metabolism, lipoprotein signaling, and cholesterol metabolism were analyzed by PCR assay (168 genes). RESULTS: The stretched myocytes had significantly larger cell size and higher lipid expression than non-stretched myocytes (all p < 0.001). Fatty acid metabolism, lipoprotein signaling, and cholesterol metabolism in the myocytes were analyzed by PCR assay (168 genes). In comparison with their counterparts in non-stretched myocytes, seven genes in stretched monocytes (Idi1, Olr1, Nr1h4, Fabp2, Prkag3, Slc27a5, Fabp6) revealed differential upregulation with an altered fold change >1.5. Nine genes in stretched monocytes (Apoa4, Hmgcs2, Apol8, Srebf1, Acsm4, Fabp1, Acox2, Acsl6, Gk) revealed differential downregulation with an altered fold change <0.67. Canonical pathway analysis, using Ingenuity Pathway Analysis software, revealed that the only genes in the "superpathway of cholesterol biosynthesis" were Idi1 (upregulated) and Hmgcs2 (downregulated). The fraction of stretched myocytes expressing Nile red was significantly decreased by RNA interference of Idi1 (p < 0.05) and was significantly decreased by plasmid transfection of Hmgcs2 (p = 0.004). CONCLUSIONS: The Idi1 and Hmgcs2 genes have regulatory roles in atrial lipotoxic myopathy associated with atrial enlargement.


Assuntos
Isomerases de Ligação Dupla Carbono-Carbono/genética , Hidroximetilglutaril-CoA Sintase/genética , Metabolismo dos Lipídeos/genética , Insuficiência da Valva Mitral/genética , Linhagem Celular , Colesterol/genética , Colesterol/metabolismo , Citometria de Fluxo , Regulação da Expressão Gênica/genética , Átrios do Coração/metabolismo , Átrios do Coração/fisiopatologia , Hemiterpenos , Humanos , Lipídeos/genética , Lipoproteínas/genética , Lipoproteínas/metabolismo , Insuficiência da Valva Mitral/metabolismo , Insuficiência da Valva Mitral/fisiopatologia , Miócitos Cardíacos/metabolismo , Miócitos Cardíacos/patologia , Transdução de Sinais
10.
Oncotarget ; 8(39): 65386-65396, 2017 Sep 12.
Artigo em Inglês | MEDLINE | ID: mdl-29029438

RESUMO

Iron is an essential nutrient that facilitates cell proliferation and growth, which plays a pivotal role in modulating the battle for survival between mammalian hosts and their pathogens. Pathogenic bacteria secrete siderophores to acquire iron from the host. However, lipocalin 2 (Lcn2), a siderophore-binding antimicrobial protein, binds to siderophores to prevent bacterial uptake of iron, which is critical for the control of systemic infection with Escherichia coli (E. coli). But few studies focus on the anti-infective response of Lcn2 in the intestines by inhibiting bacterial proliferation based on microbial iron metabolism. In this study, we showed that iron was sequestrated within cells in a piglet model of E. coli K88 infection. Siderophores was produced following E. coli K88 infection and siderophore-related genes expression was upregulated in iron-deficiency environment in vitro. Meanwhile, we found that Lcn2 expression was rapidly and robustly induced in jejunum by E. coli K88 infection and could be stimulated by IL-17 and IL-22. Furthermore, both Lcn2 induced in epithelial cells IPEC-1 and added exogenously as a recombinant protein could inhibit the growth of E. coli. We can conclude that Lcn2 is a crucial component of mucosal immune defense against intestinal infection with E. coli K88.

11.
Artigo em Inglês | MEDLINE | ID: mdl-28760901

RESUMO

Hepcidin, an antimicrobial peptide, was discovered to integrate diverse signals from iron status and an infection threat and orchestrate a series of host-protective responses. Several studies have investigated the antimicrobial role of hepcidin, but the results have been controversial. Here, we aimed to examine the role of hepcidin in bacterial adherence and invasion in vitro We found that porcine hepcidin could decrease the amount of the extracellular pathogen enterotoxigenic Escherichia coli (ETEC) K88 that adhered to cells because it caused the aggregation of the bacteria. However, addition of hepcidin to macrophages infected with the intracellular pathogen Salmonella enterica serovar Typhimurium enhanced the intracellular growth of the pathogen through the degradation of ferroportin, an iron export protein, and then the sequestration of intracellular iron. Intracellular iron was unavailable by use of the iron chelator deferiprone (DFO), which reduced intracellular bacterial growth. These results demonstrate that hepcidin exhibits different functions in extracellular and intracellular bacterial infections, which suggests that different defense strategies should be taken to prevent bacterial infection.


Assuntos
Antibacterianos/farmacologia , Aderência Bacteriana/efeitos dos fármacos , Escherichia coli Enterotoxigênica/efeitos dos fármacos , Infecções por Escherichia coli/tratamento farmacológico , Hepcidinas/farmacologia , Infecções por Salmonella/tratamento farmacológico , Salmonella typhimurium/efeitos dos fármacos , Animais , Células CACO-2 , Proteínas de Transporte de Cátions/metabolismo , Linhagem Celular Tumoral , Deferiprona , Escherichia coli Enterotoxigênica/patogenicidade , Infecções por Escherichia coli/microbiologia , Humanos , Ferro/metabolismo , Macrófagos/microbiologia , Piridonas/metabolismo , Infecções por Salmonella/microbiologia , Salmonella typhimurium/crescimento & desenvolvimento , Salmonella typhimurium/patogenicidade , Suínos
12.
Mediators Inflamm ; 2017: 8570818, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28286378

RESUMO

Iron metabolism in inflammation has been mostly characterized in macrophages exposed to pathogens or inflammatory conditions. The aim of this study is to investigate the cross-regulatory interactions between M1 macrophage polarization and iron metabolism. Firstly, we characterized the transcription of genes related to iron homeostasis in M1 RAW264.7 macrophages stimulated by IFN-γ. The molecular signature of M1 macrophages showed high levels of iron storage (ferritin), a low level of iron export (ferroportin), and changes of iron regulators (hepcidin and transferrin receptors), which favour iron sequestration in the reticuloendothelial system and are benefit for inflammatory disorders. Then, we evaluated the effect of iron on M1 macrophage polarization. Iron significantly reduced mRNA levels of IL-6, IL-1ß, TNF-α, and iNOS produced by IFN-γ-polarized M1 macrophages. Immunofluorescence analysis showed that iron also reduced iNOS production. However, iron did not compromise but enhanced the ability of M1-polarized macrophages to phagocytose FITC-dextran. Moreover, we demonstrated that STAT1 inhibition was required for reduction of iNOS and M1-related cytokines production by the present of iron. Together, these findings indicated that iron decreased polarization of M1 macrophages and inhibited the production of the proinflammatory cytokines. The results expanded our knowledge about the role of iron in macrophage polarization.


Assuntos
Ferro/farmacologia , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Fator de Transcrição STAT1/metabolismo , Animais , Proteínas de Transporte de Cátions/metabolismo , Ferritinas/metabolismo , Interferon gama/farmacologia , Interleucina-1beta/genética , Interleucina-6/genética , Camundongos , Óxido Nítrico Sintase Tipo II/genética , Células RAW 264.7 , RNA Mensageiro/genética , Receptores da Transferrina/metabolismo , Fator de Necrose Tumoral alfa/genética
13.
Huan Jing Ke Xue ; 38(10): 4194-4211, 2017 Oct 08.
Artigo em Chinês | MEDLINE | ID: mdl-29965203

RESUMO

Occurrence of 17 polycyclic aromatic hydrocarbons (PAHs) and 15 persistent organochlorine pesticides (OCPs) were analyzed by gas chromatography coupled to tandem mass spectrometry (GC/MS/MS) during the spring and summer of 2015 in surface water at 14 sampling sites in the mainstream of the Luanhe River to elucidate their seasonal distribution, composition, and potential sources. Results showed that total PAH concentrations in the surface water of the Luanhe River ranged from 33.33 ng·L-1 to 90.65 ng·L-1(52.64 ng·L-1 in average) in spring and 147.68 ng·L-1 to 252.68 ng·L-1 (169.88 ng·L-1 in average) in summer, respectively, and total concentrations of OCPs ranged from <0.08 ng·L-1 to 3.48 ng·L-1 (1.19 ng·L-1 in average) in spring and <0.08 to 5.47 ng·L-1(1.02 ng·L-1 in average) in summer, respectively. Seasonal pollution characteristics of PAHs in surface water during two different seasons showed that total PAH concentrations were much higher in summer than in spring at each sampling site on the Luanhe River. While seasonal pollution characteristics of OCPs showed that total OCP concentrations were much lower in spring than in summer at six sites including Luanxian, Daheiting reservoir dam, Panjiakou Village, Guojiatun, Qujiawan, and Baichengzi, and were higher in spring than in summer at the remaining eight sites. From the perspective of spatial and temporal variations in the composition patterns of PAHs and OCPs in surface water of the Luanhe River, the low and middle ring species of PAHs were dominant in spring and summer and the proportion of PAHs with different rings showed significant differences. Three-ring PAHs dominated the samples with 51.18% and 67.55% (61.39% on average) of total PAHs in spring, and four-ring and two-ring PAHs accounted for 11.78%-33.94% (20.97% on average) and 13.31%-27.12% (16.94% on average), respectively. In the summer, In the summer, naphthalene(Nap) was the main component of total PAHs at each sampling site and PAHs were predominated by two-ring compounds with 77.08% and 90.62% (83.34% on average) of the total, and by three-ring and four-ring compounds with 7.09%-15.22% (12.40% on average) and 2.23%-7.49% (3.97% on average), respectively. Only the residues of hexachlorocyclohexanes(HCHs) and dichlorodiphenyltrichloroethanes(DDTs) were detected in surface water of the Luanhe River at different degrees and HCHs were the dominant compounds. ß-HCH was the main pollutant at most sampling sites with 43.78% and 70.09% (56.25% on average) of total HCHs in the spring and with 53.63% and 64.31% (58.37% on average) in the summer. α-HCH and γ-HCH were abundant at other sampling sites. Source analysis based on the isomer ratios indicated that PAHs in the Luanhe River mainly originated from combustion sources and also likely from petroleum processes. Ratios of OCPs confirmed that historical residues and long-range atmospheric transport were the major sources of HCHs in the study area, and new sources of lindane or γ-HCH existed at Wulongji and Sandaohezi in the spring and Daheting reservoir dam in the summer. Fresh inputs at the Panjiakou reservoir dam, Sandaohezi, Waigoumenzi, Baichengzi, Zhenglanqi, and the Shandianhe reservoir, and environmental residues at Jianggezhuang and Guojiatun were the major sources of DDTs. Concentrations of single PAHs in surface water of the Luanhe River did not exceed the water quality standards of China and other countries except that chrysene(Chr) exceeded the United States national water quality standard. Individual PAHs did not exceed any safety guidelines for aquatic organisms that are exposed to PAHs in water but total concentrations of PAHs exceeded the threshold limits set by the United States Environmental Protection Agency (EPA) and the European Union. This result indicated that consuming fish and other aquatic animals might pose a potential threat to human health due to the bioaccumulation of PAHs in aquatic organisms. The concentrations of OCPs in surface water of the Luanhe River did not exceed any water quality standards or the aquatic life threshold limits for freshwater developed by the EPA. However, the concentrations of α-HCH at Jianggezhuang, Wulongji, Qujiawan, and Baichengziand sites in the spring and at the Panjiakou reservoir dam, Panjiakou village, Qujiawan, and Baichengzi sites in the summer, and concentrations of p,p'-DDD at the Shandianhe reservoir and p,p'-DDE at Jianggezhuang, Sandaohezi, Guojiatun, and Shandianhe reservoir in the spring exceeded the human health ambient water quality criteria limit developed by the EPA. This result suggests that α-HCH, p,p'-DDE, and p,p'-DDD pose a potential health risk to the residents who live near the sampling sites on the Luanhe River.


Assuntos
Hidrocarbonetos Clorados/análise , Praguicidas/análise , Hidrocarbonetos Policíclicos Aromáticos/análise , Rios/química , Estações do Ano , Poluentes Químicos da Água/análise , Animais , China , Monitoramento Ambiental , Cromatografia Gasosa-Espectrometria de Massas , Humanos , Espectrometria de Massas em Tandem
14.
Huan Jing Ke Xue ; 38(3): 964-978, 2017 Mar 08.
Artigo em Chinês | MEDLINE | ID: mdl-29965566

RESUMO

Seasonal variations of 17 polycyclic aromatic hydrocarbons(PAHs) and 15 persistent organochlorine pesticides(OCPs) were analyzed during spring and summer in surface water from 12 sampling sites of Baiyangdian lake to elucidate their seasonal pollution characteristics and sources. The results showed that total concentrations of PAHs in surface water of Baiyangdian lake ranged from 35.38 to 88.06 ng·L-1(46.57 ng·L-1 in average) in spring and 25.64 to 301.41 ng·L-1(76.23 ng·L-1 in average) in summer, respectively, and total concentrations of OCPs ranged from 0.69 to 4.50 ng·L-1(1.77 ng·L-1 in average) in spring and 0.11 to 3.20 ng·L-1(0.90 ng·L-1 in average) in summer, respectively. Seasonal pollution characteristics of PAHs in surface water during two different seasons showed that the total concentration of PAHs was much higher in summer than in spring at three sampling sites of Baiyangdian lake including Qiantang, Guancheng and Anxin bridge, and the level of PAHs in spring was much higher than that in summer at the other nine sites. While seasonal pollution characteristics of OCPs showed that the total concentration of OCPs was much higher in summer than in spring in samples of Guancheng sites, and the level of OCPs in spring was slightly higher than that in summer in the rest of samples. From the perspective of spatial and temporal variations in the composition patterns of PAHs and OCPs in surface water of Baiyangdian lake, PAHs were predominated by 3-ring PAHs with 45.92% and 61.36%(52.60% in average) of total PAHs in spring, while in summer Nap was the chief component of 2-ring PAHs in three sampling sites including Qiantang, Guancheng and Anxin bridge with 84.91%, 91.04% and 78.10% of the total, respectively, and 3-ring PAHs were primary pollutants in the other nine sites with 37.14% to 53.90%(48.94% in average) of the total. Residues of HCHs and DDTs were detected in different degrees and ß-HCH was the main pollutant at all sampling sites with 29.94% and 100%(59.87% in average) of total HCHs in spring, while ß-HCH was the predominant compounds at five sampling sites with 57.55% and 80.23%(61.98% in average) in summer and α-HCHs and δ-HCH were abundant at other sampling sites. Source analysis based on isomer ratios indicated that the main source of PAHs in Baiyangdian lake originated from combustion sources, and additional sources for several sampling sites could be petroleum processes. Ratios of HCH isomers and DDT congeners confirmed that environmental historical residues and long range atmospheric transport were the major sources of OCPs contamination in the study area. Compared with the water quality standard of China and other countries or organizations, concentrations of PAHs and OCPs in surface water of Baiyangdian lake did not exceed the target values of the standard. However, the concentrations of α-HCH and p,p'-DDD in Anxin bridge and Quantou sites exceeded the limit of human health ambient water quality criteria developed by EPA, which reflected that α-HCH and p,p'-DDD would have a potential hazards risk to the residents from Baiyangdian district.

15.
Food Funct ; 5(12): 3224-32, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25317925

RESUMO

Antrodia camphorata is a fungus native to Taiwan, and it is considered a precious medicinal agent. We analyzed triterpenoids, polysaccharides and 1,3-ß-D-glucan, three major effective components in A. camphorata extracts (ACE). ACE exhibited a selective cytotoxic effect on BxPC-3 human pancreatic cancer cells. ACE markedly inhibited the migration ability of BxPC-3 cells. Treatment of BxPC-3 cells with ACE resulted in the increase of cells in the sub-G1 phase and G2/M phase arrest. Apoptosis was confirmed by validating phosphatidylserine externalization, the observation of characteristic chromatin condensation, and nuclear DNA fragmentation. ACE induced apoptosis in BxPC-3 cells through a mitochondria-dependent pathway by triggering an appropriate balance of bax/bcl-2, cytochrome c release, activation of caspase-9 and -3, and poly(ADP-ribose) polymerase cleavage. ACE shows great therapeutic potential due to its cytotoxic effects against BxPC-3 cells which include inhibiting cell migration and inducing mitochondria-mediated apoptosis.


Assuntos
Antineoplásicos Fitogênicos/farmacologia , Antrodia/química , Carpóforos/química , Glucanos/farmacologia , Neoplasias Pancreáticas/fisiopatologia , Polissacarídeos/farmacologia , Triterpenos/farmacologia , Antineoplásicos Fitogênicos/química , Apoptose/efeitos dos fármacos , Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Citocromos c/metabolismo , Fragmentação do DNA/efeitos dos fármacos , Glucanos/química , Humanos , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Neoplasias Pancreáticas/tratamento farmacológico , Polissacarídeos/química , Triterpenos/química
16.
Can J Microbiol ; 60(3): 113-20, 2014 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-24588384

RESUMO

Antimicrobial peptides (AMPs) are part of the innate immune system of complex multicellular organisms. Despite the fact that AMPs show great potential as a novel class of antibiotics, the lack of a cost-effective means for their mass production limits both basic research and clinical use. In this work, we describe a novel expression system for the production of antimicrobial peptides in Escherichia coli by combining ΔI-CM mini-intein with the self-assembling amphipathic peptide 18A to drive the formation of active aggregates. Two AMPs, human ß-defensin 2 and LL-37, were fused to the self-cleaving tag and expressed as active protein aggregates. The active aggregates were recovered by centrifugation and the intact antimicrobial peptides were released into solution by an intein-mediated cleavage reaction in cleaving buffer (phosphate-buffered saline supplemented with 40 mmol/L Bis-Tris, 2 mmol/L EDTA, pH 6.2). The peptides were further purified by cation-exchange chromatography. Peptides yields of 0.82 ± 0.24 and 0.59 ± 0.11 mg/L were achieved for human ß-defensin 2 and LL-37, respectively, with demonstrated antimicrobial activity. Using our expression system, intact antimicrobial peptides were recovered by simple centrifugation from active protein aggregates after the intein-mediated cleavage reaction. Thus, we provide an economical and efficient way to produce intact antimicrobial peptides in E. coli.


Assuntos
Anti-Infecciosos/metabolismo , Peptídeos Catiônicos Antimicrobianos/metabolismo , Escherichia coli/metabolismo , Sequência de Aminoácidos , Anti-Infecciosos/química , Peptídeos Catiônicos Antimicrobianos/química , Peptídeos Catiônicos Antimicrobianos/genética , Peptídeos Catiônicos Antimicrobianos/farmacologia , Candida albicans/efeitos dos fármacos , Escherichia coli/química , Escherichia coli/genética , Escherichia coli K12/efeitos dos fármacos , Regulação Bacteriana da Expressão Gênica , Humanos , Inteínas , Peptídeos/química , Peptídeos/genética , Peptídeos/metabolismo , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Trometamina/análogos & derivados , beta-Defensinas/química , beta-Defensinas/genética , beta-Defensinas/metabolismo , beta-Defensinas/farmacologia
17.
Appl Microbiol Biotechnol ; 98(8): 3651-58, 2014 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-24121930

RESUMO

Small ubiquitin-related modifier (SUMO) technology has been widely used in Escherichia coli expression systems to produce antimicrobial peptides. However, E. coli is a pathogenic bacterium that produces endotoxins and can secrete proteins into the periplasm, forming inclusion bodies. In our work, cathelicidin-BF (CBF), an antimicrobial peptide purified from Bungarus fasciatus venom, was produced in a Bacillus subtilis expression system using SUMO technology. The chimeric genes his-SUMO-CBF and his-SUMO protease 1 were ligated into vector pHT43 and expressed in B. subtilis WB800N. Approximately 22 mg of recombinant fusion protein SUMO-CBF and 1 mg of SUMO protease 1 were purified per liter of culture supernatant. Purified SUMO protease 1 was highly active and cleaved his-SUMO-CBF with an enzyme-to-substrate ratio of 1:40. Following cleavage, recombinant CBF was further purified by affinity and cation exchange chromatography. Peptide yields of ~3 mg/l endotoxin-free CBF were achieved, and the peptide demonstrated antimicrobial activity. This is the first report of the production of an endotoxin-free antimicrobial peptide, CBF, by recombinant DNA technology, as well as the first time purified SUMO protease 1 with high activity has been produced from B. subtilis. This work has expanded the application of SUMO fusion technology and may represent a safe and efficient way to generate peptides and proteins in B. subtilis.


Assuntos
Bacillus subtilis/metabolismo , Catelicidinas/biossíntese , Catelicidinas/genética , Expressão Gênica , Engenharia Metabólica/métodos , Proteína SUMO-1/biossíntese , Proteína SUMO-1/genética , Animais , Bacillus subtilis/genética , Cromatografia de Afinidade , Hidrólise , Proteínas Recombinantes de Fusão/biossíntese , Proteínas Recombinantes de Fusão/genética
18.
Gene ; 532(2): 246-52, 2013 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-24055422

RESUMO

Skeletal muscle fiber type composition is one of the important factors influencing muscle growth and meat quality. As a member of the myogenic transcription factors, myogenin (MyoG) is required for embryonic myoblast differentiation, but the expression of MyoG continues in mature muscle tissue of adult animals, especially in oxidative metabolic muscle, which suggests that MyoG may play a more extended role. Therefore, using MyoG gene transfer mice and C2C12 myoblasts as in vivo and in vitro models, respectively, we elected to study the role of MyoG in muscle fiber types and oxidative metabolism by using overexpression and siRNA suppression strategies. The overexpression of MyoG by DNA electroporation in mouse gastrocnemius muscle had no significant effect on fiber type composition but upregulated the mRNA expression (P<0.01) and enzyme activity (P<0.05) of oxidative succinic dehydrogenase (SDH). In addition, downregulation of the activity of the glycolytic enzymes lactate dehydrogenase (LDH, P<0.05) and pyruvate kinase (PK, P<0.05) was observed in MyoG gene transfer mice. In vitro experiments verified the results obtained in mice. Stable MyoG-transfected differentiating C2C12 cells showed higher mRNA expression levels of myosin heavy chain (MyHC) isoform IIX (P<0.01) and SDH (P<0.05), while the LDH mRNA was attenuated. The enzyme activities of SDH (P<0.01) and LDH (P<0.05) were similarly altered at the mRNA level. When MyoG was knocked down in C2C12 cells, MyHC IIX expression (P<0.05) was decreased, but the mRNA level (P<0.05) and the enzyme activity (P<0.05) of SDH were increased. Downregulating MyoG also increased the activity of the glycolytic enzymes PK (P<0.05) and hexokinase (HK, P<0.05). Based on those results, we concluded that MyoG barely changes the MyHC isoforms, except MyHC IIX, in differentiating myoblasts but probably influences the shift from glycolytic metabolism towards oxidative metabolism both in vivo and in vitro. These results contribute to further understand the role of MyoG in skeletal muscle energy metabolism and also help to explore the key genes that regulate meat quality.


Assuntos
Fibras Musculares Esqueléticas/enzimologia , Mioblastos/enzimologia , Miogenina/genética , Animais , Diferenciação Celular , Linhagem Celular , Qualidade dos Alimentos , Expressão Gênica , Regulação Enzimológica da Expressão Gênica , Técnicas de Silenciamento de Genes , Técnicas de Transferência de Genes , Glicólise , L-Lactato Desidrogenase/metabolismo , Malato Desidrogenase/metabolismo , Masculino , Carne/normas , Camundongos , Camundongos Endogâmicos ICR , Mitocôndrias Musculares/metabolismo , Fibras Musculares Esqueléticas/metabolismo , Músculo Esquelético/citologia , Músculo Esquelético/enzimologia , Mioblastos/metabolismo , Mioblastos/fisiologia , Miogenina/biossíntese , Cadeias Pesadas de Miosina/genética , Cadeias Pesadas de Miosina/metabolismo , RNA Interferente Pequeno/genética , Succinato Desidrogenase/metabolismo , Sus scrofa
19.
Biomed Res Int ; 2013: 754319, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23936842

RESUMO

OG2 is a modified antimicrobial peptide, that is, derived from the frog peptide Palustrin-OG1. It has high antimicrobial activity and low cytotoxicity, and it is therefore promising as a therapeutic agent. Both prokaryotic (Escherichia coli) and eukaryotic (Pichia pastoris) production host systems were used to produce OG2 in our previous study; however, it was difficult to achieve high expression yields and efficient purification. In this study, we achieved high-yield OG2 expression using the intein fusion system. The optimized OG2 gene was cloned into the pTWIN1 vector to generate pTWIN-OG2-intein2 (C-terminal fusion vector) and pTWIN-intein1-OG2 (N-terminal fusion vector). Nearly 70% of the expressed OG2-intein2 was soluble after the IPTG concentration and induction temperature were decreased, whereas only 42% of the expressed of intein1-OG2 was soluble. Up to 75 mg of OG2-intein2 was obtained from a 1l culture, and 85% of the protein was cleaved by 100 mM DTT. Intein1-OG2 was less amenable to cleavage due to the inhibition of cleavage by the N-terminal amino acid of OG2. The purified OG2 exhibited strong antimicrobial activity against E. coli K88. The intein system is the best currently available system for the cost-effective production of OG2.


Assuntos
Peptídeos Catiônicos Antimicrobianos/biossíntese , Peptídeos Catiônicos Antimicrobianos/isolamento & purificação , Inteínas/genética , Proteínas Recombinantes de Fusão/biossíntese , Animais , Anti-Infecciosos , Peptídeos Catiônicos Antimicrobianos/genética , Clonagem Molecular , Escherichia coli/genética , Vetores Genéticos , Humanos , Pichia/genética , Proteínas Recombinantes de Fusão/genética
20.
J Chromatogr A ; 1307: 41-8, 2013 Sep 13.
Artigo em Inglês | MEDLINE | ID: mdl-23921262

RESUMO

A simple and cost-effective pre-treatment procedure was developed for six resorcylic acid lactones (RALs) in feed using dispersive solid phase extraction (dSPE) with multi-walled carbon nanotubes (MWCNTs). The sample was analysed after purification by ultra-high performance liquid chromatography-negative electrospray ionisation tandem mass spectrometry (UHPLC-ESI-MS/MS). After extraction with acetonitrile/water (80:20, v/v) and dilution with water, a dSPE procedure was carried out with MWCNTs. The pH value of the extract, the extraction time for MWCNTs, the type and amount of MWCNTs and the type of eluent were optimised to increase the sample throughput and the sensitivity. The samples were quantified using the internal standard zearalenone-D6. The absolute recoveries of the target compounds from feed samples were most efficient when using 100mg of MWCNTs with an outer diameter of less than 8nm and a length of 10-30µm, and ethyl acetate was shown to be the most suitable solvent for desorbing the target compounds from the MWCNTs. The mean recoveries from fortified swine mixed feed samples ranged from 95.3% to 107.2% and had relative standard deviations lower than 10%; the limits of detection and quantification for RALs were in the ranges of 0.20-0.29µg/kg and 0.54-0.78µg/kg, respectively.


Assuntos
Cromatografia Líquida/métodos , Estrogênios não Esteroides/análise , Extração em Fase Sólida/métodos , Espectrometria de Massas em Tandem/métodos , Zearalenona/análogos & derivados , Zearalenona/análise , Zeranol/análogos & derivados , Acetatos/química , Acetonitrilos/química , Ração Animal/análise , Estrogênios não Esteroides/química , Estrogênios não Esteroides/isolamento & purificação , Concentração de Íons de Hidrogênio , Limite de Detecção , Modelos Lineares , Nanotubos de Carbono/química , Reprodutibilidade dos Testes , Zearalenona/química , Zearalenona/isolamento & purificação , Zeranol/análise , Zeranol/química , Zeranol/isolamento & purificação
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