RESUMO
BACKGROUND: Malonaldehyde, the primary by-product of lipid peroxidation in food, modifies the structural and functional properties of proteins by cross-linking. The aim of this study was to investigate the effect of malonaldehyde on the allergenicity of shrimp tropomyosin. RESULTS: RBL-2H3 cells, a model of type I allergic reactions, were sensitised with sera from patients allergic to shrimp, and were stimulated with native and cross-linked tropomyosin. Release of inflammatory mediators such as ß-hexosaminidase, histamine, tryptase, cysteinyl leukotriene, and prostaglandin D2 was clearly suppressed in a manner that depended on the extent of tropomyosin cross-linking. Release of interleukin-4 (IL-4) and IL-13 was similarly decreased. Notably, cells sensitised with one patient's serum released IL-4 at comparable levels in response to native and cross-linked tropomyosin. CONCLUSION: Cross-linking strongly modulates the ability of shrimp tropomyosin to induce release of inflammatory cytokines and mediators from activated RBL-2H3 cells. © 2016 Society of Chemical Industry.
Assuntos
Alérgenos/efeitos dos fármacos , Reagentes de Ligações Cruzadas/farmacologia , Citocinas/metabolismo , Mediadores da Inflamação/metabolismo , Malondialdeído/farmacologia , Tropomiosina/farmacologia , Animais , Linhagem Celular , Citocinas/imunologia , Hipersensibilidade Alimentar/sangue , Hipersensibilidade Alimentar/imunologia , Humanos , Mediadores da Inflamação/imunologia , Penaeidae/química , Penaeidae/imunologia , Ratos , Tropomiosina/imunologiaRESUMO
A twelve-week feeding experiment was undertaken to explore the impact of substituting dietary fish meal (FM) and fish oil (FO) with complex protein (CP) and canola oil (CO) in the diet of triploid rainbow trout on the quality of their fillets. The control diet (F100) contained FM (60%) and FO (18.6%) as the main protein and lipid sources. Based on this, 50% and 100% of FM and FO were substituted by CP and CO and they were named as F50 and F0, respectively. The results showed that there were no significant differences in the specific growth rates, condition factors, gutted yields, fillet yields and yellowness values as the substitution levels increased (p > 0.05). The F50 treatment obtained the highest values of fillet springiness and chewiness, improved the umami and bitter taste of the fillets by increasing the contents of inosine-5'-monophosphate and histidine, and increased lipid, protein, C18: 1n-9 and C18: 2n-6 contents (p < 0.05). The F0 treatment obtained the highest values of fillet hardness and pH, attenuated the sweet taste of the fillets by decreasing the content of glycine, and decreased the contents of EPA and DHA (p < 0.05). Both F50 and F0 treatments could increase the redness value, decrease the lightness and hue values of fillets, and increase the odor intensity, resulting in the typical fillet odors of green, fatty, orange and fishy (p < 0.05). In general, 50% and 100% of FM and FO substitution did not affect the growth of trout, but it did affect quality. Compared to the F100 treatment, the fillet quality of the F0 treatment was similar to the F50 treatment and could improve the appearance and odor intensity of the fillets. However, the difference was that the F50 treatment increased the springiness, umami, bitterness and lipid nutritional value of the fillets, but the F0 treatment increased the hardness, decreased the sweetness, and decreased the lipid, EPA and DHA contents of the fillets.
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OBJECTIVE: To discuss the regulating mechanism of iron regulatory protein-2 (IRP2) in the iron metabolism of lung cancer. METHODS: The cultured A549 cells were divided into 3 groups: liposome group (including liposomes 20 mg/L), random oligonucleotide group (SCODN group) and antisense oligonucleotide group (ASODN group). And the liposome-mediated transfection was employed with the liposome and SCODN groups as controls. Reverse transcription-polymerase chain reaction (RT-PCR) and Western blot were used to examine the mRNA and protein expressions of iron metabolism-related transferring (Tf), transferrin receptor (TfR) and ferritin (Fn) genes, etc. RESULTS: After a 48-hour transfection, the mRNA expression of Tf had no statistically significant difference among three groups (F = 2.18, P = 0.078); the mRNA expression of TfR in the ASODN group was significantly lower than that in the liposome and SCODN groups (P < 0.05). The expression of Fn mRNA in the ASODN group (0.56 ± 0.06) was higher than that in the liposome (0.36 ± 0.05) and SCODN groups (0.39 ± 0.03) (P < 0.05). After a 48-hour transfection, the IRP2 protein expression of the ASODN group was significantly lower than those of the liposome and SCODN groups (P < 0.05). The Tf protein expression was not statistically different in three groups (F = 2.67, P = 0.088). The TfR protein expression of the ASODN group was lower than those of the liposome and SCODN groups (P < 0.05). And the Fn protein expression of the ASODN group was higher than those of the liposome and SCODN groups (P < 0.05). CONCLUSION: IRP2 may affect the expressions of TfR and Fn in lung adenocarcinoma A549 cells by changing the amount of protein and regulating the iron metabolism.
Assuntos
Adenocarcinoma/metabolismo , Proteína 2 Reguladora do Ferro/metabolismo , Ferro/metabolismo , Neoplasias Pulmonares/metabolismo , Adenocarcinoma de Pulmão , Linhagem Celular Tumoral , Ferritinas/metabolismo , Humanos , Proteína 2 Reguladora do Ferro/genética , Lipossomos/administração & dosagem , Oligodesoxirribonucleotídeos Antissenso/genética , Receptores da Transferrina/metabolismoRESUMO
OBJECTIVE: To explore the relationship between the sputum levels of high mobility group protein B1 (HMGB1) and airway inflammation in bronchial asthma and chronic obstructive pulmonary disease (COPD) patients. METHODS: A total of 57 patients with persistent asthma [per Global Initiative for Asthma (GINA) guidelines], 30 patients with stable COPD [stratified by Global Initiative for COPD (GOLD) status] and 20 control subjects were recruited. After completing an asthma control questionnaire, spirometry was performed before sputum induction. The ratio of forced expiratory volume in the first second (FEV(1))/predictive value (FEV(1)%Pre) and neutrophil differential count in induced sputum were recorded. The concentrations of HMGB1 in the supernatant of sputum were measured by ELISA (enzyme-linked immunosorbent assay). RESULTS: The sputum concentrations of HMGB1 in the asthmatics and COPD patients were significantly higher than those of the control subjects [(291 ± 55) and (511 ± 39) vs (61 ± 5) ng/L, all P < 0.01]. And they were significantly negatively correlated with FEV(1)%Pre in all subjects. The levels of HMGB1 in induced sputum of COPD patients were significantly higher than those of asthmatics and healthy controls (P < 0.01). No significant difference existed in the levels of HMGB1 between patients with eosinophilic and noneosinophilic asthma [(290 ± 55) vs (292 ± 54) ng/L, P > 0.05]. The HMGB1 levels with COPD stage II and stage III were significantly higher than those with stage I [(526 ± 29) and (541 ± 29) vs (471 ± 18) ng/L]. The differences of sputum neutrophil percentage were statistically significant in mild, moderate and severe asthma [(27 ± 2)%, (36 ± 4)%, (49 ± 4)%]. And the sputum levels of HMGB1 were significantly higher in the patients with moderate and severe asthma [(312 ± 14) vs (347 ± 11) ng/L]. And the levels of HMGB1 in asthmatic and COPD patients were positively correlated with neutrophil percentage. According to the multivariate analysis, neutrophil percentage and FEV(1)%Pre were independent predictors of sputum HMGB1, but not smoking, age, gender and eosinophilic percentage. CONCLUSION: HMGB1 may contribute to airway inflammation through its higher expression in bronchial asthma and COPD patients.
Assuntos
Asma/metabolismo , Proteína HMGB1/metabolismo , Doença Pulmonar Obstrutiva Crônica/metabolismo , Adulto , Estudos de Casos e Controles , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Escarro/metabolismoRESUMO
OBJECTIVE: To explore the relationship between HMGB1 (high mobility group box-1) protein and receptor for advanced glycosylation end products (RAGE) and the nosogenesis and severity of bronchial asthma. METHODS: Based on the criteria, the asthma group included 64 acute-onset asthma patients while the control group had 20 healthy cases. The asthma group received a 4-week combination inhalation therapy of budesonide and formoterol. Lung functions and induced sputum examinations were conducted before and after treatment. The percentage of a second forced expiratory volume in the predicted value (FEV(1)%) was recorded. A differential count of neutrophilic leukocyte in reduced sputum was performed. And the sputum levels of HMGB1 and RAGE were detected by ELISA (enzyme linked immunosorbent assay). RESULTS: Prior to treatment, the neutrophilic leukocyte percentage and the levels of HMGB1 and RAGE were all higher than those of control group (P < 0.01). The induced sputum of severe asthma patients had higher levels of neutrophilic leukocyte percentage and HMGB1 than those of mild cases (P < 0.01). But the level of RAGE showed no statistical significance among mild, moderate or severe asthma cases (P > 0.05). The post-treatment levels of neutrophilic leukocyte percentage, HMGB1 and RAGE were lower as compared with the pre-treatment ones (P < 0.01). These three parameters in uncontrolled cases were higher than those in completely controlled cases (P < 0.05); in asthma group, both HMGB1 and RAGE had a negative correlation with FEV(1)% (r = -0.830, r = -0.632, P < 0.01); in induced sputum, both HMGB1 and RAGE had a positive correlation with FEV(1)% (r = 0.820, r = 0.623, P < 0.01). The levels of HMGB1 and RAGE were positively correlated (r = 0.929, P < 0.01). CONCLUSION: Both HMGB1 and RAGE participate in the inflammatory process of asthmatic airway. HMGB1 is correlated with the severity of asthma. And the levels of HMGB1 and RAGE in induced sputum may be employed as reference indices for the observation of therapeutic effects.
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Asma/metabolismo , Proteína HMGB1/metabolismo , Receptores Imunológicos/metabolismo , Escarro/metabolismo , Adulto , Estudos de Casos e Controles , Feminino , Produtos Finais de Glicação Avançada/metabolismo , Humanos , Masculino , Pessoa de Meia-Idade , Receptor para Produtos Finais de Glicação AvançadaRESUMO
High glucose metabolism is recognized as one of the hallmarks of cancer and increased expression levels of several key factors involved in glucose metabolism have been reported in non-small cell lung cancer (NSCLC). Previous studies showed that microRNA (miR)-218 is reduced in NSCLC, but its function in glucose metabolism in NSCLC is not fully understood. The present study aimed to investigate the effect of miR-218 on glucose metabolism in NSCLC cell lines and the underlying molecular mechanism. The present results suggested that miR-218 reduced glucose consumption, the mechanism of glycolysis and activity in the pentose phosphate pathway. In addition, glucose transporter 1 (GLUT1) was identified to be a direct target of miR-218, while overexpression of GLUT1 did not abolish the effect of miR-218 on glucose metabolism. The present results indicated that phosphorylation of NF-κB p65 was significantly decreased by miR-218 in NSCLC cells and that activation of NF-κB led to the inhibition of miR-218 regulation of glucose metabolism. In conclusion, the present results suggested that miR-218 downregulated glucose metabolism in NSCLC not only by directly targeting GLUT1, but also via the NF-κB signaling pathway.
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The aim of the present study was to assess pH-induced changes in conformational structures and potential allergenicity of tropomyosin from short-neck clams. As defined with circular dichroism (CD), an unfolded structure was found at pH values ranging from 2.0 to 5.0, followed by the loss of secondary structure at pH of 1.0. Correspondingly, surface hydrophobicity was reduced by 97.7% when pH was reduced from 7.0 to 1.0. Further indirect ELISA and dot-blot results of pH shifted tropomyosin showed that potential allergenicity correlated well with structural changes, as well as with SGF digestibility. Allergenicity decreased significantly with unfolding of the protein and was stable when surface hydrophobicity recovered back to neutral conditions. These results showed that conformational changes in tropomyosin induced by pH shifting significantly influenced the allergenicity of tropomyosin, and that the resulting changes occurred predominately in the acidic pH range.
Assuntos
Bivalves/imunologia , Imunoglobulina E/química , Tropomiosina/química , Alérgenos/química , Alérgenos/imunologia , Animais , Dicroísmo Circular , Eletroforese em Gel de Poliacrilamida , Ensaio de Imunoadsorção Enzimática , Concentração de Íons de Hidrogênio , Interações Hidrofóbicas e Hidrofílicas , Imunoglobulina G/química , Ligação Proteica , Conformação Proteica , Estrutura Secundária de Proteína , Frutos do Mar/análise , Tropomiosina/imunologiaRESUMO
The aim of this work was to determine the effect of malondialdehyde (MDA) treatment of shrimp tropomyosin (TM) with respect to IgE binding capacity and conformational structure. Following treatment with MDA, changes in TM molecular weight were characterized by SDS-PAGE and TM cross-linking was observed, then the aggregates were recognized by IgG/IgE in immunoblot analysis. Meanwhile, TM allergenicity slowly decreased following MDA treatment. These data show a correlation between the loss of TM structure and allergenic potential. TM secondary structure became more disordered following treatment with increasing concentrations of MDA. Moreover, the enhancement of surface hydrophobicity was also in accordance with the effect caused by MDA. Additionally, MDA treatment resulted in an increase in carbonyl content and a decrease in free amine groups and available lysine. The results suggest that MDA-induced conformational changes in TM can significantly influence the antigenicity and allergenicity of TM.
Assuntos
Alérgenos/química , Imunoglobulina E/química , Malondialdeído/química , Penaeidae/química , Tropomiosina/química , Alérgenos/imunologia , Alérgenos/metabolismo , Animais , Eletroforese em Gel de Poliacrilamida/métodos , Imunoglobulina E/imunologia , Imunoglobulina E/metabolismo , Malondialdeído/farmacologia , Penaeidae/imunologia , Ligação Proteica , Conformação Proteica , Frutos do Mar/análise , Tropomiosina/imunologia , Tropomiosina/metabolismoRESUMO
This study aimed to investigate the effects of Ser/Cys polymorphism in hOGG1 gene, Arg/Pro polymorphism in p53 gene, smoking and their interactions on the development of lung cancer. Ser/Cys polymorphism in hOGG1 and Arg/Pro polymorphism in p53 among 124 patients with lung cancer and 128 normal people were detected using PCR-RFLP. At the same time, smoking status was investigated between the two groups. Logistic regression was used to estimate the effects of Ser/Cys polymorphism and Arg/Pro polymorphisms, smoking and their interactions on the development of lung cancer. ORs (95% CI) of smoking, hOGG1 Cys/Cys and p53 Pro/ Pro genotypes were 2.34 (1.41-3.88), 2.12 (1.03-4.39), and 2.12 (1.15-3.94), respectively. The interaction model of smoking and Cys/Cys was super-multiplicative or multiplicative, and the OR (95% CI) for their interaction item was 1.67 (0.36 -7.78). The interaction model of smoking and Pro/Pro was super-multiplicative with an OR (95%CI) of their interaction item of 5.03 (1.26-20.1). The interaction model of Pro/Pro and Cys/Cys was multiplicative and the OR (95%CI) of their interaction item was 0.99 (0.19-5.28). Smoking, hOGG1 Cys/Cys, p53 Pro/Pro and their interactions may be the important factors leading to the development of lung cancer.