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1.
J Ethnopharmacol ; 251: 112554, 2020 Jan 07.
Artigo em Inglês | MEDLINE | ID: mdl-31923541

RESUMO

ETHNOPHARMACOLOGICAL RELEVANCE: Ulcerative colitis (UC) is tightly associated with inflammation response and oxidative stress. As a folk medicine applied in treatment of diarrhea, Bruguiera gymnorrhiza also possesses anti-inflammatory and anti-oxidative activities, which indicated that B. gymnorrhiza may exert anti-colitis effect. AIM OF THE STUDY: To investigate effect and mechanism of B. gymnorrhiza on experimental UC. MATERIALS AND METHODS: Aqueous extract of B. gymnorrhiza leaves (ABL) was used for investigation in the present study. Murine UC was established through access to 3% dextran sulfate sodium (DSS) for 7 days. Meanwhile, mice accepted treatment with ABL (25, 50, 100 mg/kg) or sulfasalazine (200 mg/kg) once daily. On the last day, disease activity index (DAI) including body weight loss, fecal character and degree of bloody diarrhea was evaluated, colon segments were obtained for length measurement and further analysis and feces were collected for intestinal microbiota analysis. RESULTS: ABL ameliorated DAI scores, colon length shortening and histopathological damage in DSS-induced colitis mice obviously. SOD activity, levels of MDA and GSH altered by colitis were restored remarkably after ABL treatment. ABL inhibited increases in levels of colonic COX-2, iNOS, TNF-α, IL-6, IL-1ß, IL-4, IL-10 and IL-11 in colitis mice. Moreover, ABL prominently suppressed NF-κB p65 and IκB phosphorylation and down-regulated mRNA levels of COX-2, iNOS, TNF-α, IL-6 and IL-1ß elevated by colitis. As shown in microbiota analysis, ABL modulated composition of intestinal microbiota of colitis mice. CONCLUSION: ABL exhibited protective effect against DSS-induced ulcerative colitis through suppressing NF-κB activation and modulating intestinal microbiota.

2.
Ying Yong Sheng Tai Xue Bao ; 30(12): 4333-4343, 2019 Dec.
Artigo em Chinês | MEDLINE | ID: mdl-31840480

RESUMO

The regulation on carbon acquisition and water loss plays a critical role in plant growth and survival. Stomata are important portals for plants to control the exchanges of carbon and water between leaves and the atmosphere. Therefore, understanding stomatal control mechanisms and modelling stomatal conductivity are indispensable to accurately simulate carbon and water cycling in terrestrial ecosystems. As global climate change is accelerating in recent years, drought events have become more and more frequent and thus profoundly affect the survival, growth and distribution of plants. In order to deeply understand the underlying mechanism of carbon-water coupling of plants and predict the dynamics of plants and communities under global changes, it is crucial to explore responses of stomatal regulation of plants to drought stress. In this review, we synthesized recent research progress on mechanisms and modeling of plant stomatal regulation under drought stress. First, this review described the active and passive regulation of plant stomatal control in response to drought stress, and discussed the evolution of plant stomatal regulation, including the passive hydraulic regulation of ferns and lycophytes, the active regulation of angiosperms, and the dual-control mechanism of gymnosperms that was proposed as an important transitional type during evolution from ferns to angiosperms. Then, we analyzed the relationship between stomatal and hydraulic regulations, and discussed the debates on the decoupling of plant water potential from stomatal conductivity. The application of stomatal-conductivity optimization models was introduced based on the water use efficiency hypothesis and the maximum carbon gain hypothesis. The model based on the latter had a greater potential of prediction and practical application. Finally, we proposed two issues that should be urgently addressed: 1) to scale up the research of plant stomatal regulation from leaf or individual to ecosystem or even larger scales so as to improve the mechanistic models of carbon and water cycling in terrestrial ecosystems; and 2) to quantify the hydroactive feedback processes of plant stomatal regulation so as to modify current hydraulics models of plant stomatal function.


Assuntos
Secas , Gleiquênias , Ecossistema , Folhas de Planta , Estômatos de Plantas , Transpiração Vegetal , Água
3.
Food Chem Toxicol ; 123: 349-362, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30423402

RESUMO

Acetaminophen (APAP) overdose-induced hepatotoxicity is tightly associated with oxidative stress. Tetrahydrocurcumin (THC) and octahydrocurcumin (OHC), the primary and final hydrogenated metabolites of curcumin (CUR), possess stronger antioxidant activity in vitro. The present study was performed to investigate the potential and mechanism of OHC and THC against APAP-induced hepatotoxicity in parallel to CUR. Our results showed that OHC and THC dose-dependently enhanced liver function (ALT and AST levels) and alleviated histopathological deterioration. Besides, OHC and THC significantly restored the hepatic antioxidant status by miring level of MDA and ROS, and elevated levels of GSH, SOD, CAT and T-AOC. In addition, OHC and THC markedly suppressed the activity and expressions of CYP2E1, and bound to the active sites of CYP2E1. Moreover, OHC and THC activated the Keap1-Nrf2 pathway and enormously enhanced the translational activation of Nrf2-targeted gene (GCLC, GCLM, NQO1 and HO-1) against oxidative stress, via inhibiting the expression of Keap1 and blocking the interaction between Keap1 and Nrf2. Particularly, OHC and THC exerted superior hepato-protective and antioxidant activities to CUR. In conclusion, OHC and THC possess favorable hepato-protective effect through restoring antioxidant status, inhibiting CYP2E1 and activating Keap1-Nrf2 pathway, which might represent promising antioxidants for the treatment of APAP-induced hepatotoxicity.


Assuntos
Acetaminofen/efeitos adversos , Doença Hepática Induzida por Substâncias e Drogas/prevenção & controle , Curcumina/administração & dosagem , Citocromo P-450 CYP2E1/metabolismo , Proteína 1 Associada a ECH Semelhante a Kelch/metabolismo , Fator 2 Relacionado a NF-E2/metabolismo , Substâncias Protetoras/administração & dosagem , Animais , Doença Hepática Induzida por Substâncias e Drogas/etiologia , Doença Hepática Induzida por Substâncias e Drogas/genética , Doença Hepática Induzida por Substâncias e Drogas/metabolismo , Curcumina/análogos & derivados , Curcumina/química , Curcumina/metabolismo , Citocromo P-450 CYP2E1/genética , Humanos , Hidrogenação , Proteína 1 Associada a ECH Semelhante a Kelch/genética , Fígado/efeitos dos fármacos , Fígado/metabolismo , Masculino , Camundongos , Fator 2 Relacionado a NF-E2/genética , Estresse Oxidativo/efeitos dos fármacos , Substâncias Protetoras/química , Substâncias Protetoras/metabolismo , Transdução de Sinais/efeitos dos fármacos
4.
Front Pharmacol ; 9: 1181, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30386242

RESUMO

Curcumin (CUR), a promising naturally occurring dietary compound, is commonly recognized as the potential anti-inflammatory agent. While the application of CUR was hampered by its low stability and poor systemic bioavailability, it has been suggested that the biological activities of CUR are intimately related to its metabolites. In the current investigation, we aimed to comparatively explore the anti-inflammatory effects of tetrahydrocurcumin (THC), octahydrocurcumin (OHC), and CUR, and to elucidate the underlying action mechanisms on experimental mice models of acute inflammation, i.e., xylene-induced ear edema, acetic acid-induced vascular permeability, and carrageenan-induced paw edema. The results showed that THC and OHC exerted significant and dose-dependent inhibitions on the formation of ear edema induced by xylene and paw edema provoked by carrageenan and inhibited the Evans blue dye leakage in peritoneal cavity elicited by acetic acid. Moreover, THC and OHC treatments were more effective than CUR in selectively inhibiting the expression of cyclooxygenase 2 (COX-2) and suppressing nuclear factor-κB (NF-κB) pathways via transforming growth factor ß activated kinase-1 (TAK1) inactivation in the carrageenan-induced mouse paw edema model.

5.
Zhongguo Zhong Yao Za Zhi ; 42(5): 989-997, 2017 Mar.
Artigo em Chinês | MEDLINE | ID: mdl-28994545

RESUMO

The study aims to analyze the varieties and standards of compositae medicinal plants used in Dai medicine. The results showed that there were 78 species (including varieties) compositae plants recorded in literatures, which belongs to 63 medicinal materials varieties. And 47 original plants (60.25%) were recorded in Chinese medicinal material standards. In those standards and literatures of Dai medicine, there are great differences in translated Chinese names, original plants, medicinal parts, and efficacy of medicinal plants. Therefore, the variety systematization and the quality standards of Dai medicine should be strengthened.


Assuntos
Asteraceae/classificação , Medicamentos de Ervas Chinesas/normas , Plantas Medicinais/classificação , Medicina Tradicional Chinesa , Fitoterapia , Padrões de Referência
6.
Nan Fang Yi Ke Da Xue Xue Bao ; 37(6): 721-729, 2017 Jun 20.
Artigo em Chinês | MEDLINE | ID: mdl-28669943

RESUMO

OBJECTIVE: To evaluate the development of the cerebral sulci and gyrus and cerebral blood flow in fetuses with tetralogy of Fallot (ToF) in the second and third trimesters using ultrasound imaging. METHODS: Forty fetuses (23-33+6 weeks) with ToF diagnosed using ultrasound imaging between December, 2015 and September, 2016 were analyzed in this study. The development of the cerebral sulci and gyrus was evaluated by measuring the parietal-occipital fissure (POF) depth, POF angle, sylvian fissure (SF) depth, SF width, uncovered insular width, calcarine fissure (CF) depth, hemisphere depth on the views of POF, SF and CF, uncovered insular ratio, biparietal diameter (BPD), and head circumference (HC). Cerebral hemodynamics were assessed by measuring the umbilical artery resistance index, umbilical artery pulsation index, middle cerebral artery pulsation index (MCA-PI), middle cerebral artery resistance index, the cerebral-to-placental resistance ratio, and the cerebroplacental ratio. RESULTS: In ToF fetuses, the POF depth, SF depth, CF depth, BPD, HC and hemisphere depth on the views of parietal-occipital sulcus and calcarine sulcus were significantly smaller than those in the control group (P<0.05). The middle cerebral artery resistance index, middle cerebral artery pulsation index, the cerebral-to-placental resistance ratio and the cerebroplacental ratio were also significantly lower in ToF fetuses than in the control group (P<0.05). CONCLUSION: The cerebral sulci and gyrus in ToF fetuses in second and third trimesters show underdevelopment compared with those in normal fetuses, and the changes in hemodynamics caused by abnormal cardiac structure might be one of the reasons for cerebral sulci and gyrus underdevelopment in fetuses with ToF.


Assuntos
Córtex Cerebral/efeitos dos fármacos , Circulação Cerebrovascular , Tetralogia de Fallot/diagnóstico por imagem , Ultrassonografia Pré-Natal , Feminino , Feto , Humanos , Artéria Cerebral Média , Gravidez
7.
Int Immunopharmacol ; 50: 270-278, 2017 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-28711783

RESUMO

ß-Patchoulene (ß-PAE), a tricyclic sesquiterpene isolated from the essential oil of the leaves and stems of Pogostemon cablin (Blanco) Benth., has been reported to have potent anti-inflammatory activity. The aim of this study was to evaluate the potential protective effect of ß-PAE on lipopolysaccharide (LPS)-induced acute lung injury (ALI) in mice and to illuminate the underlying mechanisms. ALI was induced by intracheal instillation of LPS into lung, and dexamethasone (DEX) was used as a positive control. Results indicated that pretreatment with ß-PAE significantly decreased the mortality rate of mice and lung W/D weight ratio, ameliorated lung pathological changes as compared to model group. Meanwhile, ß-PAE pretreatment markedly inhibited the increase of TNF-α, IL-6 and IL-1ß secretions in the bronchoalveolar lavage fluid, and prevented LPS-induced elevations of MPO activity and MDA level in the lung. Additionally, ß-PAE pretreatment significantly elevated miR-146a expression and suppressed the LPS-induced activation of NF-κB and expression of its mediated genes (TNF-α, IL-6 and IL-1ß). ß-PAE was also observed to markedly upregulate the Nrf2 and HO-1 expression and activate the antioxidant genes (NQO-1, GCLC and HO-1). Taken together, ß-PAE possessed protective effect against LPS-induced ALI, which might be associated with its differential regulation of NF-κB and Nrf2 activities and up-regulation of expression of miR-146a. The results rendered ß-PAE a promising anti-inflammatory agent worthy of further development into a pharmaceutical drug for the treatment of ALI.


Assuntos
Lesão Pulmonar Aguda/tratamento farmacológico , Anti-Inflamatórios/uso terapêutico , Pulmão/metabolismo , Fator 2 Relacionado a NF-E2/metabolismo , Sesquiterpenos/uso terapêutico , Animais , Citocinas/metabolismo , Dexametasona/imunologia , Modelos Animais de Doenças , Heme Oxigenase-1/genética , Heme Oxigenase-1/metabolismo , Humanos , Mediadores da Inflamação/metabolismo , Lipopolissacarídeos/imunologia , Pulmão/efeitos dos fármacos , Pulmão/patologia , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Camundongos , Camundongos Endogâmicos , MicroRNAs/genética , Fator 2 Relacionado a NF-E2/genética , NF-kappa B/metabolismo , Peroxidase/metabolismo , Pogostemon/imunologia , Sesquiterpenos de Guaiano , Transdução de Sinais
8.
Mol Med Rep ; 15(6): 4239-4246, 2017 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-28440419

RESUMO

Osthole is the primary active component of a number of herbal plants such as the Cnidium monnieri fruit. In traditional Chinese herb medicine, osthole is commonly used in combination with borneol to obtain improved pharmacological effects. The aim of the present study was to investigate the effect of borneol enantiomers on the pharmacokinetics of osthole. An appropriate high­performance liquid chromatography (HPLC) method was applied to determine the concentrations of osthole in plasma. Following oral administration of osthole alone or combined with borneol in rats, blood samples were collected and analyzed by HPLC. The results demonstrated that there were statistically significant differences in the pharmacokinetic parameters of osthole between osthole administration alone and co­administration with borneol. When combined with synthetic borneol, the AUC0­t, AUC0­∞ and Cmax of osthole increased by 48.153, 104.708 and 92.630%, respectively, while the CL/F decreased by 51.251%. When combined with (+)­borneol, the AUC0­t, AUC0­∞ and Cmax of osthole were increased by 61.561, 78.167, and 51.769%, respectively, while the CL/F decreased by 44.174% (P<0.01). In addition, when combined with (­)­borneol, the AUC0­t, AUC0­∞ and Cmax of osthole increased by 115.856, 167.786 and 271.289%, respectively, while the CL/F decreased by 60.686% (P<0.01). These results indicated that borneol may enhance gastrointestinal absorption and inhibit the metabolism of osthole. In addition, the promotional effect of (­)­borneol on the pharmacokinetic parameters of osthole was greater than that of (+)­borneol.


Assuntos
/farmacologia , Cumarínicos/farmacocinética , Administração Oral , Animais , Área Sob a Curva , Medicamentos de Ervas Chinesas/farmacocinética , Masculino , Medicina Tradicional Chinesa/métodos , Ratos , Ratos Sprague-Dawley
9.
Int J Mol Sci ; 18(3)2017 Feb 24.
Artigo em Inglês | MEDLINE | ID: mdl-28245556

RESUMO

Bleomycin (BLM), a family of anti-tumor drugs, was reported to exhibit severe side effects limiting its usage in clinical treatment. Therefore, finding adjuvants that enhance the anti-tumor effect and reduce the detrimental effect of BLM is a prerequisite. Chrysanthemum indicum, an edible flower, possesses abundant bioactivities; the supercritical-carbon dioxide fluid extract from flowers and buds of C. indicum (CISCFE) have strong anti-inflammatory, anti-oxidant, and lung protective effects. However, the role of CISCFE combined with BLM treatment on tumor-bearing mice remains unclear. The present study aimed to investigate the potential synergistic effect and the underlying mechanism of CISCFE combined with BLM in the treatment of hepatoma 22 (H22) tumor-bearing mice. The results suggested that the oral administration of CISCFE combined with BLM could markedly prolong the life span, attenuate the BLM-induced pulmonary fibrosis, suppress the production of pro-inflammatory cytokines (interleukin-6), tumor necrosis factor-α, activities of myeloperoxidase, and malondiadehyde. Moreover, CISCFE combined with BLM promoted the ascites cell apoptosis, the activities of caspases 3 and 8, and up-regulated the protein expression of p53 and down-regulated the transforming growth factor-ß1 by activating the gene expression of miR-29b. Taken together, these results indicated that CISCFE could enhance the anti-cancer activity of BLM and reduce the BLM-induced pulmonary injury in H22 tumor-bearing mice, rendering it as a potential adjuvant drug with chemotherapy after further investigation in the future.


Assuntos
Antibióticos Antineoplásicos/farmacologia , Bleomicina/farmacologia , Chrysanthemum/química , Extratos Vegetais/farmacologia , Animais , Antibióticos Antineoplásicos/efeitos adversos , Apoptose/efeitos dos fármacos , Bleomicina/efeitos adversos , Dióxido de Carbono , Caspase 3/metabolismo , Caspase 8/metabolismo , Linhagem Celular Tumoral , Citocinas/biossíntese , Modelos Animais de Doenças , Sinergismo Farmacológico , Fibrose , Extração Líquido-Líquido/métodos , Pulmão/metabolismo , Pulmão/patologia , Camundongos , MicroRNAs/genética , MicroRNAs/metabolismo , Neoplasias/tratamento farmacológico , Neoplasias/mortalidade , Neoplasias/patologia , Estresse Oxidativo/efeitos dos fármacos , Extratos Vegetais/química , Fator de Crescimento Transformador beta1/genética , Fator de Crescimento Transformador beta1/metabolismo , Carga Tumoral/efeitos dos fármacos , Proteína Supressora de Tumor p53/genética , Proteína Supressora de Tumor p53/metabolismo , Ensaios Antitumorais Modelo de Xenoenxerto
10.
J Med Food ; 20(2): 180-188, 2017 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-28146409

RESUMO

Constipation is a common disorder that is a significant source of morbidity among people around the world ranging from 2% to 28%. Dendrobium officinale Kimura et Migo is a traditional herbal medicine and health food used for tonicity of the stomach and promotion of body fluid production in China. This study aimed to prepare the ultrafine powder of Dendrobium officinale (UDO) and investigate its laxative effect and potential mechanism in mice with diphenoxylate-induced constipation. Results indicated that the mean diameter (d50) of UDO obtained by ball milling was 6.56 µm. UDO (62.5, 125, and 250 mg/kg, p.o.) could significantly enhance the gastrointestinal transit ratio and promote fecal output. Moreover, UDO treatment resulted in significant increases in the serum levels of acetylcholinesterase (AChE), gastrin (Gas), motilin (MTL), and substance P (SP), and obviously decreased serum contents of somatostatin (SS). Taken together, UDO, which can be easily obtained through milling to a satisfactory particle size, exhibited obvious laxative effect in diphenoxylate-induced constipated mice, and the mechanism might be associated with elevated levels of AChE, Gas, MTL, SP, and reduced production of SS. UDO has the potential for further development into an alternative effective diet therapy for constipation.


Assuntos
Constipação Intestinal/tratamento farmacológico , Dendrobium/química , Laxantes/administração & dosagem , Extratos Vegetais/administração & dosagem , Animais , Constipação Intestinal/metabolismo , Constipação Intestinal/fisiopatologia , Feminino , Gastrinas/metabolismo , Trânsito Gastrointestinal/efeitos dos fármacos , Humanos , Laxantes/química , Masculino , Camundongos , Camundongos Endogâmicos ICR , Motilina/metabolismo , Extratos Vegetais/química , Substância P/metabolismo
11.
Food Funct ; 7(11): 4545-4555, 2016 Nov 09.
Artigo em Inglês | MEDLINE | ID: mdl-27714005

RESUMO

Accumulating evidence has shown that chronic injection of d-galactose (d-gal) can mimic natural aging, with accompanying liver and brain injury. Oxidative stress and apoptosis play a vital role in the aging process. In this study, the antioxidant ability of polydatin (PD) was investigated using four established in vitro systems. An in vivo study was also conducted to investigate the possible protective effect of PD on d-gal-induced liver and brain damage. The results showed that PD had remarkable in vitro free radical scavenging activity on 2,2-diphenyl-1-picryl-hydrazyl (DPPH˙), 2,2'-azino-bis(3-ethylbenzo-thiazoline-6-sulfonic acid) (ABTS+˙) radical ions, and hydroxyl and superoxide anions. Results in vivo indicated that, in a group treated with d-gal plus PD, PD remarkably decreased the depression of body weight and organ indexes, reduced the levels of the serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST), and alleviated alterations in liver and brain histopathology. PD also significantly decreased the level of MDA and elevated SOD, GSH-Px, CAT activity and T-AOC levels in the liver and brain. In addition, the levels of inflammatory mediators, such as TNF-α, IL-1ß and IL-6 in serum were markedly reduced after PD treatment. Western blotting results revealed that PD treatment noticeably attenuated the d-gal-induced elevation of Bcl-2/Bax ratio and caspase-3 protein expression in liver and brain. Overall, our findings indicate that PD treatment could effectively attenuate d-gal-induced liver and brain damage, and the mechanism might be associated with decreasing the oxidative stress, inflammation and apoptosis caused by d-gal. PD holds good potential for further development into a promising pharmaceutical candidate for the treatment of age-associated diseases.


Assuntos
Anti-Inflamatórios/farmacologia , Antioxidantes/farmacologia , Encefalopatias/induzido quimicamente , Doença Hepática Induzida por Substâncias e Drogas/prevenção & controle , Galactose/toxicidade , Glucosídeos/farmacologia , Estilbenos/farmacologia , Envelhecimento , Animais , Anti-Inflamatórios/química , Antioxidantes/química , Apoptose/efeitos dos fármacos , Encefalopatias/prevenção & controle , Citocinas , Relação Dose-Resposta a Droga , Glucosídeos/administração & dosagem , Glucosídeos/química , Fígado , Malondialdeído , Camundongos , Estrutura Molecular , Estresse Oxidativo , Estilbenos/administração & dosagem , Estilbenos/química
12.
Early Hum Dev ; 100: 27-32, 2016 09.
Artigo em Inglês | MEDLINE | ID: mdl-27391870

RESUMO

BACKGROUND: Studies on motor performance and its early markers are rare in China, especially in very low birth weight (VLBW) infants. OBJECTIVE: Apart from the assessment of the inter-scorer agreement, we aimed to analyze to what extent the motor repertoire at 10 to 18weeks postterm was related to neonatal complications, and gross and fine motor performance at 12months after term. STUDY DESIGN: Exploratory prospective study. SUBJECTS: Seventy-four VLBW infants (58 males; mean gestational age=29weeks; mean birth weight=1252g). METHOD: Five-minute video recordings were performed at 10 to 18weeks after term; fidgety movements and the concurrent motor patterns (resulting in a motor optimality score) were assessed according to the Prechtl general movements assessment (GMA). The gross and fine motor performance was assessed by means of the Peabody Developmental Motor Scales, second edition, at 12months. RESULTS: Reliability was excellent. Pneumonia was associated with absent fidgety movements; the motor optimality score was lower in infants with pneumonia and/or bronchopulmonary dysplasia. Both absent fidgety movements and a lower motor optimality score were associated with a poor or very poor gross and fine motor performance at the 12-month-assessment. CONCLUSION: Both the assessment of fidgety movements and the evaluation of the concurrent motor repertoire contribute significantly to an identification of VLBW children with a poor gross and fine motor outcome at 12months. The results of this study document the need for an early identification of infants at high risk for a poor motor performance.


Assuntos
Recém-Nascido Prematuro/fisiologia , Recém-Nascido de muito Baixo Peso/fisiologia , Destreza Motora/fisiologia , Peso ao Nascer , Displasia Broncopulmonar/fisiopatologia , Desenvolvimento Infantil/fisiologia , China , Feminino , Idade Gestacional , Humanos , Lactente , Recém-Nascido , Doenças do Prematuro/fisiopatologia , Masculino , Movimento/fisiologia , Pneumonia/fisiopatologia , Estudos Prospectivos , Reprodutibilidade dos Testes , Fatores de Risco , Gravação em Vídeo
13.
Virol J ; 7: 232, 2010 Sep 16.
Artigo em Inglês | MEDLINE | ID: mdl-20843372

RESUMO

BACKGROUND: Some UL45 gene function of Herpesvirus was reported. While there was no any report of the duck enteritis virus (DEV) UL45 protein as yet. RESULTS: The UL45 gene and des-transmembrane domain of UL45 (named UL45Δ gene, 295-675bp of UL45) of DEV were amplified by PCR and subcloned into the prokaryotic expression vector pET-32a(+). The constructed recombinant plasmids were transformed into the host strain BL21(DE3) PLysS and induced by IPTG. SDS-PAGE analysis showed the UL45 gene couldn't express while UL45Δ gene was highly expressed. His Purify Kit or salting-out could purify the protein effectively. Using the purified protein to immunize New-Zealand rabbits and produce polyclonal antibody. The agar diffusion reaction showed the titer of antibody was 1:32. Western blot analysis indicated the purified rabbit anti-UL45Δ IgG had a high level of specificity and the UL45 gene was a part of DEV genome. The transcription phase study of UL45 gene showed that expression of UL45 mRNA was at a low level from 0 to 18 h post-infection (pi), then accumulated quickly at 24 h pi and peaked at 42 h pi. It can be detected till 72 h pi. Besides, western blot analysis of purified virion and different viral ingredients showed that the UL45 protein resided in the purified virion and the viral envelope. CONCLUSIONS: The rabbit anti-UL45Δ IgG was produced successfully and it can serve as a good tool for penetrating studies of the function of DEV UL45 protein. The transcription phase and protein characteristics analysis indicated that DEV UL45 gene was a late gene and UL45 protein may be a viral envelope protein.


Assuntos
Patos/virologia , Perfilação da Expressão Gênica , Regulação Viral da Expressão Gênica , Herpesviridae/genética , Proteínas Estruturais Virais/biossíntese , Animais , Anticorpos Antivirais/imunologia , Anticorpos Antivirais/isolamento & purificação , Clonagem Molecular , Expressão Gênica , Coelhos , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Proteínas Recombinantes/isolamento & purificação , Fatores de Tempo , Transcrição Genética , Proteínas Estruturais Virais/imunologia , Vírion/química
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