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1.
Univ. sci ; 23(2): 171-189, May-Aug. 2018. tab, graf
Article in English | LILACS, COLNAL | ID: biblio-979544

ABSTRACT

Abstract Due to the health risks for both humans and living beings caused by polycyclic aromatic hydrocarbons (PAHs), the monitoring of these compounds in environmental matrices is mandatory. This work proposes an analytical method for analyzing anthracene (AN) and benzo[a]pyrene (BaP), two of the most representative PAHs, at ultra-trace concentrations in water, employing direct injection of large volumes of samples coupled with reversed-phase high-performance liquid chromatography. For this purpose, principal component analysis was used to examine the behavior of AN and BaP within the chromatographic system. Results showed that AN and BaP chromatographic behavior can be described by three models representing their identification, the quantification of AN and that of BaP, respectively. The factors affecting the obtained models, such as the injection volume, column temperature, flow rate, strength of the mobile phase, and the excitation and emission wavelengths, were examined and optimized by means of design of experiments. Finally, the analytical method was validated, obtaining promising limits of detection and quantification. The developed analytical method was demonstrated to be useful for a sensitive analysis of the target analytes in relatively clean natural water matrices.


Resumen Los hidrocarburos aromáticos policíclicos (HAPs) causan problemas en la salud de los seres humanos y seres vivos, por lo que se requiere un monitoreo de estos compuestos en matrices ambientales. Este trabajo propone un método analítico para analizar el antraceno (AN) y el benzopireno (BAP), los hidrocarburos más representativos en concentraciones de ultra trazas en el agua, empleando inyección directa en grandes volúmenes en muestras acopladas a la fase inversa con cromatografía líquida de alto rendimiento. Por tal razón, se utilizó el análisis de componentes principales para examinar el comportamiento de AN y BAP en el sistema cromatográfico. Los resultados mostraron que el comportamiento cromatográfico de AN y BAP podría describirse por medio de tres modelos que representan su identificación, la cuantificación de AN y de BAP, respectivamente. Se examinaron los factores que afectan a los modelos obtenidos, como el volumen de inyección, la temperatura de la columna, la tasa de flujo, la fuerza de la fase móvil, y las longitudes de las ondas de excitación y emisión, y se optimizaron mediante el diseño de experimentos. Finalmente, se validó el método analítico, obteniendo límites de detección y cuantificación. Se demostró que el método analítico desarrollado fue útil para el análisis sensible de los analitos en matrices de agua natural relativamente limpia.


Resumo Devido aos riscos para a saúde tanto para humanos como para os seres vivos em geral causados pelos hidrocarbonos aromáticos policíclicos (HAPs), o monitoramento de estes compostos em matrizes ambientais é prioritário. Este trabalho propõem um método analítico para analisar antraceno (AN) e benzo[a]pireno (BaP), dois dos hidrocarbonos mais representativos, em concentrações de ultra traços em água, empregando injeções diretas de grandes volumes de amostra acoplada a cromatografia líquida de alta eficiência em fase reversa. Usando Análises por Componentes Principais e desenho experimental, foram avaliados os efeitos de diversos fatores que afetam o sistema cromatográfico, tais como o volume de injeção, a temperatura da coluna, fluxo, forca da fase móvel e comprimentos de onda de excitação e emissão. Os resultados demonstraram que o comportamento cromatográfico de AN e BaP pode ser descrito por meio de três que representam sua identificação, quantificação de AN e de BaP, respectivamente. Os resultados mostraram que o comportamento cromatográfico de NA e BAP poderia ser descrito por meio de três modelos que representam sua identificação, a quantificação de NA e de BAP, respectivamente. Examinaram-se os fatores que afetam aos modelos obtidos, como o volume de injeção, a temperatura da coluna, a taxa de fluxo, a forca da fase móvel, e as longitudes das ondas de excitação e emissão, e se otimizaram mediante o desenho experimental. Finalmente, se validou o m todo analítico, obtendo os limites de detecção e quantificação. O método analítico desenvolvido demonstrou ser útil para uma análise sensível para os compostos de interesse em matrizes de água natural relativamente limpas.


Subject(s)
Polycyclic Aromatic Hydrocarbons , Environmental Pollution , Anthracenes
2.
Biosci. j. (Online) ; 34(4): 888-898, july/aug. 2018. tab, graf
Article in English | LILACS | ID: biblio-967032

ABSTRACT

The efficiency of co-application of Eisenia fetida and ryegrass was evaluated in a process called earthworm-assisted phytoremediation. Anthracene was used as a model compound for polycyclic aromatic hydrocarbons (PAHs). The experiments were conducted on a loamy soil in greenhouse conditions. At the end of the experiment, the soil samples were analyzed for residual anthracene by HPLC. Results showed that, phytoremediation using ryegrass could remove 81% of anthracene; however, the rate of removal was 92% when E. fetida was applied simultaneously. E. fetida alone could also remove the initial concentration of anthracene by 40%. Although ryegrass itself could remove anthracene significantly, the employment of earthworm, together with plant was more efficient than each of them individually. The application of E. fetida could also enhance the growth parameters of ryegrass significantly. In comparison to the control, the presence of E. fetida increased plant dry weight (7.8%), root length (47%), shoots length (32%), and root volume (12%). The number of live earthworms was also increased in the planted pots, indicating the helpfulness of the plant for survival of the earthworm in the PAH-contaminated soil. Although plant and earthworm use completely different mechanisms for anthracene degradation, they improve efficiency and survival of the three-component-system.


A eficiência da co-aplicação de Eisenia fetida e azevém foi avaliada em um processo denominado fitorremediação assistida por minhocas. O antraceno foi usado como um composto modelo para hidrocarbonetos aromáticos policíclicos (PAHs). Os experimentos foram conduzidos em um solo argiloso em condições de estufa. No final da experiência, as amostras de solo foram analisadas quanto ao antraceno residual por HPLC. Os resultados mostraram que, a fitorremediação com azevém pode remover 81% do antraceno; no entanto, a taxa de remoção foi de 92% quando E. fetida foi aplicada simultaneamente. E. fetida sozinha também foi capaz de remover a concentração inicial de antraceno em 40%. Embora o próprio azevém pudesse remover significativamente o antraceno, o emprego da minhoca, juntamente com a planta, foi mais eficiente do que cada um deles individualmente. A aplicação de E. fetida também pode melhorar significativamente os parâmetros de crescimento do azevém. Em comparação com o controle, a presença de E. fetida aumentou o peso seco da planta (7,8%), o comprimento da raiz (47%), o comprimento da parte aérea (32%) e o volume radicular (12%). O número de minhocas vivas também aumentou nos vasos plantados, indicando a utilidade da planta para a sobrevivência da minhoca no solo contaminado com PAH. Embora plantas e minhocas usem mecanismos completamente diferentes para a degradação do antraceno, eles melhoram a eficiência e a sobrevivência do sistema de três componentes.


Subject(s)
Oligochaeta , Biodegradation, Environmental , Anthracenes , Lolium , Hydrocarbons
3.
Chinese Medical Journal ; (24): 1719-1724, 2016.
Article in English | WPRIM | ID: wpr-251316

ABSTRACT

<p><b>BACKGROUND</b>An acute respiratory distress syndrome (ARDS) is still one of the major challenges in critically ill patients. This study aimed to investigate the effect of inhibiting c-Jun N-terminal kinase (JNK) on ARDS in a lipopolysaccharide (LPS)-induced ARDS rat model.</p><p><b>METHODS</b>Thirty-six rats were randomized into three groups: control, LPS, and LPS + JNK inhibitor. Rats were sacrificed 8 h after LPS treatment. The lung edema was observed by measuring the wet-to-dry weight (W/D) ratio of the lung. The severity of pulmonary inflammation was observed by measuring myeloperoxidase (MPO) activity of lung tissue. Moreover, the neutrophils in bronchoalveolar lavage fluid (BALF) were counted to observe the airway inflammation. In addition, lung collagen accumulation was quantified by Sircol Collagen Assay. At the same time, the pulmonary histologic examination was performed, and lung injury score was achieved in all three groups.</p><p><b>RESULTS</b>MPO activity in lung tissue was found increased in rats treated with LPS comparing with that in control (1.26 ± 0.15 U in LPS vs. 0.77 ± 0.27 U in control, P < 0.05). Inhibiting JNK attenuated LPS-induced MPO activity upregulation (0.52 ± 0.12 U in LPS + JNK inhibitor vs. 1.26 ± 0.15 U in LPS, P < 0.05). Neutrophils in BALF were also found to be increased with LPS treatment, and inhibiting JNK attenuated LPS-induced neutrophils increase in BALF (255.0 ± 164.4 in LPS vs. 53 (44.5-103) in control vs. 127.0 ± 44.3 in LPS + JNK inhibitor, P < 0.05). At the same time, the lung injury score showed a reduction in LPS + JNK inhibitor group comparing with that in LPS group (13.42 ± 4.82 vs. 7.00 ± 1.83, P = 0.001). However, the lung W/D ratio and the collagen in BALF did not show any differences between LPS and LPS + JNK inhibitor group.</p><p><b>CONCLUSIONS</b>Inhibiting JNK alleviated LPS-induced acute lung inflammation and had no effects on pulmonary edema and fibrosis. JNK inhibitor might be a potential therapeutic medication in ARDS, in the context of reducing lung inflammatory.</p>


Subject(s)
Animals , Anthracenes , Therapeutic Uses , Collagen , Metabolism , JNK Mitogen-Activated Protein Kinases , Metabolism , Lipopolysaccharides , Toxicity , Lung , Metabolism , Pathology , Male , Rats , Respiratory Distress Syndrome , Drug Therapy , Signal Transduction
4.
Article in English | WPRIM | ID: wpr-285258

ABSTRACT

Swelling-activated chloride currents (ICl.swell) are thought to play a role in several physiologic and pathophysiologic processes and thus represent a target for therapeutic approaches. However, the mechanism of ICl.swell regulation remains unclear. In this study, we used the whole-cell patch-clamp technique to examine the role of protein kinase C (PKC) in the regulation of ICl.swell in human atrial myocytes. Atrial myocytes were isolated from the right atrial appendages of patients undergoing coronary artery bypass and enzymatically dissociated. ICl.swell was evoked in hypotonic solution and recorded using the whole-cell patch-clamp technique. The PKC agonist phorbol dibutyrate (PDBu) enhanced ICl.swell in a concentration-dependent manner, which was reversed in isotonic solution and by a chloride current inhibitor, 9-anthracenecarboxylicacid. Furthermore, the PKC inhibitor bis-indolylmaleimide attenuated the effect and 4α-PDBu, an inactive PDBu analog, had no effect on ICl.swell. These results, obtained using the whole-cell patch-clamp technique, demonstrate the ability of PKC to activate ICl,swell in human atrial myocytes. This observation was consistent with a previous study using a single-channel patch-clamp technique, but differed from some findings in other species.


Subject(s)
Anthracenes , Pharmacology , Chloride Channels , Metabolism , Chlorides , Metabolism , Culture Media , Metabolism , Pharmacology , Dose-Response Relationship, Drug , Evoked Potentials , Physiology , Heart Atria , Cell Biology , Metabolism , Humans , Hypotonic Solutions , Metabolism , Pharmacology , Indoles , Pharmacology , Ion Transport , Maleimides , Pharmacology , Myocytes, Cardiac , Cell Biology , Metabolism , Patch-Clamp Techniques , Phorbol 12,13-Dibutyrate , Pharmacology , Primary Cell Culture , Protein Kinase C , Metabolism
5.
Chinese Medical Journal ; (24): 2853-2860, 2016.
Article in English | WPRIM | ID: wpr-230868

ABSTRACT

<p><b>BACKGROUND</b>Cathepsin L (CatL) is a cysteine protease with strong matrix degradation activity that contributes to photoaging. Mannose phosphate-independent sorting pathways mediate ultraviolet A (UVA)-induced alternate trafficking of CatL. Little is known about signaling pathways involved in the regulation of UVA-induced CatL expression and activity. This study aims to investigate whether a single UVA irradiation affects CatL expression and activity and whether mitogen-activated protein kinase (MAPK)/activator protein-1 (AP-1) pathway is involved in the regulation of UVA-induced CatL expression and activity in human dermal fibroblasts (HDFs).</p><p><b>METHODS</b>Primary HDFs were exposed to UVA. Cell proliferation was determined by a cell counting kit. UVA-induced CatL production and activity were studied with quantitative real-time reverse transcription polymerase chain reaction (RT-PCR), Western blotting, and fluorimetric assay in cell lysates collected on three consecutive days after irradiation. Time courses of UVA-activated JNK and p38MAPK signaling were examined by Western blotting. Effects of MAPK inhibitors and knockdown of Jun and Fos on UVA-induced CatL expression and activity were investigated by RT-PCR, Western blotting, and fluorimetric assay. Data were analyzed by one-way analysis of variance.</p><p><b>RESULTS</b>UVA significantly increased CatL gene expression, protein abundance, and enzymatic activity for three consecutive days after irradiation (F = 83.11, 56.14, and 71.19, respectively; all P < 0.05). Further investigation demonstrated phosphorylation of JNK and p38MAPK activated by UVA. Importantly, inactivation of JNK pathway significantly decreased UVA-induced CatL expression and activity, which were not affected by p38MAPK inhibition. Moreover, knockdown of Jun and Fos significantly attenuated basal and UVA-induced CatL expression and activity.</p><p><b>CONCLUSIONS</b>UVA enhances CatL production and activity in HDFs, probably by activating JNK and downstreaming AP-1. These findings provide a new possible molecular approach for antiphotoaging therapy.</p>


Subject(s)
Anthracenes , Pharmacology , Cathepsin L , Metabolism , Cells, Cultured , Child , Child, Preschool , Enzyme Inhibitors , Pharmacology , Extracellular Signal-Regulated MAP Kinases , Fibroblasts , Cell Biology , Metabolism , Radiation Effects , Humans , Imidazoles , Pharmacology , MAP Kinase Signaling System , Radiation Effects , Oncogene Proteins v-fos , Genetics , Metabolism , Proto-Oncogene Proteins c-jun , Genetics , Metabolism , Pyridines , Pharmacology , Skin , Cell Biology , Ultraviolet Rays
6.
Article in Chinese | WPRIM | ID: wpr-299856

ABSTRACT

Study a method for the detemination of the content of polysaccharides in Gentiana farreri, and analysis of the content of polysaccharides from different producing areas. The results showed that using the anthrone-sulfuric acid method, simple operation, accurate result. Sample was measured at 620 nm absorbance after anthrone-sulfuric acid color, at this wavelength, solution absorption and glucose showed a good linear relationship; The linearity was in the range of 0.01-0.07 g x L(-1) (r = 0.996 7). The recovery rate was 99.41%, with RSD of 2.0%. Considering the experimental conditions, to determine the solid-liquid ratio 1:60, extracting time 50 min, concentration of ethanol 80%. The mass fraction of polysaccharides was the highest to reached 0.743% in G. farreri from Gansu Xiahe. This experiment has laid a good foundation for further study on G. farreri.


Subject(s)
Anthracenes , Chemistry , Chemistry Techniques, Analytical , Methods , Gentiana , Chemistry , Geography , Linear Models , Polysaccharides , Reproducibility of Results , Sulfuric Acids , Chemistry , Time Factors
7.
Article in Chinese | WPRIM | ID: wpr-244542

ABSTRACT

<p><b>OBJECTIVE</b>To understand the comorbid characteristics and distribution of combined treatment of Chinese and Western medicine in depressive patients.</p><p><b>METHOD</b>The descriptive statistic method and association rule were used to analyze the data from 19 general hospitals with 3-A level in China.</p><p><b>RESULT</b>Among the depressive disorder, the most frequent co-morbid physical diseases included hypertension (24.67%), coronary heart disease (16.10%) and cerebral infarction (12.89%), and the proportion of comorbid changes with the increasing age, from 6.51% to 12.55%, 16.33% and 12.47% for hypertension; from 2.79% to 5.69%, 10.17% and 14.22% for coronary heart disease; from 3.72%, 6.27%, 7.70% and 12.25% for cerebral infarction. The use frequency of the antidepressants is 77.18%, and the use frequency of flupentixol & melitracen is 20.95%. The use frequency of Huoxue Huayu Tongluo of traditional Chinese medicine is 59.97%, with that of 27.91% for Ginkgo biloba extract The combined use frequency of Huoxue Huayu Tongluo of TCM and the antidepressants is the highest, especially for the combined use of Shuxuening injection and fluoxetine.</p><p><b>CONCLUSION</b>The most frequent comorbid diseases of depression include three kinds of diseases, such as hypertension, coronary heart disease and cerebral infarction, and its proportion gradually increased with the growth of age. The single use frequency of flupentixol & melitracen and G. biloba extract is the highest, while the combined use of Shuxuening injection and fluoxetine is the highest.</p>


Subject(s)
Adolescent , Adult , Aged , Aged, 80 and over , Anthracenes , Therapeutic Uses , Antidepressive Agents , Therapeutic Uses , Child , Child, Preschool , China , Combined Modality Therapy , Methods , Depressive Disorder , Drug Therapy , Drug Combinations , Drugs, Chinese Herbal , Therapeutic Uses , Female , Fluoxetine , Therapeutic Uses , Flupenthixol , Therapeutic Uses , Ginkgo biloba , Chemistry , Humans , Infant , Infant, Newborn , Male , Medicine, Chinese Traditional , Methods , Middle Aged , Plant Extracts , Therapeutic Uses , Young Adult
8.
Article in Chinese | WPRIM | ID: wpr-343677

ABSTRACT

<p><b>OBJECTIVE</b>To study the role of c-jun N-terminal kinase (JNK) signaling pathway in chondrocyte apoptosis induced by nitric oxide (NO) using NO donor sodium nitroprusside (SNP) and JNK inhibitor SP600125.</p><p><b>METHODS</b>Articular chondrocytes were separated from New Zealand rabbits aged 3 weeks by mechanical digestion and enzyme digestion and identified by toluidine blue staining, and then the chondrocytes were treated with SNP and SP600125 for 24 h. The cell apoptosis was evaluated by Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) flow cytometry and terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-biotin nick end labeling (TUNEL), and the expression levels of nuclear factor-kappa B (NF-κB) p65 and p53 were measured by western blot.</p><p><b>RESULTS</b>Compared with those in control group, the early apoptotic rate of SNP-treated chondrocytes increased as the concentration of SNProse, exhibiting a concentration dependency (P < 0.05), and the expression levels of NF-κB p65 and p53 also increased (P < 0.05); JNK inhibitor SP600125 inhibited these increases (P < 0.05).</p><p><b>CONCLUSION</b>JNK signaling pathway plays an important role in NO-induced chondrocyte apoptosis. JNK inhibitor SP600125 can reduce NO-induced apoptosis and expression of NF-κB p65 and p53 in articular chondrocytes of rabbits in a concentration-dependent manner.</p>


Subject(s)
Animals , Anthracenes , Pharmacology , Apoptosis , Cells, Cultured , Chondrocytes , Metabolism , Pathology , MAP Kinase Signaling System , NF-kappa B , Metabolism , Nitric Oxide , Pharmacology , Rabbits , Transcription Factor RelA , Metabolism , Tumor Suppressor Protein p53 , Metabolism
9.
Article in Chinese | WPRIM | ID: wpr-749216

ABSTRACT

OBJECTIVE@#To investigate the extracellular release of high mobility group box 1 (HMGB1) in laryngeal Hep-2 carcinoma cells induced by hypoxia and its possible mechanism.@*METHOD@#The changes of HMGB1 concentration in the culture medium as well as HMGB1 protein and mRNA expression in Hep-2 cells were investigated after the cells were cultured with 1% O2 for different durations. Inhibitory effects of MAPK pathway inhibitors (PD98059. SP600125, and SB202190) and nuclear NF-kappaB pathway inhibitor (PDTC) with various concentrations on extracellular HMGB1 release were observed in hypoxia-induced Hep-2 cells. The HMGB1 concentration and HMGB1 protein expression were measured by enzyme-linked immunosorbent assay (ELISA) and western blot, respectively. The HMGB1 mRNA expression was determined by real-time quantitative PCR(RT-PCR).@*RESULT@#The HMGB1 concentration in the culture medium and the HMGB1 protein expression in Hep-2 cells increased after the cells were subjected to hypoxia culture for 12 h in a time-dependent manner. The level of HMGB1 mRNA expression in Hep-2 cells increased after the cells were induced by hypoxia for 6h PD98059 and SP600125 with 20 micromol/ L and PDTC with 50 mg/L partly inhibited extracellular release of HMGB1 in hypoxia-cultured Hcp-2 cells.@*CONCLUSION@#Hypoxia induces laryngeal carcinoma cells to release HMGH1. which may be related to MAPK/NF-kappaB signaling pathway.


Subject(s)
Anthracenes , Cell Hypoxia , Cell Line, Tumor , Flavonoids , HMGB1 Protein , Metabolism , Humans , Imidazoles , MAP Kinase Signaling System , NF-kappa B , Metabolism , Protein Kinase Inhibitors , Pyridines , Pyrrolidines , RNA, Messenger , Genetics , Thiocarbamates
10.
Article in English | WPRIM | ID: wpr-319400

ABSTRACT

<p><b>OBJECTIVE</b>To study the effect of histone deacetylase inhibitors trichostatin A (TSA) and LBH589 on the growth of human renal cell carcinoma OS-RC-2 cells in vitro and explore the underlying molecular mechanism.</p><p><b>METHODS</b>OS-RC-2 cells were treated with LBH589 or TSA with or without SP600125 pretreatment, and the cell viability was measured by MTT assay. The changes of cell cycle distribution and apoptosis of OS-RC-2 cells were examined by flow cytometry, and the expressions of c-Jun, p-c-Jun, Bcl-2, and Bax were quantified by Western blotting.</p><p><b>RESULTS</b>TSA and LBH589 both inhibited the growth of OS-RC-2 cells in a dose- and time-dependent manner. TSA at 1 µnmol/L and LBH589 at 50 nmol/L caused obvious cell cycle arrest in G2/M phase and cell apoptosis, and significantly increased the protein levels of phosphorylated c-Jun. TSA treatment obviously increased Bax expression but decreased Bcl2 expression in the cells. The growth inhibitory effect of TSA was attenuated by the JNK inhibitor SP600125 in OS-RC-2 cells. TSA-induced phosphorylation of c-Jun and Bax upregulation was partially counteracted by SP600125.</p><p><b>CONCLUSION</b>TSA and LBH589 can cause cell cycle arrest and induce apoptosis in OS-RC-2 cells, in which process P-JNK pathway plays an important role.</p>


Subject(s)
Anthracenes , Pharmacology , Antineoplastic Agents , Pharmacology , Apoptosis , Carcinoma, Renal Cell , Metabolism , Pathology , Cell Cycle , Cell Line, Tumor , Cell Survival , Dose-Response Relationship, Drug , Histone Deacetylase Inhibitors , Pharmacology , Humans , Hydroxamic Acids , Pharmacology , Indoles , Pharmacology , JNK Mitogen-Activated Protein Kinases , Metabolism , Kidney Neoplasms , Metabolism , Pathology , MAP Kinase Signaling System , Phosphorylation , Proto-Oncogene Proteins c-bcl-2 , Metabolism , bcl-2-Associated X Protein , Metabolism
11.
Chinese Medical Journal ; (24): 2067-2073, 2013.
Article in English | WPRIM | ID: wpr-273036

ABSTRACT

<p><b>BACKGROUND</b>C-Jun N-terminal kinase (JNK) signaling pathway and ankylosis gene (ANK) play a critical role in endplate chondrocytes degeneration. The purpose of this study was to investigate whether the expression levels of ANK was associated with the activation of JNK.</p><p><b>METHODS</b>Cartilage endplates of 49 patients were divided into the control group (n = 19) and the experimental group (n = 30). The patients in the control group were graded 0 and those in the experimental group were graded I-III according to Miller's classification. Endplate chondrocytes were isolated by enzyme digestion and cultured in vitro. The inverted phase contrast microscope, teluidine blue staining, HE staining, real time RT-PCR, and MTT were used to observe morphological appearances, biological characteristics, and growth curve of endplate chondrocytes from the cartilage endplate of the two groups. Real time RT-PCR and Western blotting were used to analyze the mRNA and protein expression levels of associated factors in the degeneration process in the cultured endplate chondrocytes with or without subjected SP600125.</p><p><b>RESULTS</b>The expression levels of type II collagen, aggrecan, and ANK in endplate chondrocytes of experimental group were lower than that of control group and phosphorylation level of JNK in the experimental group which was higher than that in the control group. Application of JNK phosphorylation inhibitor to degeneration chondrocytes resulted in a marked decrease in the phosphorylation level of JNK and a significant increase in the expression levels of type II collagen, aggrecan, and ANK.</p><p><b>CONCLUSION</b>The degeneration of the human cervical endplate chondrocytes might be promoted by JNK phosphorylation by down-regulating the expression of ANK.</p>


Subject(s)
Adult , Aged , Anthracenes , Pharmacology , Cells, Cultured , Cervical Vertebrae , Metabolism , Pathology , Chondrocytes , Metabolism , Pathology , Down-Regulation , Female , Humans , JNK Mitogen-Activated Protein Kinases , Metabolism , Male , Middle Aged , Phosphate Transport Proteins , Genetics , Physiology , Phosphorylation
12.
Article in English | WPRIM | ID: wpr-727479

ABSTRACT

Vascular cell adhesion molecule 1 (VCAM-1), intercellular adhesion molecule 1 (ICAM-1), P- and E-selectin play a pivotal role for initiation of atherosclerosis. Ginsenoside, a class of steroid glycosides, is abundant in Panax ginseng root, which has been used for prevention of illness in Korea. In this study, we investigated the mechanism(s) by which ginsenoside Rg2 may inhibit VCAM-1 and ICAM-1 expressions stimulated with lipopolysaccharide (LPS) in human umbilical vein endothelial cell (HUVEC). LPS increased VCAM-1 and ICAM-1 expression. Ginsenoside Rg2 prevented LPS-mediated increase of VCAM-1 and ICAM-1 expression. On the other hand, JSH, a nuclear factor kappa B (NF-kappaB) inhibitor, reduced both VCAM-1 and ICAM-1 expression stimulated with LPS. SB202190, inhibitor of p38 mitogen-activated protein kinase (p38 MAPK), and wortmannin, phosphatidylinositol 3-kinase (PI3-kinase) inhibitor, reduced LPS-mediated VCAM-1 but not ICAM-1 expression. PD98059, inhibitor of mitogen-activated protein kinase kinase/extracellular signal-regulated kinase (MEK/ERK) did not affect VCAM-1 and ICAM-1 expression stimulated with LPS. SP600125, inhibitor of c-Jun N-terminal kinase (JNK), reduced LPS-mediated ICAM-1 but not VCAM-1 expression. LPS reduced IkappaBalpha (IkappaBalpha) expression, in a time-dependent manner within 1 hr. Ginsenoside Rg2 prevented the decrease of IkappaBalpha expression stimulated with LPS. Moreover, ginsenoside Rg2 reduced LPS-mediated THP-1 monocyte adhesion to HUVEC, in a concentration-dependent manner. These data provide a novel mechanism where the ginsenoside Rg2 may provide direct vascular benefits with inhibition of leukocyte adhesion into vascular wall thereby providing protection against vascular inflammatory disease.


Subject(s)
Androstadienes , Anthracenes , Atherosclerosis , E-Selectin , Endothelial Cells , Flavonoids , Ginsenosides , Glycosides , Hand , Humans , I-kappa B Proteins , Imidazoles , Intercellular Adhesion Molecule-1 , JNK Mitogen-Activated Protein Kinases , Korea , Leukocytes , Monocytes , NF-kappa B , Panax , Phosphatidylinositol 3-Kinase , Phosphotransferases , Protein Kinases , Pyridines , Umbilical Veins , Vascular Cell Adhesion Molecule-1
13.
Article in English | WPRIM | ID: wpr-727478

ABSTRACT

Lysolipids such as LPA, S1P and SPC have diverse biological activities including cell proliferation, differentiation, and migration. We investigated signaling pathways of LPA-induced contraction in feline esophageal smooth muscle cells. We used freshly isolated smooth muscle cells and permeabilized cells from cat esophagus to measure the length of cells. Maximal contraction occurred at 10(-6) M and the response peaked at 30s. To identify LPA receptor subtypes in cells, western blot analysis was performed with antibodies to LPA receptor subtypes. LPA1 and LPA3 receptor were detected at 50 kDa and 44 kDa. LPA-induced contraction was almost completely blocked by LPA receptor (1/3) antagonist KI16425. Pertussis toxin (PTX) inhibited the contraction induced by LPA, suggesting that the contraction is mediated by a PTX-sensitive G protein. Phospholipase C (PLC) inhibitors U73122 and neomycin, and protein kinase C (PKC) inhibitor GF109203X also reduced the contraction. The PKC-mediated contraction may be isozyme-specific since only PKCepsilon antibody inhibited the contraction. MEK inhibitor PD98059 and JNK inhibitor SP600125 blocked the contraction. However, there is no synergistic effect of PKC and MAPK on the LPA-induced contraction. In addition, RhoA inhibitor C3 exoenzyme and ROCK inhibitor Y27632 significantly, but not completely, reduced the contraction. The present study demonstrated that LPA-induced contraction seems to be mediated by LPA receptors (1/3), coupled to PTX-sensitive G protein, resulting in activation of PLC, PKC-epsilon pathway, which subsequently mediates activation of ERK and JNK. The data also suggest that RhoA/ROCK are involved in the LPA-induced contraction.


Subject(s)
Amides , Animals , Anthracenes , Antibodies , Blotting, Western , Cats , Cell Proliferation , Contracts , Esophagus , Estrenes , Flavonoids , GTP-Binding Proteins , Indoles , Isoxazoles , Maleimides , Muscle, Smooth , Myocytes, Smooth Muscle , Neomycin , Pertussis Toxin , Propionates , Protein Kinase C , Pyridines , Pyrrolidinones , Receptors, Lysophosphatidic Acid , Type C Phospholipases
14.
Int. j. morphol ; 30(1): 284-289, mar. 2012. ilus
Article in English | LILACS | ID: lil-638801

ABSTRACT

Peroxisomicine A1 (PA1), one of the toxins isolated from seeds of plants of the Karwinskia genus, whose targets organs are the liver, kidney, and lungs. There is a selective toxicity in vitro to cancer-cell lines derived from the lungs, liver, and colon, compared to normal cell lines. PA1 caused apoptosis in several cancer-cell lines in culture. In toxic doses to rodents, it causes extensive apoptosis in the liver, kidney, and lungs. In our study we were interested in evaluating, for the first time, the morphological effects of administration of PA1 to implanted TC-1 cells and in the target organs in vivo. The TC-1 cells were cultured and injected into the hind limb of C57BL-6 mice. The animals were divided into 3 groups; those treated with four doses of 1 mg/kg each of PA1, the untreated control, and the vehicle-control groups. All mice were killed 10 days after cell implantation. Samples were obtained from TC-1 cells at the implantation site and from the liver, kidney, and lungs. The samples were processed for examination under light and electron microscopy. In the PA1-treated group, the TC-1 cells had necrosis, whereas in the control groups the tumor cells were undamaged. The target organs did not show any lesions. We demonstrated for the first time that there is a selective toxic effect of PA1 on the TC-1 cells in vivo.


Peroxisomicina A1 (PA1), una de las toxinas aisladas de las semillas de plantas del género Karwinskia, cuyos órganos blanco son hígado, riñón y pulmón. Hay una toxicidad selectiva in vitro contra líneas celulares cancerosas derivadas de pulmón, hígado y colon, comparadas con líneas celulares normales. PA1 causa apoptosis en varias líneas celulares malignas en cultivo. En dosis tóxicas a roedores, causa extensa apoptosis en hígado, riñón y pulmón. En nuestro estudio, estuvimos interesados en evaluar por primera vez los efectos morfológicos de la administración temprana de PA1 sobre células TC-1 implantadas y los órganos blanco in vivo. Las células TC-1 fueron cultivadas e implantadas en la extremidad posterior de ratones C57BL-6. Los animales fueron divididos en tres grupos: tratado con cuatro dosis de 1 mg/kg de peso de PA1, control sin tratamiento y control vehículo. Todos los animales fueron sacrificados 10 días posterior al implante de las células. Se colectaron muestras del sitio del implante de las células TC-1 y de hígado, riñón y pulmón. Las muestras fueron procesadas para su análisis a microscopía de luz y microscopia electrónica de transmisión. En el grupo tratado con PA1, las células TC-1 presentaron necrosis, mientras que en los grupos control las células tumorales se observaron sin daño. Los órganos blanco no mostraron lesión alguna. Demostramos por primera vez que existe un efecto tóxico selectivo de PA1 sobre las células TC-1 in vivo.


Subject(s)
Rats , Anthracenes/administration & dosage , Anthracenes/therapeutic use , Necrosis/chemically induced , Necrosis/veterinary , Cytostatic Agents/administration & dosage , Cytostatic Agents/therapeutic use , Mice , Molecular Targeted Therapy , Toxicity Tests/methods
15.
Article in Chinese | WPRIM | ID: wpr-329894

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the protective effects and mechanism of SP600125-specificity inhibitor of c-Jun N-terminal kinase (JNK)on lung ischemia /reperfusion injury in rats.</p><p><b>METHODS</b>The unilateral lung ischemia/reperfusion model was replicated in vivo. Rats were randomly divided into three groups (n = 10): control group, ischemia/reperfusion group ( I/R group) and ischemia/reperfusion + SP600125 group (SP600125 group). The lung tissues sampled at the end of each experiment were assayed for wet/dry weight ratio (W/D),the injured alveoli rate (IAR), the expression of phosphorylation JNK (p-JNK) and JNK protein were detected by Western blot, the expression of Bcl-2, Bax, Caspase3 protein were detected by immunocytochemistry techniques, the pneumocyte apoptosis index (AI) was detected by terminal deoxynucleotidyl transferase mediated dUTP nick end abeling(TUNEL), the ultrastructure changes were observed under electron microscope.</p><p><b>RESULTS</b>Compared to I/R group, the expression of p-JNK, Bcl-2, Bax and caspase-3 protein were markedly decreased (all P < 0.01), the expression of Bcl-2 protein and the ratio of Bcl-2/Bax were markedly increased in SP600125 group(all P < 0.01). The value of AI, W/D, IAR showed significantly lower than those in I/R group (all P <0.01). Meanwhile, light morphological and ultrastructure injury were found in SP600125 group.</p><p><b>CONCLUSION</b>SP600125 can suppress JNK signal pathway, up-regulate the ratio of Bcl-2/Bax to inhibit Caspase-3 dependent apoptosis, so that it protects lung tissue from ischemia/reperfusion injury.</p>


Subject(s)
Animals , Anthracenes , Pharmacology , Apoptosis , Caspase 3 , Metabolism , Lung , Metabolism , Pathology , MAP Kinase Signaling System , Phosphorylation , Proto-Oncogene Proteins c-bcl-2 , Metabolism , Rats , Rats, Wistar , Reperfusion Injury , Metabolism , Pathology , bcl-2-Associated X Protein , Metabolism
16.
Acta Physiologica Sinica ; (6): 609-616, 2012.
Article in English | WPRIM | ID: wpr-333164

ABSTRACT

NMDA-induced excitotoxicity cause severe neuronal damage including apoptosis and necrosis. The present study was aimed to evaluate the proportion of NMDA-induced apoptosis of rat cortical neurons and discover signal transduction mechanism. Caspase inhibitor and lactate dehydrogenase (LDH) assay were used to study the NMDA-induced apoptosis. To explore the involved signal pathways, the primary culture of rat cortical neurons were pretreated by the inhibitors of three MAPK pathways, extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p38 MAPK. With 2 h of NMDA treatment, cellular apoptosis was measured by caspase-3 activity, terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labeling (TUNEL) and Annexin V staining. The results showed that: (1) Caspase-dependent apoptosis accounted for 22.49% in NMDA-induced neuronal death; (2) Pretreatment with p38 MAPK inhibitor SB203580 (10 μmol/L) significantly decreased NMDA-mediated caspase-3 activity by 30.43% (P < 0.05). However, ERK inhibitor PD98059 (20 μmol/L) or JNK inhibitor SP600125 (20 μmol/L) did not influence caspase-3 activity; (3) Pretreatment with SB203580 significantly reduced the number of NMDA-induced TUNEL-positive cells by 33.10% (P < 0.05). PD98059 (20 μmol/L) or SP600125 (20 μmol/L) did not show obvious effect; (4) Pretreatment with SB203580 (10 μmol/L) significantly reduced the number of NMDA-induced early apoptotic neurons by 55.56% (P < 0.05). Also, SP600125 (20 μmol/L) significantly decreased the amount of late apoptotic/dead cells by 67.59% (P < 0.05). There was no effect of PD98059 (20 μmol/L). These results indicate that: (1) NMDA induces neuronal apoptosis besides necrosis; (2) p38 MAPK, but not JNK and ERK, is involved in NMDA-induced neuronal apoptosis, and inhibition of the apoptotic signaling pathway contributes to neuroprotection; (3) JNK activation might contribute to NMDA-induced neuronal necrosis rather than apoptosis.


Subject(s)
Animals , Anthracenes , Pharmacology , Apoptosis , Caspase 3 , Metabolism , Cells, Cultured , Extracellular Signal-Regulated MAP Kinases , Imidazoles , Pharmacology , JNK Mitogen-Activated Protein Kinases , MAP Kinase Signaling System , N-Methylaspartate , Pharmacology , Neurons , Cell Biology , Primary Cell Culture , Pyridines , Pharmacology , Rats , p38 Mitogen-Activated Protein Kinases
17.
Chinese Journal of Surgery ; (12): 166-170, 2012.
Article in Chinese | WPRIM | ID: wpr-257532

ABSTRACT

<p><b>OBJECTIVE</b>To study the effect and potential mechanism of expression of c-jun N-terminal kinase (JNK) signal pathway on neuron autophagy after diffuse brain injury (DBI).</p><p><b>METHODS</b>Male Sprague Dawley rats (n = 216) were randomly divided into four groups: DBI group (n = 54), SP600125 intervene group (n = 54), DMSO group (n = 54) and sham operation group (n = 54). DBI rat model was established according to the description of Marmarou DBI. At different time points (1, 6, 12, 24, 48 and 72 h) after operation, the histopathologic changes of neurons in cortex were observed by HE staining method; The expression of p-JNK, p-P53, DRAM and Beclin-1 were detected by Western blot and immunohistochemistry.</p><p><b>RESULTS</b>The results showed that under light microscope degenerated and necrotic neurons were observed to be scattered in cortex at 6 h after operation in DBI group, but these changes were low in SP600125 intervene group. Compared with SP600125 intervene group, the expression of p-JNK in DBI group were enhanced obviously at 6, 12 and 24 h (F = 17.902, P < 0.05); the expression of p-P53 in DBI group were enhanced obviously at 12, 24, 48 and 72 h (F = 7.107, P < 0.05); the expression of DRAM in DBI group were enhanced obviously at 6, 12, 24, 48 and 72 h (F = 15.455, P < 0.05); the expression of Beclin-1 in DBI group were enhanced obviously at 6, 12, 24, 48 and 72 h (F = 11.517, P < 0.05). Compared with DBI group, the expression of p-JNK, p-P53, DRAM and Beclin-1 in DMSO group were similar at 1, 6, 12, 24, 48 and 72 h (F = 1.509, P > 0.05).</p><p><b>CONCLUSIONS</b>The present results indicate that SP600125 can dramatically improve trauma brain injury from autophagy after DBI and the molecular mechanism is related to the modulation of JNK signal pathway following DBI, while it measures the neuron autophagy by means of intervening JNK signal pathway.</p>


Subject(s)
Animals , Anthracenes , Pharmacology , Autophagy , Brain Injuries , Metabolism , Pathology , Disease Models, Animal , JNK Mitogen-Activated Protein Kinases , Metabolism , Male , Neurons , Pathology , Rats , Rats, Sprague-Dawley
18.
Article in Chinese | WPRIM | ID: wpr-252554

ABSTRACT

<p><b>OBJECTIVE</b>To study and compare the therapeutic effects of electroacupuncture and probiotics combine Deanxit in treating diarrhea predominant irritable bowel syndrome (D-IBS).</p><p><b>METHODS</b>Totally 64 D-IBS patients accompanied with anxiety and/or depression were randomly assigned to the Western medicine group (Group A) and the electroacupuncture (EA) group (Group B), 30 patients in Group A and 34 patients in Group B. Patients in Group A took Bacillus licheniformis and Deanxit, while those in Group B received EA. Four weeks consisted of one therapeutic course. Three-month follow-up was carried out. The scoring for the digestive tract symptoms, HAMA score, and HAMD score were evaluated before and after treatment. The recurrence in the 3-month follow-up was also observed.</p><p><b>RESULTS</b>The total effect rate was 86.67% in Group A and 88.24% in Group B with no statistical difference between the two groups (P > 0.05). There was statistical difference in the scoring for the digestive tract symptoms, HAMA score, and HAMD score (P < 0.05, P < 0.01). There was no statistical difference in the improvement of defecation frequency score, HAMA score, HAMD score between the two groups after treatment (P > 0.05). Better effects on improving abdominal pain score and abdominal distention score was obtained in Group B (P < 0.01), while better effects on improving the stool form score and mucus score were obtained in Group A (P < 0.01). There was no statistical difference in the recurrence rate between the two groups within the two-month follow-up (P > 0.05). The recurrence rate within the 3-month follow-up was obviously lower in Group B than in Group A (P < 0.05).</p><p><b>CONCLUSIONS</b>EA and Western medicine (probiotics combined Deanxit) could effectively treat D-IBS patients accompanied with anxiety and/or depression. Both of them had different superiorities in improving symptoms. But EA had better long-term therapeutic effects.</p>


Subject(s)
Adult , Anthracenes , Therapeutic Uses , Diarrhea , Therapeutics , Drug Combinations , Electroacupuncture , Female , Flupenthixol , Therapeutic Uses , Humans , Irritable Bowel Syndrome , Therapeutics , Male , Middle Aged , Probiotics , Therapeutic Uses , Treatment Outcome
19.
Article in English | WPRIM | ID: wpr-728303

ABSTRACT

In this study, we focused to identify whether eupatilin (5,7-dihydroxy-3',4',6-trimethoxyflavone), an extract from Artemisia argyi folium, prevents H2O2-induced injury of cultured feline esophageal epithelial cells. Cell viability was measured by the conventional MTT reduction assay. Western blot analysis was performed to investigate the expression of 5-lipoxygenase by H2O2 treatment in the absence and presence of inhibitors. When cells were exposed to 600 microM H2O2 for 24 hours, cell viability was decreased to 40%. However, when cells were pretreated with 25~150 microM eupatilin for 12 hours, viability was significantly restored in a concentration-dependent manner. H2O2-treated cells were shown to express 5-lipoxygenase, whereas the cells pretreated with eupatilin exhibited reduction in the expression of 5-lipoxygenase. The H2O2-induced increase of 5-lipoxygenase expression was prevented by SB202190, SP600125, or NAC. We further demonstrated that the level of leukotriene B4 (LTB4) was also reduced by eupatilin, SB202190, SP600125, NAC, or nordihydroguaiaretic acid (a lipoxygenase inhibitor) pretreatment. H2O2 induced the activation of p38MAPK and JNK, this activation was inhibited by eupatilin. These results indicate that eupatilin may reduce H2O2-induced cytotoxicity, and 5-lipoxygenase expression and LTB4 production by controlling the p38 MAPK and JNK signaling pathways through antioxidative action in feline esophageal epithelial cells.


Subject(s)
Anthracenes , Arachidonate 5-Lipoxygenase , Artemisia , Blotting, Western , Cell Survival , Epithelial Cells , Flavonoids , Hydrogen , Hydrogen Peroxide , Imidazoles , Leukotriene B4 , Lipoxygenase , MAP Kinase Signaling System , Masoprocol , p38 Mitogen-Activated Protein Kinases , Pyridines
20.
Pesticidas ; 21: 117-128, jan.-out. 2011. graf, tab
Article in Portuguese | LILACS | ID: lil-671120

ABSTRACT

O objetivo deste estudo foi detectar a presença de antraceno,naftaleno, benzo[e]pireno e dibenzo[a,h]antraceno na fuligem dacana-de-açúcar e avaliar a toxicidade in vivo por meio da frequênciamédia de aberrações cromossômicas. A análise cromatográfi ca(em fase gasosa) efetuada na fuligem revelou a presença doshidrocarbonetos policíclicos aromáticos (HPA) estudados. O testede micronúcleo, realizado em células ósseas de camundongos,mostrou que benzo[e]pireno e dibenzo[a,h]antraceno apareceramcom frequência aumentada no micronúcleo nas doses testadas.No entanto, o antraceno e o naftaleno não apresentaram potencialgenotóxico. Os dados sugerem que é necessário considerar opotencial de risco oferecido pela fuligem para os cortadores de canae para a população que vive em áreas canavieiras.


Subject(s)
Anthracenes , Chromosome Aberrations , Polycyclic Aromatic Hydrocarbons , Mutation , Naphthalenes , Saccharum , Soot
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