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1.
Article in English | WPRIM | ID: wpr-921358

ABSTRACT

To explore interleukin-6 (IL-6) production and characterize lipid accumulation in L02 hepatocytes induced by sodium oleate. L02 hepatocytes were incubated with 0, 37.5, 75, 150, 300, 600, or 1,200 μmol/L sodium oleate for 24 h, and the supernatant was collected to detect the concentration of IL-6. L02 hepatocytes were incubated with 300, 150, 75, or 0 μmol/L sodium oleate for 0-24 h. The supernatant was collected for detection of IL-6 and free fatty acids. L02 hepatocytes treated with 300 μmol/L sodium oleate for 0-24 h were stained with Oil Red O. With extended sodium oleate incubation time, IL-6 levels increased, and free fatty acids decreased. After 24 h incubation, IL-6 levels increased as sodium oleate increased from 37.5 to 300 μmol/L (


Subject(s)
Dose-Response Relationship, Drug , Hepatocytes/metabolism , Humans , Interleukin-6/metabolism , Lipid Metabolism , Oleic Acid/administration & dosage , Time Factors
2.
Biol. Res ; 51: 17, 2018. graf
Article in English | LILACS | ID: biblio-950903

ABSTRACT

BACKGROUND: Improper control on reactive oxygen species (ROS) elimination process and formation of free radicals causes tissue dysfunction. Pineal hormone melatonin is considered a potent regulator of such oxidative damage in different vertebrates. Aim of the current communication is to evaluate the levels of oxidative stress and ROS induced damage, and amelioration of oxidative status through melatonin induced activation of signaling pathways. Hepatocytes were isolated from adult Labeo rohita and exposed to H2O2 at three different doses (12.5, 25 and 50 µM) to observe peroxide induced damage in fish hepatocytes. Melatonin (25, 50 and 100 µg/ml) was administered against the highest dose of H2O2. Enzymatic and non-enzymatic antioxidants such as malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT) and glutathione (GSH) was measured spectrophotometrically. Expression level of heat shock proteins (HSP70 and HSP90), HSPs-associated signaling molecules (Akt, ERK, cytosolic and nuclear NFkB), and melatonin receptor was also measured by western blotting analysis. RESULTS: H2O2 induced oxidative stress significantly altered (P < 0.05) MDA and GSH level, SOD and CAT activity, and up regulated HSP70 and HSP90 expression in carp hepatocytes. Signaling proteins exhibited differential modulation as revealed from their expression patterns in H2O2-exposed fish hepatocytes, in comparison with control hepatocytes. Melatonin treatment of H2O2-stressed fish hepatocytes restored basal cellular oxidative status in a dose dependent manner. Melatonin was observed to be inducer of signaling process by modulation of signaling molecules and melatonin receptor. CONCLUSIONS: The results suggest that exogenous melatonin at the concentration of 100 µg/ml is required to improve oxidative status of the H2O2-stressed fish hepatocytes. In H2O2 exposed hepatocytes, melatonin modulates expression of HSP70 and HSP90 that enable the hepatocytes to become stress tolerant and survive by altering the actions of ERK, Akt, cytosolic and nuclear NFkB in the signal transduction pathways. Study also confirms that melatonin could act through melatonin receptor coupled to ERK/Akt signaling pathways. This understanding of the mechanism by which melatonin regulates oxidative status in the stressed hepatocytes may initiate the development of novel strategies for hepatic disease therapy in future.


Subject(s)
Animals , Signal Transduction/drug effects , Oxidative Stress/drug effects , Hepatocytes/drug effects , Hydrogen Peroxide/pharmacology , Melatonin/pharmacology , Spectrophotometry , Superoxide Dismutase/drug effects , Catalase/drug effects , Catalase/metabolism , Blotting, Western , NF-kappa B/drug effects , NF-kappa B/metabolism , Reactive Oxygen Species/metabolism , MAP Kinase Signaling System/drug effects , Hepatocytes/metabolism , Proto-Oncogene Proteins c-akt/drug effects , Fishes , Glutathione/drug effects , Glutathione/metabolism , Malondialdehyde/metabolism
3.
Einstein (Säo Paulo) ; 15(2): 178-185, Apr.-June 2017. tab, graf
Article in English | LILACS | ID: biblio-891376

ABSTRACT

ABSTRACT Objective To analyze the influence of portal vein ligation in hepatic regeneration by immunohistochemical criteria. Methods Ten pigs divided into two groups of five animals underwent hepatectomy in two stages, and the groups were differentiated by ligation or not of the left portal vein tributary, which is responsible for vascularization of the left lateral and medial lobes of the pig liver. Five days after the procedure, the animals underwent liver biopsies for further analysis of histological and immunohistochemical with marker Ki67. Results The group submitted to hepatectomy with vascular ligation showed an increase of approximately 4% of hepatocytes in regeneration status, as well as a greater presence of Kupffer and inflammatory cells as compared to control. Conclusion As a result of positive cell replication observed through the Ki67 marker, we can suspect that the ligation of a tributary of the portal vein associated with liver resection promoted a greater stimulus of liver regeneration when compared to liver resection alone.


RESUMO Objetivo Analisar a influência da ligadura da tributária da veia porta no estímulo regenerativo hepático por meio de critérios imuno-histoquímicos. Métodos Dez suínos, divididos em dois grupos de cinco animais, foram submetidos à hepatectomia em dois estágios, sendo que os grupos foram diferenciados pela ligadura ou não da tributária da veia porta, responsável pela vascularização dos lobos lateral e medial esquerdos do fígado do suíno. Cinco dias após o procedimento, os animais foram reabordados para retirada de amostras hepáticas para posterior análise de histológica e imunoistoquímica com o marcador Ki67. Resultados O grupo submetido à hepatectomia com ligadura vascular apresentou incremento de 4% aproximadamente de hepatócitos em processo de regeneração, bem como grande número de células de Kupffer e células inflamatórias, quando comparado ao controle. Conclusão Em virtude da análise positiva da replicação celular observada por meio do marcador Ki67, pode-se observar que a ligadura de uma tributária da veia porta promoveu um maior estímulo de regeneração hepática, efeito observado com menor intensidade no grupo submetido apenas à ressecção hepática.


Subject(s)
Animals , Portal Vein/surgery , Parenchymal Tissue/surgery , Hepatectomy/methods , Liver/surgery , Liver Regeneration , Swine , Random Allocation , Ki-67 Antigen/metabolism , Hepatocytes/metabolism , Models, Animal , Parenchymal Tissue/pathology , Leukocytes , Ligation/methods , Liver/pathology
4.
Ann. hepatol ; 16(1): 48-56, Jan.-Feb. 2017. graf
Article in English | LILACS | ID: biblio-838085

ABSTRACT

Abstract: Liver fibrosis resulting from chronic liver injury are major causes of morbidity and mortality worldwide. Among causes of hepatic fibrosis, viral infection is most common (hepatitis B and C). In addition, obesity rates worldwide have accelerated the risk of liver injury due to nonalcoholic fatty liver disease (NAFLD) and nonalcoholic steatohepatitis (NASH). Also liver fibrosis is associated with the consumption of alcohol, or autoimmune hepatitis and chronic cholangiophaties. The response of hepatocytes to inflammation plays a decisive role in the physiopathology of hepatic fibrosis, which involves the recruitment of both pro- and anti-inflammatory cells such as monocytes and macrophages. As well as the production of other cytokines and chemokines, which increase the stimulus of hepatic stellate cells by activating proinflammatory cells. The aim of this review is to identify the therapeutic options available for the treatment of the liver fibrosis, enabling the prevention of progression when is detected in time.


Subject(s)
Humans , Animals , Liver Cirrhosis/drug therapy , Anti-Inflammatory Agents/therapeutic use , Time Factors , Signal Transduction/drug effects , Cell Communication/drug effects , Cytokines/metabolism , Treatment Outcome , Inflammation Mediators/metabolism , Disease Progression , Hepatocytes/drug effects , Hepatocytes/metabolism , Hepatocytes/pathology , Hepatic Stellate Cells/drug effects , Hepatic Stellate Cells/metabolism , Hepatic Stellate Cells/pathology , Liver/drug effects , Liver/metabolism , Liver/pathology , Liver Cirrhosis/etiology , Liver Cirrhosis/metabolism , Liver Cirrhosis/mortality , Anti-Inflammatory Agents/adverse effects
5.
Biol. Res ; 50: 2, 2017. graf
Article in English | LILACS | ID: biblio-838963

ABSTRACT

BACKGROUND: Cancer cells exhibit elevated levels of glucose uptake and may obtain pre-formed, diet-derived fatty acids from the bloodstream to boost their rapid growth; they may also use nucleic acid from their microenvironment. The study of processing nucleic acid by cancer cells will help improve the understanding of the metabolism of cancer. DNA is commonly packaged into a viral or lipid particle to be transferred into cells; this process is called transfection in laboratory. Cancer cells are known for having gene mutations and the evolving ability of endocytosis. Their uptake of DNAs might be different from normal cells; they may take in DNAs directly from the environment. In this report, we studied the uptake of DNAs in cancer cells without a transfection reagent. METHODS: A group of DNA fragments were prepared with PCR and labeled with isotope phosphorous-32 to test their uptake by Huh 7 (liver cancer) and THLE3 (normal liver cells) after incubation overnight by counting radioactivity of the cells' genomic DNA. Multiple cell lines including breast cancer and lung cancer were tested with the same method. DNA molecules were also labeled with fluorescence to test the location in the cells using a kit of "label it fluorescence in situ hybridization (FISH)" from Mirus (USA). RESULTS: The data demonstrated that hepatocellular carcinoma cells possess the ability to take in large DNA fragments directly without a transfection reagent whereas normal liver cells cannot. Huh7 and MDA-MB231 cells displayed a significantly higher Rhodamine density in the cytoplasmic phagosomes and this suggests that the mechanism of uptake of large DNA by cancer cells is likely endocytosis. The efficacy of uptake is related to the DNA's size. Some cell lines of lung cancer and breast cancer also showed similar uptake of DNA. CONCLUSIONS: In the present study, we have revealed the evidence that some cancer cells, but not nontumorigenic cells, can take DNA fragments directly from the environment without the aid of the transfecting reagent.


Subject(s)
Humans , Female , DNA/metabolism , Transfection , Neoplasms/genetics , Breast Neoplasms/genetics , Breast Neoplasms/pathology , alpha-Fetoproteins/metabolism , Cell Line , Polymerase Chain Reaction , In Situ Hybridization, Fluorescence , Hepatocytes/metabolism , Genomics , Cell Line, Tumor , Endocytosis/genetics , DNA Fragmentation , Lipids/pharmacology , Liver Neoplasms/genetics , Liver Neoplasms/pathology , Lung Neoplasms/genetics , Lung Neoplasms/pathology , Neoplasms/pathology
6.
Acta cir. bras ; 31(9): 608-614, Sept. 2016. tab, graf
Article in English | LILACS | ID: lil-795998

ABSTRACT

ABSTRACT PURPOSE: To assess the effect of aqueous extract of Peumus Boldus (AEPB) on the liver proliferative response after parcial hepatectomy of 70% (PH) in rodents. METHODS: Twenty Wistar rats were divided in two groups: AEPB100 (whose rats received 100mg/Kg of AEPB, once a day, orally, in 4 days prior to the first surgical procedure) and Vehicle (whose rats were treated similarly with distilled water). Both groups underwent PH. After 24 hours the remaining livers were removed for studying the proliferation of hepatocytes by Ki-67 and 2mL of blood were collected for serological assessment: cholesterol, glucose, triglycerides, aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, and total, direct and indirect bilirubin. All data were analyzed by Gaussian distribution. Statistically significant differences between mean values were analyzed using T Student's test. Non-Gaussian data were analyzed using Mann-Whitney's test. RESULTS: The liver of all these rats presented positive staining of Ki-67, indicating liver proliferation. Laboratory results showed no significant difference in serum values between the analyzed groups. The analysis of Ki-67 was significantly more positive in AEPB100 group than in Vehicle group. CONCLUSION: Aqueous extract of Peumus Boldus acute administration exerts significant positive effect on liver regeneration after 24h in rats that underwent parcial hepatectomy, while maintaining unchanged hepatic function.


Subject(s)
Animals , Female , Plant Extracts/pharmacology , Hepatocytes/drug effects , Peumus/chemistry , Hepatectomy/methods , Liver/physiology , Liver Regeneration/drug effects , Rats, Wistar , Plant Leaves/chemistry , Hepatocytes/metabolism , Disease Models, Animal , Liver/drug effects
7.
Arq. bras. cardiol ; 104(6): 443-449, 06/2015. tab
Article in English | LILACS | ID: lil-750700

ABSTRACT

Background: The aging process promotes a progressive increase in chronic-degenerative diseases. The effect of these diseases on the functional capacity has been well recognized. Another health parameter concerns “quality of life related to health”. Among the elderly population, cardiovascular diseases stand out due to the epidemiological and clinical impact. Usually, these diseases have been associated with others. This set of problems may compromise both independence and quality of life in elderly patients who seek cardiologic treatment. These health parameters have not been well contemplated by cardiologists. Objective: Evaluating, among the elderly population with cardiovascular disease, which are the most relevant clinical determinants regarding dependence and quality of life. Methods: This group was randomly and consecutively selected and four questionnaires were applied: HAQ, SF-36, PRIME-MD e Mini Mental State. Results: The study included 1,020 elderly patients, 63.3% women. The group had been between 60 and 97 years-old (mean: 75.56 ± 6.62 years-old). 61.4% were independent or mild dependence. The quality of life total score was high (HAQ: 88.66 ± 2.68). 87.8% of patients had a SF-36 total score > 66. In the multivariate analysis, the association between diagnoses and high degrees of dependence was significant only for previous stroke (p = 0.014), obesity (p < 0.001), lack of physical activity (p = 0.016), osteoarthritis (p < 0.001), cognitive impairment (p < 0.001), and major depression (p < 0.001). Analyzing the quality of life, major depression and physical illness for depression was significantly associated with all domains of the SF-36. Conclusion: Among an elderly outpatient cardiology population, dependence and quality of life clinical determinants are not cardiovascular comorbidities, especially the depression. .


Fundamento: Com o envelhecimento, a prevalência de doenças crônico-degenerativas sofreu aumento progressivo. A repercussão dessas doenças sobre a capacidade funcional foi reconhecida. Outro parâmetro de saúde é a “qualidade de vida relacionada à saúde”. Na população idosa, as doenças cardiovasculares destacam-se pelo impacto epidemiológico e clínico. Elas, geralmente, vêm associadas a outras afecções. Esse conjunto de problemas pode comprometer a independência e a qualidade de vida do idoso que busca tratamento cardiológico. Objetivo: Avaliar, em uma população de idosos cardiopatas, quais são os determinantes clínicos mais relevantes de dependência e de qualidade de vida. Métodos: O grupo foi selecionado aleatória e consecutivamente, sendo aplicados quatro questionários: HAQ, SF-36, PRIME‑MD e Mini Exame do Estado Mental. Resultados: Incluiu-se 1020 idosos, 63,3% mulheres. O grupo tinha em média 75,56 ± 6,62 anos. 61,4% mostrou-se independente ou com dependência leve. O escore de qualidade de vida foi elevado (HAQ: 88,66 ± 2,68). 87,8% dos pacientes apresentou escore total do SF-36 ≥ 66. À análise multivariada, a associação entre os diagnósticos e graus elevados de dependência foi significante apenas para acidente vascular cerebral prévio (p = 0,014), obesidade (p < 0,001), sedentarismo (p = 0,016), osteoartrite (p < 0,001), déficit cognitivo (p < 0,001), e depressão maior (p < 0,001). Ao analisarmos a qualidade de vida, a depressão maior e a depressão por doença física associou-se significativamente com todos os domínios do SF-36. Conclusão: Em uma população de idosos cardiopatas, os determinantes clínicos mais relevantes de prejuízos para dependência e qualidade de vida foram as comorbidades não cardiovasculares, particularmente a depressão. .


Subject(s)
Humans , Hepatocytes/pathology , Liver Regeneration , Liver Failure, Acute/metabolism , Apoptosis , /physiology , Fas Ligand Protein/physiology , Hepatocytes/metabolism , Liver Failure, Acute/therapy , Necrosis , Receptors, Tumor Necrosis Factor/metabolism , Signal Transduction , TNF-Related Apoptosis-Inducing Ligand/physiology , Tumor Necrosis Factor-alpha/metabolism
8.
Clinics ; 70(5): 373-379, 05/2015. tab, graf
Article in English | LILACS | ID: lil-748273

ABSTRACT

OBJECTIVE: Intestinal ischemia-reperfusion injury occurs in several clinical conditions and after intestinal transplantation. The aim of the present study was to investigate the phenomena of apoptosis and cell proliferation in a previously described intestinal ischemia-reperfusion injury autograft model using immunohistochemical markers. The molecular mechanisms involved in ischemia-reperfusion injury repair were also investigated by measuring the expression of the early activation genes c-fos and c-jun, which induce apoptosis and cell proliferation. MATERIALS AND METHODS: Thirty adult male Wistar rats were subjected to surgery for a previously described ischemia-reperfusion model that preserved the small intestine, the cecum and the ascending colon. Following reperfusion, the cecum was harvested at different time points as a representative segment of the intestine. The rats were allocated to the following four subgroups according to the reperfusion time: subgroup 1: 5 min; subgroup 2: 15 min; subgroup 3: 30 min; and subgroup 4: 60 min. A control group of cecum samples was also collected. The expression of c-fos, c-jun and immunohistochemical markers of cell proliferation and apoptosis (Ki67 and TUNEL, respectively) was studied. RESULTS: The expression of both c-fos and c-jun in the cecum was increased beginning at 5 min after ischemia-reperfusion compared with the control. The expression of c-fos began to increase at 5 min, peaked at 30 min, and exhibited a declining tendency at 60 min after reperfusion. A progressive increase in c-jun expression was observed. Immunohistochemical analyses confirmed these observations. CONCLUSION: The early activation of the c-fos and c-jun genes occurred after intestinal ischemia-reperfusion injury, and these genes can act together to trigger cell proliferation and apoptosis. .


Subject(s)
Animals , Mice , Rats , Endoplasmic Reticulum Stress , Fatty Acids/metabolism , Hepatocytes/physiology , Unfolded Protein Response , Acetylcysteine/metabolism , Cell Line, Tumor , Cells, Cultured , Glutathione/metabolism , Hepatocytes/metabolism , Oxidation-Reduction , Protein Folding
9.
Int. braz. j. urol ; 41(1): 116-123, jan-feb/2015. tab, graf
Article in English | LILACS | ID: lil-742880

ABSTRACT

Purpose To determine the safety of continued administration of antithrombotic agents during transperineal (TP) prostate biopsy. Patients and Methods A total of 811 men who underwent transrectal ultrasound (TRUS)-guided TP biopsy from January 2008 to June 2012 at our two institutions were retrospectively analyzed. Among these 811 men, 672 received no antithrombotic agents (group I), 103 received and continued administration of antithrombotic agents (group II), and 36 interrupted administration of antithrombotic agents (group III). Overall complications were graded and hemorrhagic complications were compared (group I with group II) using propensity score matching (PSM) analysis. Results An overall complication rate of 4.6% was recorded. Hemorrhagic complications occurred in 1.8% and they were virtually identical in all the three groups, and no severe hemorrhagic complications occurred. One patient in group III required intensive care unit admission for cerebral infarction. PSM analysis revealed no statistical difference between groups I and II with regard to the incidence of gross hematuria, perineal hematoma, and rectal bleeding. Multiple regression analysis revealed that hemorrhagic complications were associated with lower body mass index (<21 kg/m2, P=0.0058), but not with administration of antithrombotic agents. Conclusions Continued administration of antithrombotic agents does not increase the risk of hemorrhagic complications; these agents are well tolerated during TP biopsy. .


Subject(s)
Adolescent , Adult , Animals , Child , Child, Preschool , Female , Humans , Infant , Male , Mice , Young Adult , Hepatocytes/transplantation , Liver Diseases/pathology , Liver Transplantation/methods , Liver/pathology , Hepatocytes/cytology , Hepatocytes/metabolism , Liver Diseases/metabolism , Liver Diseases/surgery , Liver/metabolism , Liver/surgery
10.
Biol. Res ; 48: 1-8, 2015. graf
Article in English | LILACS | ID: biblio-950787

ABSTRACT

BACKGROUND: Hepcidin, encoding by HAMP gene, is the pivotal regulator of iron metabolism, controlling the systemic absorption and transportation of irons from intracellular stores. Abnormal levels of HAMP expression alter plasma iron parameters and lead to iron metabolism disorders. Therefore,itis animportant goal to understand the mechanisms controlling HAMP gene expression. RESULTS: Overexpression of Sox2 decrease basal expression of HAMP or induced by IL-6 or BMP-2, whereas, knockdown of Sox2 can increase HAMP expression, furthermore, two potential Sox2-binding sites were identified within the human HAMP promoter. Indeed, luciferase experiments demonstrated that deletion of any Sox2-binding site impaired the negative regulation of Sox2 on HAMP promoter transcriptional activity in basal conditions. ChIP experiments showed that Sox2 could directly bind to these sites. Finally, we verified the role of Sox2 to negatively regulate HAMP expression in human primary hepatocytes. CONCLUSION: We found that Sox2 as a novel factor to bind with HAMP promoter to negatively regulate HAMP expression, which may be further implicated as a therapeutic option for the amelioration of HAMP-overexpression-related diseases, including iron deficiency anemia.


Subject(s)
Humans , Gene Expression Regulation, Neoplastic/genetics , Hepatocytes/metabolism , SOXB1 Transcription Factors/genetics , Hepcidins/genetics , Plasmids/genetics , Binding Sites , Interleukin-6/metabolism , Promoter Regions, Genetic/genetics , Bone Morphogenetic Protein 2/metabolism , SOXB1 Transcription Factors/metabolism , Gene Knockdown Techniques , Hep G2 Cells , Hepcidins/metabolism , Genetic Vectors , Anemia/genetics , Anemia/metabolism , Iron/metabolism , Luciferases
11.
Article in English | WPRIM | ID: wpr-157210

ABSTRACT

Hepatitis B virus (HBV) is the prototype of hepatotropic DNA viruses (hepadnaviruses) infecting a wide range of human and non-human hosts. Previous studies with duck hepatitis B virus (DHBV) identified duck carboxypeptidase D (dCPD) as a host specific binding partner for full-length large envelope protein, and p120 as a binding partner for several truncated versions of the large envelope protein. p120 is the P protein of duck glycine decarboxylase (dGLDC) with restricted expression in DHBV infectible tissues. Several lines of evidence suggest the importance of dCPD, and especially p120, in productive DHBV infection, although neither dCPD nor p120 cDNA could confer susceptibility to DHBV infection in any cell line. Recently, sodium taurocholate cotransporting polypeptide (NTCP) has been identified as a binding partner for the N-terminus of HBV large envelope protein. Importantly, knock down and reconstitution experiments unequivocally demonstrated that NTCP is both necessary and sufficient for in vitro infection by HBV and hepatitis delta virus (HDV), an RNA virus using HBV envelope proteins for its transmission. What remains unclear is whether NTCP is the major HBV receptor in vivo. The fact that some HBV patients are homozygous with an NTCP mutation known to abolish its receptor function suggests the existence of NTCP-independent pathways of HBV entry. Also, NTCP very likely mediates just one step of the HBV entry process, with additional co-factors for productive HBV infection still to be discovered. NTCP offers a novel therapeutic target for the control of chronic HBV infection.


Subject(s)
Animals , Carboxypeptidases/genetics , Gene Products, pol/genetics , Heparan Sulfate Proteoglycans/metabolism , Hepatitis B virus/physiology , Hepatocytes/metabolism , Organic Anion Transporters, Sodium-Dependent/antagonists & inhibitors , RNA Interference , Symporters/antagonists & inhibitors , Viral Envelope Proteins/metabolism , Virus Internalization
12.
Braz. dent. j ; 25(5): 372-378, Sep-Oct/2014. tab, graf
Article in English | LILACS | ID: lil-731053

ABSTRACT

This study aimed to evaluate the immunoexpression of glucose transporters 1 (GLUT-1) and 3 (GLUT-3) in metastatic and non-metastatic lower lip squamous cell carcinoma (LLSCC). Twenty LLSCCs with regional nodal metastasis and 20 LLSCCs without metastasis were selected. The distribution of staining and the percentage of GLUT-1 and GLUT-3 staining in each tumor core and at the deep invasive front were assessed. Most tumors (70%) exhibited peripheral staining for GLUT-1 in nests, sheets and islands of neoplastic cells, whereas predominantly central staining was observed for GLUT-3 (72.5%). A high percentage of GLUT-1-positive cells was observed at the deep invasive front and in the tumor core of metastatic and non-metastatic tumors (p>0.05). The percentage of GLUT-1-positive cells was much higher than that of GLUT-3-positive cells both in the deep invasive front (p<0.001) and in the tumor core (p<0.001) of LLSCCs. No significant differences in the percentage of GLUT-1- and GLUT-3-positive cells were observed according to nodal metastasis, clinical stage or histological grade of malignancy (p>0.05). In conclusion, the results of the present study suggest an important role of GLUT-1 in glucose uptake in LLSCCs, although this protein does not seem to be involved in the progression of these tumors. On the other hand, GLUT-3 expression may represent a secondary glucose uptake mechanism in LLSCCs.


Este estudo objetivou avaliar a imunoexpressão dos transportadores de glicose 1 (GLUT-1) e 3 (GLUT-3) em carcinomas de células escamosas de lábio inferior (CCELI) metastáticos e não-metastáticos. Vinte CCELIs com metástase nodal regional e 20 CCELI sem metástase foram selecionados. Foram analisados a distribuição da imunomarcação e o percentual de imunorreatividade para GLUT-1 e GLUT-3 no centro tumoral e no front de invasão tumoral. A maioria dos tumores (70%) revelou marcação para GLUT-1 em áreas periféricas dos ninhos, lençóis e ilhas de células neoplásicas, ao passo que GLUT-3 revelou predomínio de marcação em áreas centrais (72.5%). Um alto percentual de células positivas para GLUT-1 foi observado no front de invasão e no centro tumoral das lesões metastáticas e não-metastáticas (p>0,05). O percentual de células positivas para GLUT-1 foi superior ao percentual de células positivas para GLUT-3, tanto no front de invasão (p<0,001) quanto no centro tumoral (p<0,001) dos CCELIs. Não foram observadas diferenças significativas no percentual de células positivas para GLUT-1 e GLUT-3 em relação à mestástase nodal, ao estádio clínico ou ao grau histológico de malignidade (p>0,05). Em conclusão, os resultados do presente estudo sugerem um importante papel para GLUT-1 na absorção de glicose nos CCELIs, embora esta proteína não pareça estar envolvida na progressão destes tumores. Por outro lado, a expressão de GLUT-3 pode representar um mecanismo secundário para a absorção de glicose nos CCELIs.


Subject(s)
Animals , Female , Male , Rats , Estradiol/pharmacology , Hepatocytes/drug effects , Oxidative Stress , Apoptosis/physiology , bcl-X Protein , Cells, Cultured , Flow Cytometry , Hepatocytes/metabolism , L-Lactate Dehydrogenase/metabolism , Lipid Peroxidation , /metabolism , Rats, Wistar , Thiobarbituric Acid Reactive Substances/metabolism
13.
Rev. bras. enferm ; 67(5): 700-707, Sep-Oct/2014. tab, graf
Article in Portuguese | LILACS, BDENF | ID: lil-731203

ABSTRACT

O artigo descreve a construção de um questionário para avaliação da qualidade do atendimento de um Time de Resposta Rápida, em Hospital Universitário de Londrina-PR, fundamentado no modelo conceitual de Donabedian (estrutura-processoresultado). A coleta de dados ocorreu no mês de março de 2012 e o processo de adequação do questionário foi desenvolvido com a aplicação da Técnica Delphi com a participação de 15 especialistas. Ao término do estudo obteve-se um questionário com 37 enunciados, sendo alcançado índice de concordância final na pesquisa com valores superiores a 80% para todos os conceitos. Espera-se que as contribuições do grupo de especialistas tornem o instrumento confiável e seja aplicado em outros serviços semelhantes. Aplicações futuras deste instrumento poderão trazer subsídios para melhor avaliação da qualidade dos serviços de equipes de Resposta Rápida.


The paper describes the construction of a questionnaire to assess the quality of care of a Rapid Response Team at the University Hospital of Londrina, based on the conceptual model of Donabedian (structure-process-outcome). Data collection occurred in March 2012 and the process of adjusting the questionnaire was developed with the application of the Delphi technique involving 15 experts. At the end of the study the questionnaire contained 37 statements, achieving final compliance level higher than 80% in all concepts. It is hoped that the contributions of the expert group produce a more reliable questionnaire to be applied in other similar services. Future applications of this instrument may provide information to better assess the quality of teams of Rapid Response services.


El artículo describe la construcción de un cuestionario para la evaluación de la calidad de la atención de un Equipo de Respuesta Rápida en un Hospital Universitario de Londrina, Paraná, basado en el modelo conceptual de Donabedian (estructura-procesoresultado). La recolección de datos ocurrió durante el mes de marzo de 2012 y el proceso de ajuste del cuestionario fue desarrollado por medio de la Técnica Delphi con la participación de 15 especialistas. Al término del estudio se obtuvo un cuestionario con 37 enunciados, alcanzándose un índice de concordancia final en la investigación con valores superiores al 80% para todos los conceptos. Se espera que las contribuciones del grupo de especialistas afiancen la confiabilidad del instrumento y que el cuestionario sea utilizado en otros servicios semejantes. Las aplicaciones futuras podrán traer subsidios para evaluar mejor la calidad de los servicios de los equipos de Respuesta Rápida.


Subject(s)
Humans , Animals , Male , Rats , Alcoholic Beverages/toxicity , Hepatocytes/drug effects , Metallothionein/biosynthesis , Cell Division/drug effects , Collagen/biosynthesis , Hepatocytes/metabolism , Hepatocytes/pathology , Liver Cirrhosis, Alcoholic/etiology , Liver Cirrhosis, Alcoholic/prevention & control , Rats, Sprague-Dawley
14.
Biocell ; 36(2): 63-71, Aug. 2012. graf, tab
Article in English | LILACS | ID: lil-662143

ABSTRACT

The flower of Butea monosperma (Lam.) (Fabaceae) has been used in traditional Indian medicine in the treatment of many ailments including liver disorders. To understand the pharmacological basis of its beneficial effects, the extracts of dried flowers in water, methanol, butanol, ethyl acetate and acetone were evaluated for free radical scavenging and pro-apoptotic activities in cell cultures (human hepatoma Huh-7 cell line and immortalized AML-12 mouse hepatocytes). Butrin and butein -the active constituents of flower extracts- were used as reference molecules. The levels of cell injury markers like lactate dehydrogenase, glutathione and lipid peroxidation and primary antioxidant enzymes glutathione S-transferase and catalase were also measured. The aqueous and butanolic extracts exhibited better 2,2-diphenyl-1-picrylhydrazyl scavenging and cytotoxic activities in hepatoma cells than in immortalized hepatocytes. Interestingly, butein inhibited 2,2-diphenyl-1-picrylhydrazyl radical better than butrin. The aqueous and butanolic extracts were further investigated for hepatoprotection against carbon tertrachloride-induced biochemical changes and cell death. Both extracts, just as butrin and butein, significantly reversed the cellular glutathione levels and lipid peroxidation, and glutathione-S-transferase activity. Lactate dehydrogenase leakage and cell death were also prevented. However, only butein revived the catalase activity. Thus, the butein content of Butea monosperma flower extracts is important for free radical scavenging activity, apoptotic cell death and protection against oxidative injury in hepatic cells.


Subject(s)
Animals , Humans , Mice , Antioxidants/pharmacology , Apoptosis/drug effects , Butea/chemistry , Chalcones/pharmacology , Flowers/chemistry , Free Radical Scavengers/pharmacology , Free Radicals/metabolism , Plant Extracts/pharmacology , Antioxidants/isolation & purification , Cells, Cultured , Carcinoma, Hepatocellular/drug therapy , Carcinoma, Hepatocellular/metabolism , Carcinoma, Hepatocellular/pathology , Chalcones/isolation & purification , Flavonoids/isolation & purification , Flavonoids/pharmacology , Free Radical Scavengers/isolation & purification , Glutathione Transferase/metabolism , Glutathione/metabolism , Hepatocytes/cytology , Hepatocytes/drug effects , Hepatocytes/metabolism , Lipid Peroxidation/drug effects , Liver Neoplasms/drug therapy , Liver Neoplasms/metabolism , Liver Neoplasms/pathology , Oxidation-Reduction
15.
Braz. j. med. biol. res ; 45(6): 482-487, June 2012. ilus, tab
Article in English | LILACS | ID: lil-622776

ABSTRACT

This study explored the reduction of adenosine triphosphate (ATP) levels in L-02 hepatocytes by hexavalent chromium (Cr(VI)) using chi-square analysis. Cells were treated with 2, 4, 8, 16, or 32 μM Cr(VI) for 12, 24, or 36 h. Methyl thiazolyl tetrazolium (MTT) experiments and measurements of intracellular ATP levels were performed by spectrophotometry or bioluminescence assays following Cr(VI) treatment. The chi-square test was used to determine the difference between cell survival rate and ATP levels. For the chi-square analysis, the results of the MTT or ATP experiments were transformed into a relative ratio with respect to the control (%). The relative ATP levels increased at 12 h, decreased at 24 h, and increased slightly again at 36 h following 4, 8, 16, 32 μM Cr(VI) treatment, corresponding to a "V-shaped" curve. Furthermore, the results of the chi-square analysis demonstrated a significant difference of the ATP level in the 32-μM Cr(VI) group (P < 0.05). The results suggest that the chi-square test can be applied to analyze the interference effects of Cr(VI) on ATP levels in L-02 hepatocytes. The decreased ATP levels at 24 h indicated disruption of mitochondrial energy metabolism and the slight increase of ATP levels at 36 h indicated partial recovery of mitochondrial function or activated glycolysis in L-02 hepatocytes.


Subject(s)
Animals , Humans , Adenosine Triphosphate/metabolism , Carcinogens, Environmental/toxicity , Chromium/toxicity , Hepatocytes/drug effects , Analysis of Variance , Adenosine Triphosphate/chemistry , Cell Culture Techniques , Chi-Square Distribution , China , Coloring Agents , Cell Survival/drug effects , Hepatocytes/metabolism , Mitochondria, Liver/metabolism , Tetrazolium Salts , Thiazoles
16.
Rev. salud pública ; 16(3): 347-360, 2012. ilus, tab
Article in Spanish | LILACS | ID: lil-729657

ABSTRACT

Objectivo El propósito del presente estudio es analizar las inequidades socioeconómicas en la utilización de servicios de salud en el Ecuador, las inequidades en la distribución geográfica de recursos humanos en salud, y reflexionar sobre los retos de equidad que el sistema de salud ecuatoriano enfrenta en la actualidad. Métodos Se utilizó la Encuesta Demográfica y de Salud Materno-Infantil (ENDEMAIN 2004) como la principal fuente de datos, cuya muestra es representativa de la población ecuatoriana. Para estimar los efectos en utilización de servicios de salud utilizamos análisis multivariado multinivel (usando el paquete estadístico MLWiN 2.02) y análisis espacial de recursos en salud (usando GeoDa 1.0.1 ). Resultados Nuestro análisis encontró que inequidades sociales, económicas y geográficas limitan el acceso a servicios de salud en el Ecuador. Hogares de bajos recursos, indígenas y aquellos que viven enáreas rurales (muchos con las tres características a la vez) tienen menos posibilidades de utilizar servicios de salud. A pesar de la marcada concentración de proveedores de salud en zonas urbanas, encontramos que la presencia de personal de salud (excluyendo a médicos) en entidades públicas rurales incrementa la posibilidad de utilización de servicios preventivos y curativos. Conclusiones Los esfuerzos para transformar el sistema de salud deben reducir barreras sociales, culturales, financieras; y las desigualdades en la distribución de recursos humanos en salud, particularmente en elárea rural. Consideramos que la orientación comunitaria y familiar de los servicios, y el incremento de espacios de participación ciudadana son necesarios para reducir dichas inequidades.


Objective The present study was aimed at analysing socioeconomic inequity regarding the use of health services in Ecuador, inequity regarding the geographic distribution of healthcare-related human resources and reflecting on the challenges concerning equity which the Ecuadorian health system is currently facing. Methods The Ecuadorian Demographic, Maternal and Infant Health Survey (2004) was used as the main data source, as its sample was representative of the Ecuadorian population. Multilevel multivariate analysis (MLWiN 2.02 statistical software) and spatial data analysis regarding health resources (GeoDa 1.0.1) were used for estimating the effects of using health services. Results It was found that social, economic and geographic inequity limited access to health services in Ecuador. People living in low economic resource households or indigenous housing and people living in rural areas (many of them having all three characteristics at the same time) had less possibility of using health services. In spite of a marked concentration of health-service providers in urban areas, it was found that the presence of healthcare personnel (excluding doctors) in rural public entities increased the possibility of using preventative and curative services. Conclusions Efforts at transforming the Ecuadorian health system must be aimed at reducing social, cultural and financial barriers and inequality regarding the distribution de healthcare-related human resources, particularly in rural areas. Community and family orientation of the services and increasing spaces for citizen participation are necessary for reducing such inequity.


Subject(s)
Animals , Rats , Collagen/metabolism , Hepatocytes/drug effects , Iron/pharmacology , /metabolism , Actins/metabolism , Cell Division/drug effects , Collagen/drug effects , Ferric Compounds/pharmacology , Gene Expression/drug effects , Hepatocytes/metabolism , Malondialdehyde/metabolism , Rats, Sprague-Dawley
17.
Archives of Iranian Medicine. 2011; 14 (4): 244-249
in English | IMEMR | ID: emr-129710

ABSTRACT

Orthotropic liver transplantation [OLT] is the final procedure of both end stage and metabolic liver diseases. Hepatocyte transplantation is an alternative for OLT, but the sources of hepatocytes are limited. Bone marrow mesenchymal stem cells [BM-MSCs] can differentiate into hepatocyte-like cells and are a potential alternative source for hepatocytes. We aimed to investigate the differentiation potential of BM-MSCs into hepatocyte-like cells. Human BM-MSCs from a healthy donor were cultured and differentiated into hepatocyte-like cells. We investigated the expression of hepatocyte-specific markers in MSC-derived hepatocyte-like cells [MSC-HLC[s]] and evaluated their functionality using metabolic assays. MSC-HLCs expressed hepatocyte-specific markers at both mRNAand protein levels. In addition, the cells had the ability to uptake low density lipoprotein [LDL], clear ammonia, secrete albumin, and store glycogen. MSC-HLCs were transplanted into a familial hypercholesteromia patient. Human MSCs can be differentiated into partially functional hepatocyte-like cells. Thus, they could be a potential source for cell therapy in liver disorders


Subject(s)
Humans , Male , Adult , Bone Marrow Cells/cytology , Hepatocytes/metabolism , Mesenchymal Stem Cells/cytology , RNA, Messenger/metabolism , Bone Marrow Cells/physiology , Hepatocytes/transplantation , Mesenchymal Stem Cells/physiology , In Vitro Techniques , Receptors, LDL/genetics , Keratin-18/genetics , Keratin-19/genetics
18.
Braz. j. med. biol. res ; 43(1): 36-42, Jan. 2010. tab, ilus
Article in English | LILACS | ID: lil-535640

ABSTRACT

Transforming growth factor-â1 (TGF-â1) plays an important role in the fibrogenic process in the liver. The aim of the present study was to explore the action of TGF-â1 on fibronectin expression in rat hepatic stem-like cells and the underlying mechanisms. The level of fibronectin expression was determined in hepatic stem-like cells (WB cells) before and after TGF-â1 stimulation by RT-PCR and Western blot methods. Using immunogold transmission electron microscopy and the Western blot method, we observed the result of the expression and the distribution of cAMP, phosphorylated Smad3 and Smad7 before and after TGF-â1 treatment. The levels of fibronectin expression in both mRNA and protein increased 4- to 5-fold after TGF-â1 stimulation, reaching an optimum level after 8 h and then gradually falling back. Similarly, TGF-â1 stimulation resulted in an increase of cAMP in WB cells, peaking at 8 h. After treatment with TGF-â1 for 24 h, the expression of cAMP gradually decreased. In addition, we found that TGF-â1 treatment also contributed to the increased expression and to changes in cellular distribution of phosphorylated Smad3 (translocation from the cytoplasm to the nucleus) and Smad7 (translocation from the nucleus to the cytoplasm) in WB cells. The present study demonstrates that TGF-â is involved in the fibrogenic process in hepatic stem cells through up-regulation of fibronectin expression, and the mechanisms underlying this process may be associated with the activation of cAMP and Smad pathways.


Subject(s)
Animals , Rats , Fibronectins/metabolism , Hepatocytes/metabolism , Stem Cells/metabolism , Transforming Growth Factor beta/pharmacology , Blotting, Western , Cyclic AMP/metabolism , Fibronectins/genetics , Hepatocytes/pathology , Liver Cirrhosis/etiology , Microscopy, Electron, Transmission , Reverse Transcriptase Polymerase Chain Reaction , RNA, Messenger/metabolism , Smad Proteins/metabolism , Stem Cells/pathology
19.
Article in English | IMSEAR | ID: sea-16397

ABSTRACT

BACKGROUND & OBJECTIVES: It has been reported that some proteins are released from mitochondria during liver regeneration after partial hepatectomy (PH), but the relationship between proteins release and mitochondrial permeability transition (MPT) remains unclear. We undertook this study to demonstrate the changes of mitochondrial ultrastructure and proteins release during liver regeneration and to determine the relationship between proteins release and MPT in liver regeneration in rats. METHODS: After PH and administration of cyclosporin-A (CsA, a specific inhibitor of MPT), ultrastructural morphology of mitochondria in the remnant liver were determined by electron microscopy. Catalytic activity of mitochondrial and cytosolic proteins including aspartate aminotransferase (AST) and glutamic acid dehydrogenase (GDH) was measured. RESULTS: The liver mitochondria at 24 and 72 h were quite variable in morphology and ultrastructure. The enzyme activities of AST and GDH in cytosol released from mitochondrial matrix changed significantly at 24 and 72 h. CsA can inhibit the permeability of mitochondria partly at the same time. INTERPRETATION & CONCLUSIONS: The changes of mitochondria in ultrastructure reflected the feature of MPT, and the changes of enzymes activities released from mitochondrial matrix were consistent with those of mitochondrial ultrastructure. CsA can inhibit these changes to some extent. There was a close relationship of MPT with mitochondrial ultrastructure and proteins release during liver regeneration.


Subject(s)
Analysis of Variance , Animals , Aspartate Aminotransferase, Mitochondrial/metabolism , Cyclosporine , Hepatectomy , Hepatocytes/metabolism , Liver Regeneration/physiology , Male , Microscopy, Electron , Mitochondria/ultrastructure , Permeability , Rats
20.
Article in Korean | WPRIM | ID: wpr-28356

ABSTRACT

BACKGROUND/AIMS: It is essential to develop an in vitro culture model of primary hepatocytes for the study of hepatocellular function and the pathogenesis of hepatitis C virus (HCV) infection. In this study, we have established the immortalized primary human hepatocyte (IPHH) and performed in vitro culture of HCV derived from human patient. METHODS: Primary human hepatocytes were isolated from surgically resected liver tissue and then were immortalized by transfection with the SV40 large T antigen. The characterization of the IPHH during culture was analyzed by immunocytochemistry, RT-PCR, Western blot, ELISA, and soft agar assay. Next, sera and/or liver tissue homogenates from surgically resected liver tissues of patients with HCV infection were inoculated for the culture of HCV in IPHH. After HCV RNA extraction from IPHH and culture media, positive or negative stranded HCV RNA was examined by specific nest RT-PCR. RESULTS: IPHH expressed liver-associated proteins but did not express alpha-fetoprotein. Also IPHH showed ammonia removal activity. With regard to its malignant potential, colony formation in soft agar assay was not observed. Next, positive and negative stranded HCV RNAs in IPHH infected with patient's sera plus liver tissue homogenates were clearly detected whereas those in IPHH infected with only patient's sera were not detected. CONCLUSIONS: These results demonstrated the phenotypic characteristics of IPHH and the feasibility in vitro culture system of HCV infected human samples. This system might be useful for study of pathogenesis of HCV infection or hepatocyte-based applications.


Subject(s)
Antigens, Viral, Tumor/genetics , Base Sequence , Carcinogenicity Tests , Cell Culture Techniques , Cells, Cultured , Cells, Immobilized , Hepacivirus/isolation & purification , Hepatocytes/metabolism , Humans , Liver Function Tests , Models, Biological , RNA Probes , RNA, Viral/analysis , Reverse Transcriptase Polymerase Chain Reaction
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