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1.
Rev. chil. endocrinol. diabetes ; 14(1): 7-13, 2021. tab, ilus
Article in Spanish | LILACS | ID: biblio-1146465

ABSTRACT

INTRODUCCIÓN: La enfermedad del hígado graso no alcohólico (EHGNA) es la forma más común de enfermedad hepática. A nivel celular se caracteriza por la acumulación de triglicéridos (TG) en forma de gotas lipídicas (GL) dando lugar a esteatosis e inflamación. Entre los factores relevantes para la síntesis de TG se encuentran las enzimas DGAT1/2 que catalizan la etapa final de la síntesis de TG, y la proteína FABP4 que transporta lípidos intracelulares y se expresa en modelos de enfermedad hepática dependiente de obesidad. Por otra parte, TNF-α es una reconocida citoquina involucrada en el proceso inflamatorio en la EHGNA. La medicina popular del norte de Chile ha utilizado la planta Lampaya medicinalis Phil. (Verbenaceae) para el tratamiento de algunas enfermedades inflamatorias. OBJETIVO: Evaluar el efecto de un extracto hidroalcóholico de lampaya (EHL) sobre la esteatosis y expresión de marcadores de inflamación en hepatocitos tratados con ácidos grasos. Diseño experimental: Estudio in vitro en cultivos de la línea celular humana HepG2 tratadas con ácido oleico (AO) y ácido palmítico (AP). MÉTODOS: Se incubó hepatocitos HepG2 con AO/AP por 24 horas en presencia o no de EHL. Se evaluó la presencia de GL y el contenido de TG intracelulares por Oil Red O y Nile Red, respectivamente. La expresión de DGAT1/2, FABP4 y TNF-α fue evaluada por qPCR. RESULTADOS: Los hepatocitos tratados con AO/AP mostraron un aumento en las GL y TG, así como una mayor expresión de DGAT2 en comparación al control. El cotratamiento con EHL revirtió los efectos inducidos por AO/AP. CONCLUSIONES: EHL revierte el incremento en las GL, TG y en la expresión de DGAT2 inducido por AO/AP en células HepG2. Estos hallazgos sugieren un efecto hepatoprotector de la Lampaya contra la esteatosis, y apoyarían su uso complementario en el tratamiento de patologías con componente inflamatorio como la EHGNA.


Non-alcoholic fatty liver disease (NAFLD) is the most common liver disease. At the cellular level, it is characterized by the accumulation of triglycerides (TG) in the form of lipid droplets (LD), which leads to steatosis and inflammation. Among relevant factors for TG synthesis are the enzymes DGAT1/2 catalyzing the final stage of TG synthesis, and the protein FABP4 which transports intracellular lipids and is expressed in cell models of obesity-dependent liver disease. Additionally, TNF-α is a cytokine involved in the inflammatory process associated to NAFDL. Lampaya medicinalis Phil. (Verbenaceae) is a plant used in folk medicine in northern Chile to treat some inflammatory diseases. OBJECTIVE: To evaluate the effect of the hydroalcoholic extract of lampaya (HEL) on steatosis and the expression of inflammatory markers in hepatocytes treated with fatty acids. Study design: In vitro study in cultures of the human HepG2 cell line treated with oleic acid (OA) and palmitic acid (PA). METHODS: HepG2 hepatocytes were incubated with OA/PA for 24 hours in the presence and absence of HEL. The formation of LD and the accumulation of intracellular TG were assessed by Oil Red O and Nile Red, respectively. The expression of DGAT1/2, FABP4 and TNF-α was assessed by qPCR. RESULTS: The treatment with OA/PA increased the levels of LD and TG as well as the expression of DGAT2 in HepG2 hepatocytes compared to control cells. HEL cotreatment counteracted OA/PA-induced effects. CONCLUSIONS: HEL prevents the increase in LD and TG levels and DGAT2 expression induced by OA/PA in HepG2 cells. These findings suggest that lampaya may have a protective effect against hepatic steatosis, which would support its complementary use in the treatment of pathologies associated with inflammation, such as NAFLD.


Subject(s)
Humans , Plant Extracts/pharmacology , Hepatocytes/drug effects , Verbenaceae/chemistry , Non-alcoholic Fatty Liver Disease/drug therapy , Triglycerides/analysis , In Vitro Techniques , Plant Extracts/therapeutic use , Cell Survival , Polymerase Chain Reaction , Cell Culture Techniques , Oleic Acid , Ethanol/chemistry , Hep G2 Cells/drug effects , Inflammation
2.
Rev. chil. endocrinol. diabetes ; 11(2): 47-53, abr. 2018. tab, graf
Article in Spanish | LILACS | ID: biblio-914719

ABSTRACT

Introduction: The Calcium Sensing Receptor (CaSR) is expressed in human fat cells, and its stimulation may be associated with adipose tissue dysfunction. The multisystemic character of obesity and the search of deepening the scope of the activation of CaSR in this disorder allows us to study the response of this protein in tissues that differ from adipose. Objective: To evaluate the effect of CaSR activation on the expression of lipogenic genes in a model of excess glucose and fatty acids in HepG2 human liver cells. Materials and methods: The effect of the calcimimetic cinacalcet (allosteric agonist of CaSR) on the content of triglycerides (fluorimetry) in a model of glucose supply and on the expression of lipogenic genes (qPCR) in hyperglycemia and hyperlipidemia conditions in the Liver cell line HepG2. Results: Cinacalcet, glucose (25 mM) and oleic acid (0.6 mM) did not affect cell viability. Activation of CaSR in the presence of glucose failed to increase the intracellular triglyceride content at 72 hours. Under these conditions, no response was observed for the factors coding for lipogenic genes (SREBP1c and FAS) at 24 hours of stimulation with cinacalcet in the liver cells. In the case of the over supply of fatty acids, the HepG2 cells did not show a variation in the gene expression of the DGAT enzymes after exposure to cinacalcet. Conclusion:Under conditions of glucose exposure, cinacalcet did not show a response in the triglyceride content, nor in the expression of genes related to hepatic lipogenesis. Therefore, stimulation of CaSR would not be associated with hepatic steatosis in HepG2 cells exposed to glucose.


Subject(s)
Humans , Receptors, Calcium-Sensing , Lipogenesis , Hep G2 Cells , Cell Survival , Real-Time Polymerase Chain Reaction
3.
Biol. Res ; 41(2): 197-204, 2008. graf
Article in English | LILACS | ID: lil-495754

ABSTRACT

It has been proposed that differences in adipocyte function and/or metabolism between obese and lean individuáis may manifest themselves in functional adipose tissue abnormalities that lead to metabolic disorders in obesity. We studied lipogenesis and lipolysis of omental adipocytes from obese (OB) and non-obese (NOB) humans. The specific activity of the lipogenic marker enzyme G3PDH was 50 percent lower in total adipocytes of OB compared to that of NOB subjects. Omental adipocytes from OB subjects also had lower basal lipolytic levéis, and a lower lipolytic response to p-adrenergic stimulus. Cholesterol depletion of adipocyte plasma membrane using methyl β-cyclodextrin caused a lipolytic effect on adipocytes of both groups together, but when obese and lean subjects were analyzed separately, the response was significant only in the obese. We present evidence of a different lipogenic and lipolytic profile in obese individuáis' omental adipocytes, and propose a relevant role of plasma membrane cholesterol, where the impact of its removal in OB and NOB adipocyte lipolysis differs.


Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Adipocytes/cytology , Lipogenesis/physiology , Lipolysis/physiology , Obesity/physiopathology , Omentum/cytology , Adipocytes/physiology , Cholesterol/metabolism , Cholesterol/physiology , Membrane Lipids/physiology , Obesity/metabolism
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