Huazhi Rougan Granules attenuates steatosis in cell model of nonalcoholic fatty liver disease by inducing autophagy / 中国中药杂志

To investigate the effect of Huazhi Rougan Granules(HZRG) on autophagy in a steatotic hepatocyte model of free fatty acid(FFA)-induced nonalcoholic fatty liver disease(NAFLD) and explore the possible mechanism. FFA solution prepared by mixing palmitic acid(PA) and oleic acid(OA) at the ratio of 1∶2 was used to induce hepatic steatosis in L02 cells after 24 h treatment, and an in vitro NAFLD cell model was established. After termination of incubation, cell counting kit-8(CCK-8) assay was performed to detect the cell viability; Oil red O staining was employed to detect the intracellular lipid accumulation; enzyme-linked immunosorbnent assay(ELISA) was performed to measure the level of triglyceride(TG); to monitor autophagy in L02 cells, transmission electron microscopy(TEM) was used to observe the autophagosomes; LysoBrite Red was used to detect the pH change in lysosome; transfection with mRFP-GFP-LC3 adenovirus was conducted to observe the autophagic flux; Western blot was performed to determine the expression of autophagy marker LC3B-Ⅰ/LC3B-Ⅱ, autophagy substrate p62 and silent information regulator 1(SIRT1)/adenosine 5'-monophosphate(AMP)-activated protein kinase(AMPK) signaling pathway. NAFLD cell model was successfully induced by FFA at 0.2 mmol·L~(-1) PA and 0.4 mmol·L~(-1) OA. HZRG reduced the TG level(P<0.05, P<0.01) and the lipid accumulation of FFA-induced L02 cells, while elevated the number of autophagosomes and autophagolysosomes to generate autophagic flux. It also affected the functions of lysosomes by regulating their pH. Additionally, HZRG up-regulated the expression of LC3B-Ⅱ/LC3B-Ⅰ, SIRT1, p-AMPK and phospho-protein kinase A(p-PKA)(P<0.05, P<0.01), while down-regulated the expression of p62(P<0.01). Furthermore, 3-methyladenine(3-MA) or chloroquine(CQ) treatment obviously inhibited the above effects of HZRG. HZRG prevented FFA-induced steatosis in L02 cells, and its mechanism might be related to promoting autophagy and regulating SIRT1/AMPK signaling pathway.

Free fatty acids reduce metabolic stress and favor a stable production of heterologous proteins in Pichia pastoris

ABSTRACT The growth of yeasts in culture media can be affected by many factors. For example, methanol can be metabolized by other pathways to produce ethanol, which acts as an inhibitor of the heterologous protein production pathway; oxygen concentration can generate aerobic or anaerobic environments and affects the fermentation rate; and temperature affects the central carbon metabolism and stress response protein folding. The main goal of this study was determine the implication of free fatty acids on the production of heterologous proteins in different culture conditions in cultures of Pichia pastoris. We evaluated cell viability using propidium iodide by flow cytometry and thiobarbituric acid reactive substances to measure cell membrane damage. The results indicate that the use of low temperatures and low methanol concentrations favors the decrease in lipid peroxidation in the transition phase from glycerol to methanol. In addition, a temperature of 14 ºC + 1%M provided the most stable viability. By contrast, the temperature of 18 ºC + 1.5%M favored the production of a higher antibody fragment concentration. In summary, these results demonstrate that the decrease in lipid peroxidation is related to an increased production of free fatty acids.

Gut Microbial Influence and Probiotics on Colorectal Cancer / 대한소화기학회지

The human intestinal microbiota is a community of 10(13)-10(14) microorganisms that harbor in the intestine and normally participate in a symbiotic relationship with human. Technical and conceptual advances have enabled rapid progress in characterizing the taxonomic composition, metabolic capacity and immunomodulatory activity of the human intestinal microbiota. Their collective genome, defined as microbiome, is estimated to contain > or =150 times as many genes as 2.85 billion base pair human genome. The intestinal microbiota and its microbiome form a diverse and complex ecological community that profoundly impact intestinal homeostasis and disease states. It is becoming increasingly evident that the large and complex bacterial population of the large intestine plays an important role in colorectal carcinogenesis. Numerous studies show that gut immunity and inflammation have impact on the development of colorectal cancer. Additionally, bacteria have been linked to colorectal cancer by the production of toxic and genotoxic bacterial metabolite. In this review, we discuss the multifactorial role of intestinal microbiota in colorectal cancer and role for probiotics in the prevention of colorectal cancer.

Clinical profile of adrenoleukodysrophy.

X-linked Adrenoleukodystrophy (ALD) is the most common of the peroxisomal disorder and is associated with functional defect of the very long chain fatty acid (VLCFA) oxidation leading to the accumulation of VLCFA in the white matter and adrenal cortex. Retrospective evaluation of medical records of ALD patients were carried out. In all the 5 patients the duration of the symptoms varied from 1-7 years. Most of them presented with Addisonian crisis (4/5) and hyperpigmentation (5/5), white half of them (3/5) had neurological symptoms. All patients had biochemical evidence of the adrenal insufficiency. All siblings of patients should be screened for the possibility of ALD with VLCFA.

Molecular mechanism of insulin resistance.

Free fatty acids are known to play a key role in promoting loss of insulin sensitivity,thereby causing insulin resistance and type 2 diabetes.However,the underlying mechanism involved is still unclear.In searching for the cause of the mechanism,it has been found that palmitate inhibits insulin receptor (IR)gene expression,leading to a reduced amount of IR protein in insulin target cells. PDK1-independent phosphorylation of PKC(eta) causes this reduction in insulin receptor gene expression.One of the pathways through which fatty acid can induce insulin resistance in insulin target cells is suggested by these studies.We provide an overview of this important area,emphasizing the current status.

Epidemiología clínica del estado prediabético / Clinical epidemiology of the prediabetic state

La diabetes tipo 2 (DMT2) es un padecimiento muy común del metabolismo de los carbohidratos y los lípidos. La obesidad es un factor de riesgo significativo para el desarrollo de la diabetes tipo 2. La DMT2 es un factor de riesgo ampliamente reconocido para el desarrollo de la enfermedad arterial coronaria (EAC). Los pacientes con DMT2 tienen un incremento 2 a 4 veces mayor de cursar con EAC y un incremento 4 veces superior en el índice de mortalidad secundario a EAC. El estudio UKPDS y otros más han demostrado una mejoría significativa en el inicio y curso de las complicaciones microvasculares de la diabetes con un control glucémico apropiado. Sin embargo, esta aseveración no parece ser cierta con respecto a las complicaciones macrovasculares, ya que para ellas no existe una evidencia certera de una mejoría con un control apropiado en los niveles de glucosa en ayunas. Por lo anterior, es un hecho que un control intensivo del paciente diabético conlleva a un efecto mucho más modesto en reducir las complicaciones macrovasculares que las microvasculares. Esta aparente falta de benéficos en el paciente diabético al que se le ha mejorado su control glucémico ha dado lugar a intentar diferentes enfoques. Entre ellos se encuentra el que pretende prestar mayor atención al paciente obeso, prediabético caracterizado por la presencia de resistencia a la insulina. Esta última se asocia a un número significativo de anormalidades protrombóticas y proinflamatorias, que incluyen un exceso de grasa corporal, que podrían jugar un papel causal en el desarrollo de la enfermedad cardiovascular. Por lo dicho, el tratamiento de la obesidad y la prevención del desarrollo de la diabetes tipo 2 parecen ser estrategias prioritarias para prevenir o retardar la aparición de las enfermedades cardiovasculares.

Effect of High Fat Diet on Insulin Resistance: Dietary Fat Versus Visceral Fat Mass

The purpose of the present study was to determine whether chronic high-fat diet (HF) induces insulin resistance independently of obesity. We randomly divided 40 rats into two groups and fed them either with a HF or with a high-carbohydrate diet (HC) for 8 weeks. Whole body glucose disappearance rate (Rd) was measured using a euglycemic hyperinsulinemic clamp. Firstly, we defined whether insulin resistance by HF was associated with obesity. Plasma glucose and triglyceride concentrations were significantly increased in HF. Rd was decreased (10.6+/-0.2 vs. 9.1+/-0.2 mg/kg/min in HC and HF, respectively) and the hepatic glucose output rate (HGO) was increased in HF (2.2+/-0.3 vs. 4.5+/-0.2 mg/kg/min in HC and HF, respectively). Rd was significantly correlated with %VF (p<0.01). These results implicate that visceral obesity is associated with insulin resistance induced by HF. In addition, to define whether dietary fat induces insulin resistance regardless of visceral obesity, we compared Rd and HGO between groups 1) after matching %VF in both groups and 2) using an ANCOVA to adjust for %VF. After matching %VF, Rd in HF was significantly decreased by 14% (p<0.001) and HGO was significantly increased by 110% (p<0.001). Furthermore, statistical analyses using an ANCOVA also showed Rd for HF was significantly decreased even after adjusting %VF. In conclusion, we suggest that dietary fat per se could induce insulin resistance in rats fed with chronic HF independently of obesity.

Effect of ethanol/arrack on the lipid metabolism of mammary gland during pregnancy and lactation in rats.

Female rats were exposed to arrack (12.0 ml/kg body weight/day) and ethanol (4.0 g/kg body weight/day) before conception and throughout gestation and lactation. On 19th day of gestation and 21st day of lactation there was increase in the cholesterol phospholipids, triglycerides and free fatty acids in the mammary gland of rats administered arrack/ethanol in comparison with the controls. The lipoprotein lipase activity showed significant increase in the treated groups, in which the activity decreased on 21st day in comparison with 19th day. The absolute and relative weight of mammary gland also showed a significant decrease in ethanol/arrack treated group. The biochemical alterations produced in the mammary gland by arrack and its equivalent alcohol were different showing that non-alcoholic portion of arrack interferes with the toxicity induced by alcohol. Arrack was found to be a potent hyperlipidemic agent than ethanol.

Coriandrum sativum changes the levels of lipid peroxides and activity of antioxidant enzymes in experimental animals.

The antiperoxidative effect of coriander seeds (Coriandrum sativum) was studied in rats administered high fat diet. Significant decrease in the levels of lipid peroxides, free fatty acids and glutathione was observed when compared to control group whereas the activity of antioxidant enzymes showed increase.

Decreased phospholipase A2 activity in butyrate differentiated HT29 cells.

Our earlier work has shown that in butyrate differentiated colonic HT29 cells, there is an alteration in phospholipid composition as compared to control. To know more about these changes, butyrate treated and control cell homogenates were incubated in presence of calcium and phospholipids were analyzed. It was observed that incubation with calcium was associated with increase in lysophosphatidylcholine (lysoPC) and free fatty acids and the increase was much higher in control as compared to butyrate treated cells. There was no alteration in lysoPC content. These products are formed by the action of phospholipase A2 (PLA2) which is activated by calcium and suggests that butyrate-induced differentiation is associated with decrease in PLA2 activity.

Danazol-induced biochemical changes in the rat ovary.

Effect of varying doses of danazol, a synthetic steroid derivative of 17 alpha-ethinyl testosterone has been observed on the biochemistry of the rat ovary. Biochemically total proteins decreased and total lipids increased with the danazol treatment. Triglycerides, the stored form of lipids formed the major components of lipids in the treated ovaries. The amount of phospholipids, glycolipids, cholesterol and free fatty acids decreased in the ovaries with increased danazol treatment. The functional significance of these changes have been discussed with ovarian physiology especially in relation to follicular growth and atresia.

Free fatty acids (FFA) and type II diabetes

Clinical and experimental data indicate that there is an increase in plasma FFA levels in type II diabetes. Despite the enhanced lipolysis all evidences suggested no rate of entry of FFA into plasma, implying a compensatory increase in re-esterification of FFA liberated from adipose tissue, a reduced clearance being the most likely explanation for the increase in plasma FFA in NIDDM. The increased FFA availability can impair insulin-mediated glucose disposal and increase hepatic glucose production probably mediated by gluconeogenesis. Finally, while the incresed FFA levels induce insulin resistance in peripheral tissues, particularly muscle, there is a stimulation of insulin secretion in order to match the insulin resistance. However, the chronic FFA overload impairs the B-cells and the hyperinsulinemia no longer matches the increase in insulin resistance and NIDDM begins.

Lipid effects on nicotinic acetylcholine receptor gating and kinetics: a structural-functional correlation

The nicotinic acetylcholine receptor (AChR) is still the paradigm of rapid ligand-gated ion channels. Since the early finding of a motionally restricted shell of lipids ( annulus ) in the immediate perimeter of the membrane-bound AChR, experimental evidence has supported the notion that the interface between the protein moiety and the adjacent lipid molecules is the site of action of a variety of pharmacologically relevant substances, including non-competitive inhibitors of the cholinergic system like some local anesthetics, short-chain alcohols, and steroids. Patch-clamp data on cells expressing the AChR protein add another dimension to this knowledge, enabling correlations to be established between the chemical composition of lipid-modified cells and the functional properties (ligand binding, channel gating) of the receptor protein in situ

Effect of aspirin on isoproterenol induced changes in lipid metabolism in rats.

The effect of aspirin on isoproterenol induced changes in lipid metabolism in rats was studied. Aspirin (1.2 mg/100 g/day) was administered orally for a period of 60 days along with/without isoproterenol (20 mg/100g sc twice at a time interval of 24 h for 2 days). Isoproterenol treated rats showed an increase in the levels of heart cholesterol, triglycerides and free fatty acids. The activity of cholesterol ester synthetase CES was increased significantly with concomitant increase in heart lipid peroxide levels in isoproterenol treatment. Aspirin treatment could restore the enzyme activity to near normal and also reduce the level of lipid peroxides. The lipid changes were minimum in rats treated with aspirin and isoproterenol.

Efectos endócrinos de la hipnosis con tiopental sódico en los perros / Endocrine effects of thiopental anaesthesia in dogs

se estudiaron los efectos endócrinos de la hipnosis con tiopental sódico (TPS) en perros con respiración espontánea (RE) y controlada mecánicamente (RC) durante tres horas. Se emplearon 18 perros divididos en tres grupos: 1-controles no anestesiados (CNA). 2-anestesiados con TPS y RE y 3-anestesiados con TPC y RC. Se controló la: tensión arterial media, frecuencia cardíaca, frecuencia respiratoria y temperatura rectal. Se efectuaron extracciones de sangre antes de la inducción y luego periódicamente para controlar la glucemia y los niveles de: ácidos grasos no esterificados ACTH, cortisol, catecolaminas, insulina, T3, T4 junto con las concentraciones plasmáticas de TPS. También se monitoreó el pH, gases en sangre, exceso de base y bicarbonato standard. La técnica anestésica descrita con RE y RC no modificó los niveles séricos y plasmáticos de insulina, T3 y catecolaminas, pero provocó una disminución importante en los ácidos grasos no esterificados séricos y en las concentraciones de T4, sin embargo, no logró suprimir totalmente la respuesta tardía del eje hipófiso-suprarrenal al stress anestésico.

Peripheral glucose and potassium transport after glucose ingestion in normal subjects

This study was designed to qualify forearm potassium uptake or release in the postabsorptive state and after an oral glucose challenge, and if there is association of potassium uptake with glucose and free fatty acids(FFA) metabolism in peripheral tissues of normal subjects. Nine helathy volunteers (4M and 5F) were stidied after an overnight fast (12-14 h) and during 3 hours after ingestion of 75 g of glucose. The forearm glucose uptake rat reached the maximum at 60 minutes (0.985 mg/100 ml forearm.min), declining to the basal levels at the end of the 3 h of study. Although heterogeneously in the postabsorptive state, potassium moves out of the forearm muscle into the venous blood in normal individuals. After glucose ingestion, the potassium release diminished and there was also potassium uptake during the last hour of the study. The arterial FFA levels decreased significantly after the glucose liad. In conclusion, the results of this study showed that in peripheral tissues: 1) during the first 2 h after the ingestion of 75 g of glucose ther was a reduction in potassium release; 2) potassium uptake only occurred in the last hour when insulin levels were slightly higher, plasma glucose were similar and FFA were lower than in the postabsorptive period; 3) there was a clear dissociation between potassium and glucose uptake

Metabolic adjustment induced by exposure to elevated ambient temperature is impaired in rats bearingt lesions in the preoptic area

Metabolic adjustment was studied in rats with bilateral electrolytic lesions in the preoptic area which were exposed to heat (34-C) anda to normal temperature (25-C) for 60 days. The metabolic rate measured at neutral temperature (29-C) in preoptic-lesioned rats living at 25- or 34-C was reduced to 80% the rate of sham-operated controls. The increase in metabolic rate induced by heat (30 min at 34- or 37-C) in preoptic-lesioned rats, living at 25-C, was markedly reduced to 10% the rate of sham-operated controls living at 25-C. In preoptic-lesioned rats, the increased metabolic rate induced by heat exposure (34- or 37-C) was impaired, regardless of the living ambient temperature (25- or 34-C) of the animal. These data demonstrate the participation of the preoptic area in heat-induced invreased oxygen consumption and provide additional evidence for its role in the integration of thermal imputs with energy-linked metabolic processes

Effect of lipid infusion on substrate uptake in soleus muscle of the cat

Se estudió la capacitación de glucosa, ácidos grasos libres (AGL) y ácido láctico (AL) en el músculo soleo de gatos anestesiados y heparinizados. Los animales se estudiaron en tres grupos: 1) gatos enjaulados, 2) gatos "libres", 3) gatos entrenados. Muestras de sangre arterial y del efluente venoso del músculo se tomaron a) en reposo b) después de la contracción muscular c) después de una infusión con Intralipid y d) después de repetir la contracción luego de haber efectuado la infusión. En a) se encontró una correlación directa entre los niveles arteriales de AGL y su captación, en todos los grupos, y una relación directa entre la captación de glucosa y producción de AL en los gatos entrenados. En b) y d) los niveles de AGL estuvieron en relación directa con la producción de AL. En conclusión el soleo del gato en las condiciones estudiadas utiliza preferencialmente los AGL; en la contracción usa el glucógeno con aumento no significativo de la producción de AL. La correlación entre los niveles venosos de AGL y el AL sugiere que los aGL inhiben la entrada de piruvato al ciclo de loa ácidos tricarboxílicos

Controle pancreático do metabolismo de carboidratos e de lipídios no pombo / Pancreatic control of carbohydrate and lipid metabolism in the pigeon

O presente trabalho relata o papel do pâncreas sobre os níveis de ácidos graxos livres (AGL) e de triglicerídios plasmáticos (TG), e sobre a glicemia de pombos. Discute-se também a resposta pós-insulínica dos níveis de AGL nestas aves pancreatectomizadas ou pancreatecto-enterectomizadas. Verifica-se que em pombos a administraçäo de insulina endovenosa aumenta os níveis de AGL e diminui os níveis glicêmicos, e o glucagon eleva os níveis de AGL e glicêmicos. Nas aves pancreatectomizadas ocorre hiperglicemia transitória, queda do TG plasmáticos, näo se alterando os níveis de AGL. A administraçäo de insulina nas aves pancreatectomizadas ou pancreatecto-enterectomizadas eleva as taxas de AGL e diminui a glicemia. A resposta pós-insulina dos níveis de AGL deve estar na dependência de outros fatores que näo o glucagon entérico ou pancreático, possivelmente das adrenais, bem como do sistema nervoso simpático