Ma.Concepción Gutiérrez-Ruiz

Health-related quality of life and depression in patients with chronic hepatitis C

BACKGROUND Hepatitis C is a major cause of liver disease worldwide. It has been associated with decreased health-related quality of life (HRQL) and psychiatric symptoms. Our aim was to assess HRQL, depression, and illness understanding in patients with chronic hepatitis C without previous interferon therapy. METHODS Consecutive patients attending a referral center were enrolled. HRQL was measured using SF-36 questionnaire, depression with Zung self-rating depression scale, and illness understanding with self-applied knowledge test. RESULTS Of 157 patients enrolled, 112 were female (71%) and 45 male (29%). Ninety-seven patients (61.8%) had cirrhosis. HRQL was significantly decreased in chronic hepatitis C patients compared to historical normal controls in all eight domains of the SF-36 (p < 0.001). In hepatitis C cirrhotic patients, HRQL was significantly lower among Child-Pugh class B and C subjects in domains reflecting physical health (p <0.05). Ninety-two patients (58.6%) had depression that resulted in lower HRQL when compared to nondepressed patients (p <0.05). One hundred fourteen patients (72.6%) had poor illness understanding of hepatitis C. These subjects had significantly lower HRQL scores in six of eight SF-36 domains when compared to patients with better understanding of the disease (p <0.05). CONCLUSIONS Chronic hepatitis C patients attending a tertiary-referral center had significant decrease in HRQL associated with depression (58.6%) and poor illness understanding (72.6%). Educational programs and their impact on HRQL need to be addressed in detail, particularly for the pre-treatment scenario.

Cytokines, growth factors, and oxidative stress in HepG2 cells treated with ethanol, acetaldehyde, and LPS

Inflammatory mediators, including cytokines, growth factors, and reactive oxygen species, are associated with the pathology of chronic liver disease. In the liver, cytokine and growth factor secretion are usually associated with nonparenchymal cells, particularly Kupffer cells. In the present studies, the effect of 24 and 72 h administration of ethanol (50 mM). acetaldehyde (175 microM), and LPS (1 microg/ml) were studied on the expression and secretion of TNF-alpha, IL-1beta, IL-6, and TGF-beta3, lipid peroxidation damage and glutathion content in HepG2 cell cultures. A 24 h exposure to ethanol induced the expression of TNF-alpha and TGF-beta1, and the secretion of IL-1beta and TGF-beta1. With the same period of treatment, acetaldehyde markedly increased TNF-alpha expression, and stimulated IL-1beta secretion, while LPS exposure induced the expression of TNF-alpha, IL-6, and TGF-beta1, and the secretion of IL-1beta, IL-6, and TGF-beta1. A reduced in TNF-alpha response and TGF-beta1 expression were observed after 72 h exposure to ethanol. A 72 h acetaldehyde exposure decreased markedly TNF-alpha expression and stimulated a previously absent TGF-beta1 response. With the same time of exposure, LPS reduced slightly TGF-beta1 expression, and decreased its secretion. IL-1beta and IL-6 were not detected under 72 h exposure conditions. Lipid peroxidation damage was increased in all treatments, but higher values were found in 72 h treatments. Glutathion content diminished in all treatments. These findings suggest that HepG2 cells, independent of other cells such as Kupffer or macrophages, participate in a differential cytokine, growth factor and oxidative stress response, which differs according to the toxic agent and the time of exposure.

Uptake, cellular distribution and DNA damage produced by mercuric chloride in a human fetal hepatic cell line

A human hepatic cell line (WRL-68 cells) was employed to investigate the uptake of the toxic heavy metal mercury. Hg accumulation in WRL-68 cells is a time and concentration dependent process. A rapid initial phase of uptake was followed by a second slower phase. The transport does not require energy and at low HgCl2 concentrations (<50 microM) Hg transport occurs by temperature-insensitive processes. Subcellular distribution of Hg was: 48% in mitochondria, 38% in nucleus and only 8% in cytosolic fraction and 7% in microsomes. Little is known at the molecular level concerning the genotoxic effects following the acute exposure of eucaryotic cells to low concentrations of Hg. Our results showed that Hg induced DNA single-strand breaks or alkali labile sites using the single-cell gel electrophoresis assay (Comet assay). The percentage of damaged nucleus and the average length of DNA migration increased as metal concentration and time exposure increased. Lipid peroxidation, determined as malondialdehyde production in the presence of thiobarbituric acid, followed the same tendency, increased as HgCl2 concentration and time of exposure increased. DNA damage recovery took 8 h after partial metal removed with PBS-EGTA.

Lower Serum IL-10 Is an Independent Predictor of IBS Among Volunteers in Mexico

OBJECTIVES:Studies suggest that altered immune activation, manifested by an imbalance in anti- and pro-inflammatory cytokine levels, exists in a subgroup of irritable bowel syndrome (IBS) patients. However, similar studies have not been conducted in Latin populations. The objective of this study was to measure serum levels of interleukin (IL)-10 and tumor necrosis factor (TNF)-α in subjects fulfilling symptom criteria for IBS and controls.METHODS:Volunteers (n=178) from a university population in Mexico City, participated in the study. Of the sample, 34.8% met Rome II criteria for IBS and 65.2% were designated as controls. Serum cytokines were measured by enzyme-linked immunoabsorbent assay. Analysis of covariance models were used to test main effects between gender, IBS symptoms, and bowel habit subtype to explain the cytokine serum levels. Statistical models were tested using body mass index as a covariate.RESULTS:IL-10 levels were significantly lower in IBS vs. controls (mean (95% confidence interval): 15.6 (14.8, 16.3) vs. 18.6 (17.9, 19.4) pg/ml, P<0.001), while TNF-α levels were higher in IBS (20.9 (19.1, 23.0) vs. 17.9 (16.7, 19.3) pg/ml, P=0.010). IBS and female gender were independent predictors for IL-10 (P<0.05). In contrast, female gender was an independent predictor for TNF-α. In addition, women with IBS-D had the lowest IL-10 (P<0.001) and highest TNF-α (P=0.021) vs. other subtypes.CONCLUSIONS:The lower serum IL-10 in our subjects fulfilling IBS Rome II symptom criteria suggests an altered immune regulation. Further studies are needed to elucidate if a lower serum IL-10 may be useful as a biomarker for IBS in the Mexican population, especially for women with IBS-D.

Liver fibrosis: searching for cell model answers

Hepatic stellate cells (HSC) are the principal fibrogenic cell type in the liver. Progress in understanding the cellular and molecular basis for the development and progression of liver fibrosis could be possible by the development of methods to isolate HSC from rodents and human liver. Growth of stellate cells on plastic led to a phenotypic response known as activation, which paralleled closely the response of these cells to injury in vivo. Actually, much of the current knowledge of stellate cell behaviour has been gained through primary culture studies, particularly from rats. Also, different laboratories that have established hepatic stellate cell lines from rats and humans have provided a stable and unlimited source of cells that express specific functions, making them suitable for culture‐based studies of hepatic fibrosis. From these in vitro models grew a large body of information characterizing stellate cell activation, cytokine signalling, intracellular pathways regulating liver fibrogenesis, production of extracellular matrix proteins and development of antifibrotic drugs.

Cadmium induces α1collagen (I) and metallothionein II gene and alters the antioxidant system in rat hepatic stellate cells

Abstract The mechanism of cadmium-mediated hepatotoxicity has been the subject of numerous investigations, principally in hepatocytes. Although, some uncertainties persist, sufficient evidence has emerged to provide a reasonable account of the toxic process in parenchymal cells. However, there is no information about the effect of cadmium in other hepatic cell types, such as stellate cells (fat storing cells, Ito cells, perisinusoidal cells, parasinusoidal cells, lipocytes). Hepatic stellate cells (HSC) express a quiescent phenotype in a healthy liver and acquire an activated phenotype in liver injury. These cells play an important role in the fibrogenic process. The objective of this study was to investigate the effect of a 24 h treatment of low Cd concentrations in glutathione content, lipid peroxidation damage, cytosolic free Ca, antioxidant enzyme activities: glutathione peroxidase, glutathione reductase, superoxide dismutase and catalase along with the capacity of this heavy metal to induce metallothionein II and α 1 collagen (I) in an hepatic stellate cell line (CFSC-2G). Cd-treated cells increased lipid peroxidation and the content of cytosolic free calcium, decreased glutathione content and superoxide dismutase, glutathione peroxidase and catalase activity. Cd was able to induce the expression of the metallothionein II and α 1 collagen (I) gene, that was not described in this cell type. Cadmium may act as a pro-fibrogenic agent in the liver probably by inducing oxidative damage by enhancing lipid peroxidation and altering the antioxidant system of the cells. Although, the exact role metallothionein induction plays in this process is unknown, it probably, provides a cytosolic pool of potential binding sites to sequester ionic Cd, thereby decreasing its toxicity.

Interleukin 8 response and oxidative stress in HepG2 cells treated with ethanol, acetaldehyde or lipopolysaccharide

The aim of this work was to study the induction and secretion of interleukin 8 (IL-8) and some oxidative stress parameters after ethanol (EtOH), acetaldehyde (Ac) or lipopolysaccharide (LPS) treatment on HepG2 cells. Cells were treated with 50 mM EtOH, 175 &mgr;M Ac or 1 &mgr;g/ml of LPS. IL-8 induction and secretion were determined in the presence of the toxics, and the effect of antioxidants N-acetyl-L-cysteine and 1,1,3,3-tetramethyl-2-thiourea was evaluated. Further, the effect of adding polyclonal anti-human tumor necrosis factor alpha (TNF-alpha) and H(2)O(2) was studied, and catalase, superoxide dismutase and glutathione peroxidase activities were determined. Lipid peroxidation increased significantly only in Ac-treated cells. All toxics failed to decrease significantly the intracellular levels of reduced GSH. Catalase activity was diminished in all treatments, while other enzyme activities did not present changes. No change in peroxide production was found with any treatment. IL-8 secretion increased in Ac (41%) and in LPS (38%)-treated cells. Antioxidant and anti-TNF-alpha treatments decreased IL-8 secretion. H(2)O(2) (0.25 mM)-treated cells increased IL-8 secretion. IL-8 reverse transcriptase-polymerase chain reaction results correlated with secretion values. Our results show that Ac and LPS treatment produced an increased IL-8 induction and secretion. Oxidative stress and TNF-alpha are mediators in IL-8 response. This observation suggests that in the in vivo liver, the mechanism of ethanol-induced IL-8 production requires ethanol metabolism, and hepatocytes do not require the interaction among different populations of liver cells to respond.

Effect of endotoxin pretreatment on hepatic stellate cell response to ethanol and acetaldehyde

The role of endotoxin in alcohol‐induced liver damage is well recognized. How pre‐exposure to endotoxin might affect alcohol injury is not known. We herein studied the effect of endotoxin pretreatment on hepatic stellate cell (HSC) response to ethanol and acetaldehyde.

DNA damage produced by cadmium in a human fetal hepatic cell line

Cadmium (Cd) is one of the most important heavy metal environmental toxicants. It alters a wide variety of cellular and biochemical processes. The objective of this work was to study DNA damage and recovery after acute and chronic CdCl2 treatment in a human fetal hepatic cell line (WRL-68 cells). Using the alkaline microgel electrophoresis assay that detects DNA single-strand breaks and/or alkali-labile sites in individual cells, we evaluated for levels of DNA damage. The mean migration length in control cells was 35.37+/-1. 43 microm (8% damaged cells), whereas the mean migration in cells treated with 0.005 microM CdCl2 for 3 h (acute low dose) was 65. 87+/-2.07 microm (88% damaged cells). Treatment with 0.01 microM CdCl2 for the same time (acute high dose) increased the mean migration length to 125.79+/-2.91 microm (92% damaged cells). However, a 0.005 microM CdCl2 treatment for 7 days (chronic treatment) only increased 65% DNA migration to 58.38+/-2.59 microm (88% damaged nucleus). Lipoperoxidative damage expressed as malondialdehyde (MDA) production per milligram of protein was 15. 7+/-2.6 for control cells, whereas in Cd-treated cells the values were 20.2+/-2.4 (acute low dose), 22.9+/-2.2 (acute high dose), and 22.6+/-2.1 (chronic treatment). To study the repair of DNA damage, cells were washed with 0.01 microM meso-2,3-dimercaptosuccinic acid (DMSA), and fresh Dulbeccos modified essential medium (DMEM) added. The percentage of damaged cells diminished after 90 min, with DNA migration returning to control values by 120 min. Cd treatment produced DNA single-strand breaks and the damage was greater in acute high dose treated cells. Lipid peroxidation values did not correlate with DNA single-strand breaks.

Emerging concepts in inflammation and fibrosis

Hepatic and pancreatic response to several insults commonly includes similar pathways of inflammation, fibrogenesis/regeneration, which may occur simultaneously and without appropriate coordination, resulting in chronic inflammation, scarring, and organ dysfunction. This review highlights the opinion of experts gathered for the Mexican Digestive Disease Week (2001) to analyze these molecular events with emphasis on identifying possible therapeutic opportunities. Inflammatory response encompasses leukocyte infiltration, favored by adhesion molecules of the selectin family, chemokines, integrins, and activated stellate cells (SC). Quiescent SC undergo activation mediated by mechanical stress and expression of cytokines, oxidative stress products, and growth factors and play a significant role in fibrosis and in reparation toward synthesis of extracellular matrix components. Also, hepatocytes and acinar cells contribute to the inflammatory and fibrotic response. Molecules that down-regulate this response are overexpressed. Therapeutic strategies with targeting to such mechanisms underlying chronic hepatic and pancreatic injury are an emerging reality.