Differences in Hepatic Expression of Iron, Inflammation and Stress-Related Genes in Patients with Nonalcoholic Steatohepatitis

Abstract: Nonalcoholic fatty liver disease (NAFLD) is the most common cause of chronic liver disease worldwide. We have previously shown that hepatic reticuloendothelial system (RES) iron deposition is associated with an advanced degree of nonalcoholic steatohepatitis (NASH) in humans. In this study, we aimed to determine differentially expressed genes related to iron overload, inflammation and oxidative stress pathways, with the goal of identifying factors associated with NASH progression. Seventy five patients with NAFLD were evaluated for their biochemical parameters and their liver tissue analyzed for NASH histological characteristics. Gene expression analysis of pathways related to iron homeostasis, inflammation and oxidative stress was performed using real-time PCR. Gene expression was compared between subjects based on disease status and presence of hepatic iron staining. We observed increased gene expression of hepcidin (HAMP) (2.3 fold, p = 0.027), transmembrane serine proteinase 6 (TMPRSS6) (8.4 fold, p = 0.003), signal transducer and activator of transcription 3 (STAT3) (5.5 fold, p = 0.004), proinflammatory cytokines; IL-1β (2.7 fold, p = 0.046) and TNF-α (3.8 fold, p = 0.001) in patients with NASH. TMPRSS6, a negative regulator of HAMP, is overexpressed in patients with NASH and HIF1α (hypoxia inducible factor-1) is downregulated. NAFLD patients with hepatic iron deposition exhibited higher hepcidin expression (3.1 fold, p = 0.04) but lower expression of cytokines. In conclusion, we observed elevated hepatic HAMP expression in patients with NASH and in NAFLD patients who had hepatic iron deposition, while proinflammatory cytokines displayed elevated expression only in patients with NASH, suggesting a regulatory role for hepcidin in NAFL to NASH transition and in mitigating inflammatory responses.

Noninvasive Assessment of Hepatic Fibrosis in Patients with Chronic Hepatitis B Viral Infection Using Magnetic Resonance Elastography

OBJECTIVE: To evaluate the diagnostic performance of magnetic resonance elastography (MRE) for staging hepatic fibrosis in patients with chronic hepatitis B virus (HBV) infection. MATERIALS AND METHODS: Patients with chronic HBV infection who were suspected of having focal or diffuse liver diseases (n = 195) and living donor candidates (n = 166) underwent MRE as part of the routine liver MRI examination. We measured liver stiffness (LS) values on quantitative shear stiffness maps. The technical success rate of MRE was then determined. Liver cell necroinflammatory activity and fibrosis were assessed using histopathologic examinations as the reference. Areas under the receiver operating characteristic curve (Az) were calculated in order to predict the liver fibrosis stage. RESULTS: The technical success rate of MRE was 92.5% (334/361). The causes of technical failure were poor wave propagation (n = 12), severe respiratory motion (n = 3), or the presence of iron deposits in the liver (n = 12). The mean LS values, as measured by MRE, increased significantly along with an increase in the fibrosis stage (r = 0.901, p or = F1, > or = F2, > or = F3, and F4 were 2.45 kPa, 2.69 kPa, 3.0 kPa, and 3.94 kPa, respectively, and with Az values of 0.987-0.988. CONCLUSION: MRE has a high technical success rate and excellent diagnostic accuracy for staging hepatic fibrosis in patients with chronic HBV infection.

Sobrecarga de ferro em adolescente com xerocitose: a importância da ressonância nuclear magnética / Iron overload in a teenager with xerocytosis: the importance of nuclear magnetic resonance imaging

Relatar um caso de sobrecarga de ferro secundária à xerocitose, uma doença rara, em uma adolescente, diagnosticada por meio de ressonância magnética em T2*. Relatamos o caso de uma paciente sintomática com xerocitose, nível de ferritina de 350ng/mL e sobrecarga de ferro cardíaca significativa. Ela foi diagnosticada por ressonância magnética em T2* e recebeu terapia de quelação. Análise por ectacitometria confirmou o diagnóstico de xerocitose hereditária. Na sequência, a ressonância magnética em T2* demonstrou resolução completa da sobrecarga de ferro em vários órgãos e novo ecocardiograma revelou resolução completa das alterações cardíacas anteriores. A paciente permanece em terapia de quelação. Xerocitose é uma desordem genética autossômica dominante rara, caracterizada por estomatocitose desidratada. O paciente pode apresentar fadiga intensa e sobrecarga de ferro. Sugerimos o uso regular de ressonância magnética em T2* para o diagnóstico e controle da resposta à quelação de ferro em xerocitose e acreditamos que o exame pode ser útil também em outras anemias hemolíticas que necessitam de transfusões.

To report a case of iron overload secondary to xerocytosis, a rare disease in a teenager, diagnosed, by T2* magnetic resonance imaging. We report the case of a symptomatic patient with xerocytosis, a ferritin level of 350ng/mL and a significant cardiac iron overload. She was diagnosed by T2* magnetic resonance imaging and received chelation therapy Ektacytometric analysis confirmed the diagnosis of hereditary xerocytosis. Subsequent T2* magnetic resonance imaging demonstrated complete resolution of the iron overload in various organs, as a new echocardiography revealed a complete resolution of previous cardiac alterations. The patient remains in chelation therapy. Xerocytosis is a rare autosomal dominant genetic disorder characterized by dehydrated stomatocytosis. The patient may present with intense fatigue and iron overload. We suggest the regular use of T2* magnetic resonance imaging for the diagnosis and control of the response to iron chelation in xerocytosis, and we believe it can be used also in other hemolytic anemia requiring transfusions.

Valor utilitario específico de la técnica modificada de Perls para el diagnóstico anatomopatológico / Specific utilitarian value of the modified technique of Perls for the pathological diagnosis

Se modificó la técnica de Perls para disponer de un medio que pudiera revelar la presencia del pigmento férrico y poder utilizarlo en el diagnóstico histopatológico; para ello se elevó a 58 grados centígrados la temperatura en los reactivos, lo cual no se hace en el procedimiento habitual, y el citado pigmento se tiñó de verde o azul, de manera que fue posible identificarlo en los núcleos grises de la base en un paciente con síndrome de Hallervordem - Spatz, que falleció en esta institución.

The technique of Perls was modified to have a mean that could reveal the presence of the ferric pigment and to be able to use it in the histopathological diagnosis; for this purpose the temperature was rised to 58º centigrades in the reagents, which is not done in the habitual procedure, and the mentioned pigment was stained of green or blue, so that it was possible to identify it in the gray nuclei of the base in a patient with syndrome of Hallervordem - Spatz, who died in this institution.

Assessmrent of iron overload in thalassaemic patients by magnetic resonance imaging: a pilot study.

The patients of thalassaemia major need repeated blood transfusion which leads to excess iron deposition in various organs like liver, heart, pituitary etc. This iron accumulation causes various complications and ultimately organs' failure. There is no non-invasive, standard and reliable method to know the status of iron overload in various organs of the body. This paper attempts to use magnetic resonance imaging to know the liver iron overload in 8 thalassaemic patients as a pilot study. Eight children suffering with thalassaemia and 3 controls who were the normal siblings of the patient group underwent magnetic resonance imaging of the abdomen using spin-echo T, weighted sequence. Blood serum ferritin levels in the patients' group were also determined on the same day of magnetic resonance imaging examination. It was observed that the ratio of magnetic resonance imaging signal intensity (in spin-echo T1 weighted image) in paraspinous muscle to liver was significantly different in normal control (0.65) compared to that in thalassaemia patients (2.1 to 11.4 depending upon extent of iron deposition). The magnetic resonance signal intensity ratio correlated with the blood serum ferritin level of patients (p = 0.01) which is generally taken as indirect measure of body iron burden. Spin-echo sequence is the simplest imaging sequence and it increases the chance of its routine use. The study concludes that magnetic resonance imaging has good potential to quantify the liver iron deposition non-invasively and may denote the efficacy of iron-chelation therapy which is used to reduce the body iron burden in these patients.

Hereditary hemochromatosis in a Brazilian university hospital in São Paulo State (1990-2000)

Hereditary hemochromatosis (HH) is the most common genetic disease among individuals of European descent. Two mutations (845G-->A, C282Y and 187C-->G, H63D) in the hemochromatosis gene (HFE gene) are associated with HH. About 85-90% of patients of northern European descent with HH are C282Y homozygous. The prevalence of HH in the Brazilian population, which has a very high level of racial admixture, is unknown. The aims of the present study were to identify individuals with diagnostic criteria for HH among patients with a body iron overload attended at the university hospital of the Faculty of Medicine of Ribeirao Preto from 1990 to 2000, and to evaluate the prevalence of HFE mutations. We screened first-degree relatives for HFE mutations. Four of 72 patients (three men and one woman, mean age 47 years) fulfilled the criteria for HH. HFE mutations were studied in three patients [two C282Y homozygotes (patients 1 and 2) and one H63D heterozygote]. Patient 1 had four children (all C282Y heterozygotes with no iron overload) and seven brothers and sisters: two sisters (66 and 76 years old) were C282Y homozygotes and both had an iron overload (a liver biopsy in one showed severe iron deposits), one sister (79 years old) was a compound heterozygote with no iron overload, one brother (78 years old) was a C282Y heterozygote with no iron overload, two individuals were H63D heterozygotes (one brother, 49 years old, obese, with a body iron overload and abnormal liver enzymes - a biopsy showed non-alcoholic steatohepatitis, and one 70-year-old sister with no iron overload). Patient 2 had two children (22 and 24 years old who were C282Y heterozygotes with no iron overload) but no brothers or sisters. These results showed that HH was uncommon among individuals attended at our hospital, although HFE mutations were found in all patients. Familial screening is valuable for the early diagnosis of individuals at risk since it allows treatment to be initiated before the onset of the clinical manifestations of organ damage associated with HH

T[2] relaxation time of the pituitary gland in patients with beta-thalassemia and its relation to iron overload and somatic and sexual development

Chelation therapy with deferoxamine mesylate [has revolutionized management of transfusion-dependent beta-thalassaemia, but monitoring of tissue iron deposition, particularly in the endocrine glands, is still largely empirical. Clinical, haematological, and endocrinal evaluation of 54 transfusion-dependent beta-thalassaemic patients and 25 age- and sex-matched controls was done, pituitary T2 relaxation time was studied in them by Magentic Resonance Imaging [MRI] to evaluate pituitary iron overload. Thalassaemic patients had significantly lower mean stature, growth velocity, and a more delayed pubertal stage. Sixty per cent of thalassaemic females had amenorrhoea, either 1 ry or 2ry. Serum insulin-like growth factor 1 [IGF-1] and lutenizing hormone [LH] were significantly lower in thalassaemic patients compared with controls. Serum growth hormone [and follicule stimulating hormone [GH] were also lower, but the difference was not statistically significant. Pituitary T2 relaxation rate was significantly higher in patients compared with controls. Serum ferritin in thalassaemic patients showed a statistically significant positive correlation with pituitary T2 relaxation time, and a statistically significant negative correlation with serum IGF-1. It was concluded that monthly follow up of haemoglobin level and serum ferritin are vital to guide a satisfactory transfusion/chelation regimen in thalassaemic patients. However, once a deviation arises in physical/sexual development, measurement of GH, IGFI, FSH, and LH is warranted. Patients with clinical and/or laboratory evidence of pituitary dysfunction will benefit from an MRI study to assess pituitary iron deposition and provide a better guide for the intensity of chelation therapy