Anti-Inflammatory Effects of a Pomegranate Leaf Extract in LPS-Induced Peritonitis.

Folk medicine suggests that pomegranate (peels, seeds and leaves) has anti-inflammatory properties; however, the precise mechanisms by which this plant affects the inflammatory process remain unclear. Herein, we analyzed the anti-inflammatory properties of a hydroalcoholic extract prepared from pomegranate leaves using a rat model of lipopolysaccharide-induced acute peritonitis. Male Wistar rats were treated with either the hydroalcoholic extract, sodium diclofenac, or saline, and 1 h later received an intraperitoneal injection of lipopolysaccharides. Saline-injected animals (i. p.) were used as controls. Animals were culled 4 h after peritonitis induction, and peritoneal lavage and peripheral blood samples were collected. Serum and peritoneal lavage levels of TNF-α as well as TNF-α mRNA expression in peritoneal lavage leukocytes were quantified. Total and differential leukocyte populations were analyzed in peritoneal lavage samples. Lipopolysaccharide-induced increases of both TNF-α mRNA and protein levels were diminished by treatment with either pomegranate leaf hydroalcoholic extract (57 % and 48 % mean reduction, respectively) or sodium diclofenac (41 % and 33 % reduction, respectively). Additionally, the numbers of peritoneal leukocytes, especially neutrophils, were markedly reduced in hydroalcoholic extract-treated rats with acute peritonitis. These results demonstrate that pomegranate leaf extract may be used as an anti-inflammatory drug which suppresses the levels of TNF-α in acute inflammation.

A dosagem de hemoglobina A2 pode ser feitra a partir de amostra de hemolisado com saponina? / Could the saponin hemolysate be used for the hemoglobin A2 dosage?

A dosagem de hemoglobina A2 (HbA2) é comumente feita utilizando-se como amostra solução de hemoglobina (hemolisado clássico com clorofórmio ou tetracloreto de carbono) ou hemolisados com saponina, apesar de os valores de referência para tal determinação terem sido estabelecidos a partir de dados tendo apenas solução de hemoglobina como amostra. Assim, para verificar se há ou näo coerência na utilização indiscriminada daqueles valores de referência para dosagem de HbA2 em amostras de hemolisado com saponina, foi feita a dosagem desta proteína utilizando quatro tipos de hemolisados. Na 1ª etapa da avaliação foram procedidas trinta determinações para fração de HbA2 a partir de hemolisado com saponina e hemolisado clássico com tetracloreto de carbono; na 2ª etapa mais 40 determinações foram analisadas utilizando hemolisado com saponina e com clorofórmio, porém acrescidos de cianeto de potássio (KCN). A análise estatística feita através de teste t pareado na 1ª etapa, revelou uma tendência de que os resultados obtidos a partir de saponina para amostras com níveis normais de HbA2 sejam maiores cerca de 0,42 por cento que aqueles obtidos a partir da solução de hemoglobina (p<0,001). Porém, quando adiciona-se KCN no hemolisado não houve diferença significativa entre o hemolisado clássico (soluçäo de hemoglobina) e o hemolisado com saponina. Deste modo, a utilização de diferentes tipos de hemolisados como amostra (saponina sem KCN ou solução de hemoglobina) podem gerar resultados distintos para a dosagem de HbA2. Determinações de HbA2 feitas especificamente com hemolisado com saponina tendem a superestimar os resultados, quando comparados a valores de referência estabelecidos a partir de hemolisado clássico. Assim, a utilização do hemolisado clássico ou de saponina com KCN são indicados para a dosagem de HbA2 pelo método de eluiçäo eletroforética. O uso de hemolisado apenas com saponina requereria a utilizaçäo de valores de referência específicos. (AU)

It was made the evalution of the hemoglobin A2(Hb A2) determination using four hemolysatetypes. In the 1st. stage of the evalution thirty determinations were proceeded for fraction of Hb A2 startingfrom hemolysate with saponine and starting from carbon tetrachloride, in the 2nd stage more 40determinations were analyzed using hemolysate obtained through saponine and with cloroform, howeveradded with potassium cyanide (KCN). The statistical analysis done through test t in the 1st , it revealed atendency of the results obtained starting from saponine are larger about 0,42% that those obtained startingfrom carbon tetrachloride (p<0,001). However, when KCN is added in the hemolysate there is no significantdifference among the classic hemolisado and with the saponine. Like this, the use of the classic hemolysateor of the saponine with KCN are indicative for Hb A2 quantitative determination for the method of elution. (AU)

Correçäo da interferência dos triglicérideos na dosagem da hemoglobina e na determinaçäo dos índices hematimétricos / Correction of the interference of the triglycerides in the dosage of hemoglobin and hematimetric indexes

A interferência dos triglicerídeos plasmáticos sobre a determinação da hemoglobina foi observada em dosagens acima de 800 mg/dl, provavelmente devido ao aumento da turbidez. Este efeito espúrio leva a falsos resultados no cálculo da HCM e da CHCM. A substituição do plasma lipêmico por soluçäo salina corrige os resultados superestimados da hemoglobina, bem como permite obter resultados mais exatos no cálculo da HCM e da CHCM. (AU)

Plasma triglycerides interference on spectrophotometric hemoglobin determination wasobserved in blood samples with more than 800mg/dl, probably due to the turbidity increase. This spuriouseffect leads to false results in MCM and MCHC determination. Substituting saline solution for thelipaemic plasma corrects the overestimated hemoglobin determination as well as affording more accurateMCH and MCHC values. (AU)

Regional and cellular distribution of serotonin 5-hydroxytryptamine2a receptor mRNA in the nucleus accumbens, olfactory tubercle, and caudate putamen of the rat.

This paper describes the regional and cellular distribution of serotonin 5-hydroxytryptamine2a (5-HT2a) receptor mRNA in (sub)regions of the rat striatum by using in situ hybridization. Our results indicate that 5-HT2a mRNA is distributed heterogeneously in this brain region. Regional densitometry of autoradiograms from striatal sections hybridized with isotope-labeled cRNA probes showed that mRNA levels were highest in the olfactory tubercle, lower in the nucleus accumbens, and lowest in the caudate-putamen. In the nucleus accumbens, the average mRNA levels in the shell were higher than those in the core. These data suggest a particular relevance for the 5-HT2a receptor for olfactory tubercle- and shell-related functions. Therefore, in the nucleus accumbens and the olfactory tubercle, the cellular localization of 5-HT2a mRNA was investigated by determining the colocalization of 5-HT2a mRNA with enkephalin mRNA or dynorphin mRNA. 5-HT2a mRNA was found in enkephalinergic as well as dynorphinergic neurons. Thus, there does not seem to be a differential distribution of this receptor in the output routes of the ventral striatum. In all of the subregions investigated (core, medial shell, and lateral shell of the nucleus accumbens and the olfactory tubercle), only subpopulations of the total enkephalinergic and dynorphinergic populations were found to contain 5-HT2a mRNA. For enkephalin, the percentage colocalization was higher in the lateral shell (61%) compared with the other subregions (38-45%). For dynorphin, the percentage colocalization was higher in the olfactory tubercle (68%) than in the other subregions (34-43%). The differences in (sub)regional mRNA levels and in colocalization with opioids suggest a considerable regional differentiation in the effects of 5-HT2a-mediated neurotransmission in the striatum.