ABSTRACT
Hypnophilin and panepoxydone, terpenoids isolated from Lentinus strigosus, have significant inhibitory activity onTrypanosoma cruzi trypanothione reductase (TR). Although they have similar TR inhibitory activity at 10 μg/mL (40.3 μM and 47.6 μM for hypnophilin and panepoxydone, respectively; ~100 percent), hypnophilin has a slightly greater inhibitory activity (~71 percent) on T. cruzi amastigote (AMA) growth in vitro as well as on in vitro phytohemagglutinin (PHA)-induced peripheral blood mononuclear (PBMC) proliferation (~70 percent) compared to panepoxydone (69 percent AMA inhibition and 91 percent PBMC inhibition). Hypnophilin and panepoxydone at 1.25 μg/mL had 67 percent inhibitory activity onLeishmania (Leishmania) amazonensis amastigote-like (AMA-like) growth in vitro. The panepoxydone activity was accompanied by a significant inhibitory effect on PHA-induced PBMC proliferation, suggesting a cytotoxic action. Moreover, incubation of human PBMC with panepoxydone reduced the percentage of CD16+ and CD14+ cells and down-regulated CD19+, CD4+ and CD8+ cells, while hypnophilin did not alter any of the phenotypes analyzed. These data indicate that hypnophilin may be considered to be a prototype for the design of drugs for the chemotherapy of diseases caused by Trypanosomatidae.
Subject(s)
Humans , Antiprotozoal Agents/pharmacology , Bridged Bicyclo Compounds, Heterocyclic/pharmacology , Leishmania/drug effects , Lentinula/chemistry , Plant Extracts/pharmacology , Sesquiterpenes/pharmacology , Trypanosoma cruzi/drug effects , Antigens, CD/drug effects , Bridged Bicyclo Compounds, Heterocyclic/isolation & purification , Cell Proliferation/drug effects , Drug Design , Leukocytes, Mononuclear/drug effects , Sesquiterpenes/isolation & purificationABSTRACT
Estudou-se atividade antineoplásica de um produto natural isolado de Alomia myriadenia (miriadenolídeo) no modelo do tumor de Ehrlich em camundongos. Dezoito fêmeas de camundongo Swiss foram inoculadas com 2x10(7) células viáveis de tumor de Ehrlich via intraperitoneal (0,3ml) e posteriormente distribuídas aleatoriamente em três grupos que receberam: grupo I (controle) - 0,3ml de solução de Hanks; grupo II - 31µg/kg de miriadenolídeo; e grupo III - 139µg/kg de miriadenolídeo. No oitavo dia de experimento, foram realizados exames hematológicos e perfil protéico sérico eletroforético. Coletou-se todo o líquido ascítico para avaliação do volume, aparência, pH, contagem de células viáveis e inviáveis, realização de esfregaços para contagem de células claras e escuras, leucócitos e avaliação das regiões organizadoras de nucléolos argentafins (AgNORs). Foram realizados exames macro e microscópicos do baço, fígado e rins e aspirado o conteúdo da medula óssea dos fêmures direito e esquerdo de cada animal para avaliação da relação mielóide:eritróide. Não houve diferença significativa no volume, pH, contagem de células viáveis e inviáveis entre os três grupos estudados, observando-se valores de 17,6 x 10(4) células tumorais viáveis no grupo III, 27,7 x 10(4) no grupo II e 21,1 x 10(4) no grupo I. As AgNORs apresentaram-se pequenas, com distribuição difusa e incontáveis no grupo I, e em menor quantidade no grupo III. Os animais do grupo III apresentaram a menor concentração protéica total sérica (4,7g/dl) (P<0,05) quando comparados com os do grupo II (5,3g/dl) e do grupo I (5,1g/dl). Os valores de albumina foram semelhantes nos três grupos (2,6g/dl), e as globulinas totais foram maiores (P<0,05) no grupo II (2,71g/dl) quando comparadas com os valores médios do grupo III (2,11g/dl) e semelhantes ao grupo I (2,43g/dl). Não houve diferença entre alfa e beta globulinas entre os três grupos estudados, porém as gamaglobulinas foram maiores...
Antitumoral activity of a natural product of Alomia myriadenia (myriadenolide) in Ehrlich tumor in mice was studied. Eighteen Swiss female mice were intra-peritoneal inoculated 2x10(7) viable cells of Ehrlich Tumor (0.3ml) and randomly distributed in three groups receiving via intra-peritoneal on the 3rd and 5th day post-inoculation the following treatments: group I (control) - 0.3ml Hanks solution; group II: 31µg/kg myriadenolide; and group III: 139µg/kg myriadenolide. On the eighth day of the experiment blood profile and protein serum electrophoresis were performed. All ascitic liquid was collected to evaluate the volume and pH; to observe the aspect; to count viable and no viable cells, dark and clear cells, leukocytes and nucleolar organizer regions (NORs). Macro and microscopic exams were performed and bone marrow was aspirated from right and left femurs of each animal to evaluate myeloid:erythroid ratio. It was not observed difference in volume, pH, counts viable and no viable cells in the groups, although group III showed smaller number of viable tumoral cells (17.6 x 10(4)) when compared to the group II (27.7 x 10(4)) and group I (21.1 x 10(4)). The investigation of NORs to evaluate the proliferative capacity of tumoral cells after myriadenolide treatment showed that cells were smaller, uncountable and with diffuse distribution in group I. They were in lower quantity in group III. These results suggest that myriadenolide in dose 139µg/kg (group III) delay the tumoral growing and, probably, cell proliferation. The animals of group III showed lower value of total protein (4.7g/dl) (P<0.05) when compared to animals from group II (5.3g/dl) and group I (5.1g/dl). The values of albumin were similar in all groups (2.6g/dl) and total globulin was higher (P<0.05) in group II (2.71g/dl) when compared to mean values of group III (2.11g/dl) and similar to group I (2.43g/dl). The decrease of total protein in group III occurred...
Subject(s)
Animals , Female , Asteraceae/adverse effects , Carcinoma, Ehrlich Tumor/diagnosis , Carcinoma, Ehrlich Tumor/prevention & control , Electrophoresis/methods , MiceABSTRACT
A procedure is described for the rapid determination of the intra-erythrocyte concentration of 6-mercaptopurine (6-MP) and its metabolites, 6-thioguanine nucleotides (6-TGN) and 6-methylmercaptopurine (6-MMP). Erythrocytes (8 x 10(8) cells) in 350 æl Hanks solution containing 7.5 mg dithiothreitol were treated with 50 æl 70 percent perchloric acid. The precipitate was removed by centrifugation (13,000 g) and the supernatant hydrolyzed at 100§C for 45 min. After cooling, 100 æl was analyzed directly by HPLC using a Radialpack Resolve C18 column eluted with methanol-water (7.5:92.5, v/v) containing 100 mM triethylamine. 6-TG, 6-MP and the hydrolysis product of 6-MMP, 4-amino-5-(methylthio)carbonyl imidazole, were monitored at 342, 322 and 303 nm using a Shimadzu SPD-M10A diode array UV detector. The analytes eluted at 5.3, 6.0 and 10.2 min, respectively. The calibration curves were linear (rý > 0.998), and the analytical recoveries were 73.2 percent for 6-TG, 119.1 percent for 6-MP and 97.4 percent for 6-MMP. The intra- and inter-assay variations were highest for 6-MP (9.6 and 14.3 percent, respectively). The lowest detectable concentrations were 3, 3 and 25 pmol/8 x 10(8) erythrocytes for 6-TG, 6-MP and 6-MMP, respectively. The quantification limits (coefficients of variation <15 percent) were 8, 10 and 70 pmol/8 x 10(8) erythrocytes for 6-TG, 6-MP and 6-MMP, respectively. The method was applied to the analysis of 183 samples from 36 children under chemotherapy for acute lymphoblastic leukemia. The concentrations of the metabolites in the red cells of the patients ranged from 0 to 1934 pmol/8 x 10(8) erythrocytes for 6-TGN, and from 0 to 105.8 and 0 to 45.9 nmol/8 x 10(8) erythrocytes for 6-MP and 6-MMP, respectively. The procedure gave results that were in agreement with those obtained with other methods designed to detect cases of non-compliance with treatment, including patient interviews and medical evaluation, among others, demonstrating its applicability to monitoring the treatment of leukemic children.