RESUMO
Leishmaniasis is a tropical disease caused by protozoan parasites of the genus Leishmania which affects 12 million people worldwide. The discovery of drugs for the treatment of leishmaniasis is a pressing concern in global health programs. The aim of this study aim was to evaluate the leishmanicidal effect of piperine and its derivatives/analogues on Leishmania amazonensis. Our results showed that piperine and phenylamide are active against promastigotes and amastigotes in infected macrophages. Both drugs induced mitochondrial swelling, loose kinetoplast DNA, and led to loss of mitochondrial membrane potential. The promastigote cell cycle was also affected with an increase in the G1 phase cells and a decrease in the S-phase cells, respectively, after piperine and phenylamide treatment. Lipid analysis of promastigotes showed that piperine reduced triglyceride, diacylglycerol, and monoacylglycerol contents, whereas phenylamide only reduced diacylglycerol levels. Both drugs were deemed non toxic to macrophages at 50 µM as assessed by XTT (sodium 2,3,-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino)-carbonyl]-2H-tetrazolium inner salt), Trypan blue exclusion, and phagocytosis assays, whereas low toxicity was noted at concentrations higher than 150 µM. None of the drugs induced nitric oxide (NO) production. By contrast, piperine reduced NO production in activated macrophages. The isobologram analysis showed that piperine and phenylamide acted synergistically on the parasites suggesting that they affect different target mechanisms. These results indicate that piperine and its phenylamide analogue are candidates for development of drugs for cutaneous leishmaniasis treatment.
Assuntos
Alcaloides/uso terapêutico , Benzodioxóis/uso terapêutico , Leishmania/efeitos dos fármacos , Leishmaniose/tratamento farmacológico , Macrófagos/efeitos dos fármacos , Fitoterapia , Piper/química , Piperidinas/uso terapêutico , Alcamidas Poli-Insaturadas/uso terapêutico , Tripanossomicidas/uso terapêutico , Alcaloides/farmacologia , Amidas/farmacologia , Amidas/uso terapêutico , Benzodioxóis/farmacologia , Ciclo Celular/efeitos dos fármacos , Frutas , Glicerídeos/metabolismo , Leishmania/crescimento & desenvolvimento , Leishmania/metabolismo , Leishmaniose/parasitologia , Leishmaniose Cutânea/tratamento farmacológico , Metabolismo dos Lipídeos/efeitos dos fármacos , Macrófagos/parasitologia , Mitocôndrias/efeitos dos fármacos , Óxido Nítrico/biossíntese , Piperidinas/farmacologia , Extratos Vegetais/farmacologia , Extratos Vegetais/uso terapêutico , Alcamidas Poli-Insaturadas/farmacologia , Tripanossomicidas/farmacologiaRESUMO
The current study evaluates the protein and lipid profile of haemolymph of Rhipicephalus (Boophilus) microplus engorged females infected by Metarhizium anisopliae, Beauveria bassiana or Fusarium oxysporum. Ticks were immersed or inoculated with conidial suspension. Haemolymph was collected from the dorsal surface of engorged females. The results showed altered total protein amounts; however, no significant difference was observed on electrophoretic profile among haemolymph samples. In addition, altered lipid profile was detected in haemocyte samples from ticks treated with Beauveria and Metarhizium.
Assuntos
Hemolinfa/química , Lipídeos/química , Fungos Mitospóricos/fisiologia , Proteínas/química , Rhipicephalus/metabolismo , Rhipicephalus/microbiologia , Animais , Eletroforese em Gel de Poliacrilamida , Feminino , Interações Hospedeiro-Patógeno , Esporos FúngicosRESUMO
Lipid transport in arthropods is achieved by highly specialized lipoproteins, which resemble those described in vertebrate blood. Here we describe purification and characterization of the lipid-apolipoprotein complex, lipophorin (Lp), from adults and larvae of the cowpea weevil Callosobruchus maculatus. We also describe the Lp-mediated lipid transfer to developing oocytes. Lps were isolated from homogenates of C. maculatus larvae and adults by potassio bromide gradient and characterized with respect to physicochemical properties and lipid content. The weevil Lp (465 kDa) and larval Lp (585 kDa), with hydrated densities of 1.22 and 1.14 g/mL, contained 34 and 56 percent lipids and 9 and 7 percent carbohydrates, respectively. In both Lps, mannose was the predominant monosaccharide detected by paper chromatography. SDS-PAGE revealed two apolipoproteins in each Lp with molecular masses of 225 kDa (apolipoprotein-I) and 79 kDa (apolipoprotein-II). The lipids were extracted and analyzed by thin-layer chromatography. The major phospholipids found were phosphatidylserine, phosphatidylcholine and phosphatidylethanolamine in adult Lp, and phosphatidylcholine, phosphatidylethanolamine and sphingomyelin in larval Lp. Hydrocarbons, fatty acids and triacylglycerol were the major neutral lipids found in both Lps. Lps labeled in the protein moiety with radioactive iodine (125I-iodine) or in the lipid moiety with fluorescent lipids revealed direct evidence of endocytic uptake of Lps in live oocytes of C. maculatus.
Assuntos
Animais , Feminino , Hidrocarbonetos/análise , Metabolismo dos Lipídeos/fisiologia , Lipoproteínas/química , Oócitos/crescimento & desenvolvimento , Fosfolipídeos/química , Gorgulhos/química , Apolipoproteínas/química , Apolipoproteínas/isolamento & purificação , Apolipoproteínas/metabolismo , Transporte Biológico , Endocitose/fisiologia , Lipoproteínas/isolamento & purificação , Lipoproteínas/metabolismo , Oócitos/metabolismo , Oogênese/fisiologia , Fosfolipídeos/isolamento & purificação , Fosfolipídeos/metabolismo , Gorgulhos/metabolismoRESUMO
Lipid transport in arthropods is achieved by highly specialized lipoproteins, which resemble those described in vertebrate blood. Here we describe purification and characterization of the lipid-apolipoprotein complex, lipophorin (Lp), from adults and larvae of the cowpea weevil Callosobruchus maculatus. We also describe the Lp-mediated lipid transfer to developing oocytes. Lps were isolated from homogenates of C. maculatus larvae and adults by potassio bromide gradient and characterized with respect to physicochemical properties and lipid content. The weevil Lp (465 kDa) and larval Lp (585 kDa), with hydrated densities of 1.22 and 1.14 g/mL, contained 34 and 56% lipids and 9 and 7% carbohydrates, respectively. In both Lps, mannose was the predominant monosaccharide detected by paper chromatography. SDS-PAGE revealed two apolipoproteins in each Lp with molecular masses of 225 kDa (apolipoprotein-I) and 79 kDa (apolipoprotein-II). The lipids were extracted and analyzed by thin-layer chromatography. The major phospholipids found were phosphatidylserine, phosphatidylcholine and phosphatidylethanolamine in adult Lp, and phosphatidylcholine, phosphatidylethanolamine and sphingomyelin in larval Lp. Hydrocarbons, fatty acids and triacylglycerol were the major neutral lipids found in both Lps. Lps labeled in the protein moiety with radioactive iodine (125I-iodine) or in the lipid moiety with fluorescent lipids revealed direct evidence of endocytic uptake of Lps in live oocytes of C. maculatus.
Assuntos
Hidrocarbonetos/análise , Metabolismo dos Lipídeos/fisiologia , Lipoproteínas/química , Oócitos/crescimento & desenvolvimento , Fosfolipídeos/química , Gorgulhos/química , Animais , Apolipoproteínas/química , Apolipoproteínas/isolamento & purificação , Apolipoproteínas/metabolismo , Transporte Biológico , Endocitose/fisiologia , Feminino , Lipoproteínas/isolamento & purificação , Lipoproteínas/metabolismo , Oócitos/metabolismo , Oogênese/fisiologia , Fosfolipídeos/isolamento & purificação , Fosfolipídeos/metabolismo , Gorgulhos/metabolismoRESUMO
We have characterized the serum lipoprotein profile and localized the serum paraoxonase activity of pacu, Piaractus mesopotamicus, a tropical fish species. The total lipoprotein profile of pacu serum obtained after KBr density ultracentrifugation shows the predominance of HDL (1.1267 g/mL). SDS-PAGE electrophoresis revealed a negligible amount of LDL. Pacu HDL was purified by gel filtration column on HPLC, and its molecular mass was estimated to be 246 kDa. Protein composition was 35%, and comprised four protein components with molecular masses of 45, 38, 25 and 12.5 kDa. Lipids represent 58% of total HDL, comprising 40% neutral lipids and 18% phospholipids by weight. The HDL contains 7% of carbohydrates, and mannose was the only sugar detected by paper chromatography in HDL hydrolysates. HDL-containing fractions showed the major paraoxonase activity. Purification of HDL resulted in a 23-fold enrichment of this activity. This is the first experimental evidence demonstrating the association of paraoxonase activity with a HDL in fish.