In Vivo Treatment with the Combination of Nitazoxanide and Flubendazole Induces Gluconeogenesis and Protein Catabolism in Taenia crassiceps cysticerci.

PURPOSE: Cysticercosis is the presence of Taenia solium larvae in humans or swines tissues. It is a public health problem related to bad hygienic habits and consumption of infected pork. T. crassiceps is a widely used cysticercosis experimental model. The combination of two effective drugs such as nitazoxanide (NTZ) and flubendazole (FBZ) may potentialize their effect. The aim of this study was to use biochemical analysis to determine the metabolic impact of the combination of NTZ and FBZ on cysticerci inoculated intraperitoneally in mice. METHODS: Balb/c mice intraperitoneally infected with T. crassiceps cysticerci received a single oral dose NTZ/FBZ (50 mg/kg). 24 h after the treatment the cysticerci were removed, frozen and analyzed by high performance liquid chromatography regarding the detection of the following metabolic pathways: glycolysis, gluconeogenesis, homolactic fermentation, tricarboxylic acid cycle, proteins catabolism and fatty acids oxidation. RESULTS: The treatment with the drugs combination induced a statistically significant increase in gluconeogenesis and in protein catabolism when compared to the control groups. CONCLUSION: The drugs combination is potentialized and capable of causing greater metabolic stress than the separate treatment with NTZ or FBZ, showing its potential for an alternative cysticercosis treatment.

In vitro nitazoxanide exposure affects energetic metabolism of Taenia crassiceps.

Nitazoxanide (NTZ) is a broad-spectrum drug used in intestinal infections, but still poorly explored in the treatment of parasitic tissular infections. This study aimed to evaluate the in vitro responses of the energetic metabolism of T. crassiceps cysticerci induced by NTZ. The organic acids of the tricarboxylic acid cycle, products derived from fatty acids oxidation and protein catabolism were analyzed. These acids were quantified after 24 h of in vitro exposure to different NTZ concentrations. A positive control group was performed with albendazole sulfoxide (ABZSO). The significant alterations in citrate, fumarate and malate concentrations showed the NTZ influence in the tricarboxylic acid (TCA) cycle. The non-detection of acetate confirmed that the main mode of action of NTZ is effective against T. crassiceps cysticerci. The statistical differences in fumarate, urea and beta-hydroxybutyrate concentrations showed the NTZ effect on protein catabolism and fatty acid oxidation. Therefore, the main energetic pathways such as the TCA cycle, protein catabolism and fatty acids oxidation were altered after in vitro NTZ exposure. In conclusion, NTZ induced a significant metabolic stress in the parasite indicating that it may be used as an alternative therapeutic choice for cysticercosis treatment. The use of metabolic approaches to establish comparisons between anti parasitic drugs mode of actions is proposed.

Oral nitazoxanide treatment of experimental neurocysticercosis induces gluconeogenesis in Taenia crassiceps cysticerci.

Neurocysticercosis is the most frequent helminthiasis of the central nervous system and is caused by the presence of Taenia solium cysticerci. Nitazoxanide (NTZ) is an antifolate containing the pyrrolopyrimidine-based nucleus that exerts its antiprotozoal activity due to interference with the pyruvate:ferredoxin oxidoreductase (PFOR) enzyme which is essential to anaerobic energy metabolism. The aim of this work was to determine the effect of NTZ on the energetic metabolism of Taenia crassiceps cysticerci intracranially inoculated BALB /c mice. The infected animals were treated with a single oral dose of NTZ 30 days after the inoculation. Analysis of the organic acids was performed through high performance liquid chromatography. Glucose was detected only in the treated groups, alongside with a significant decrease in lactate, pyruvate and oxaloacetate concentrations which indicate an increase in gluconeogenesis. The non-detection of alpha-ketoglutarate indicated the use of the fumarate reductase pathway in all groups. It was possible to confirm the drugs mode of action due to the non-detection of acetate in the treated groups. There was an increase in the fatty acids oxidation. Therefore it was possible to observe that NTZ induces gluconeogenesis as well as the increase of alternative energetic pathways such as fatty acids oxidation in T. crassiceps cysticerci.

In vivo treatment with nitazoxanide induces anaerobic metabolism in experimental intraperitoneal cysticercosis.

Taenia crassiceps cysticerci are used as experimental model to study the host-parasite relationship and treatment of cysticercosis. One of the described mode of actions of nitazoxanide (NTZ) is to block the pyruvate ferredoxine oxidoreductase (PFOR) enzyme which is an essential enzyme to the parasite metabolism. The aim of this study was to determine the in vivo influence of one dosage of NTZ on the energetic metabolism of T. crassiceps cysticerci. Thirty days after the intraperitoneal inoculation of T. crassiceps cysticerci, BALB/c mice were orally treated with 7.5 mg/kg of NTZ. The control group was treated with physiologic solution (NaCl 0.9%). After 24 h, the animals were euthanized and the cysticerci were removed, washed, and processed for biochemical analysis. The organic acids detection occurred through high-performance liquid chromatographic and spectrophotometric analysis. While there was no difference in the glucose dosages, it was possible to observe a significant increase in the lactate concentrations and a decrease in the pyruvate concentrations of the NTZ-treated groups when compared to the control group. Also, there was a decrease in the urea and alpha-ketoglutarate concentrations. This probably occurred due to the impairment of the parasite's PFOR and nitroreductases leading an impairment of the mitochondrial aerobic pathways. In conclusion, the in vivo NTZ treatment leads to an increase in the lactic fermentation and to a decrease in the protein catabolism in T. crassiceps cysticerci.