Trypanosuppressive effects of Kolaviron may be associated with down regulation of Trypanothione reductase in Trypanosoma congolense infection.

Trypanothione reductase is a key enzyme that upholds the redox balance in hemoflagellate protozoan parasites such as T. congolense. This study aims at unraveling the potency of Kolaviron against trypanothione reductase in T. congolense infection using Chrysin as standard. The experiment was performed using three different approaches; in silico, in vitro and in vivo. Kolaviron and Chrysin were docked against trypanothione reductase, revealing binding energies (-9.3 and -9.0 kcal/mol) and Ki of 0.211µM and 0.151µM at the active site of trypanothione reductase as evident from the observed strong hydrophobic/hydrogen bond interactions. Parasitized blood was used for parasite isolation and trypanothione reductase activity assay using standard protocol. Real-time PCR (qPCR) assay was implored to monitor expression of trypanothione reductase using primers targeting the 177-bp repeat satellite DNA in T. congolense with SYBR Green to monitor product accumulation. Kolaviron showed IC50 values of 2.64µg/ml with % inhibition of 66.78 compared with Chrysin with IC50 values of 1.86µg/ml and % inhibition of 53.80. In vivo studies following the administration of these compounds orally after 7 days post inoculation resulted in % inhibition of Chrysin (57.67) and Kolaviron (46.90). Equally, Kolaviron relative to Chrysin down regulated the expression trypanothione reductase gene by 1.352 as compared to 3.530 of the infected group, in clear agreement with the earlier inhibition observed at the fine type level. Overall, the findings may have unraveled the Kolaviron potency against Trypanosoma congolense infection in rats.

Trypanosoma brucei brucei infected rats: micronucleated polychromatic erythrocytes.

The emergence of bone marrow micronucleated polychromatic erythrocytes (MN-PCE) in rats experimentally infected with Trypanosoma brucei brucei was examined in order to understand the bone marrow effects in trypanosomiasis infection. Bone marrow was collected for micronucleus assay while blood samples were collected from infected rat for hematological analysis. The results showed evidence of MN-PCE at 12.75 ± 0.65 micronuclei/ 1000 PCE and 9.60 ± 2.95 micronuclei/1000 PCE for rats infected for 21 days and 14 days respectively. The hematology examination revealed changes in packed cell volume, haemoglobin and red blood cells with concomitant increase in parasitemia. This study revealed that the generation of MN-PCE was induced by an acute infection of T. b. brucei in rats and this highlights an important phase in the pathogenesis of the disease that may indicate possible damage to genetic information.