Following deparaffination, areas were stained with hematoxylin and eosin (H&E), mounted in DPX, and imaged under a bright-field microscope (Carl Zeiss AxioVision, Oberkochen, Germany). Genotoxicity evaluation To evaluate genotoxicity caused by Disarib at the effective dose, micronucleus assay was performed32C34. of Disarib to ABT199, the only FDA approved BCL2 inhibitor revealed that Disarib is as potent as ABT199. Recent studies using mice revealed that Disarib did not invoke significant side effects in mice. In the present study, we have investigated the acute toxicity of Disarib in Wistar rats. The bioavailability studies following exposure of Disarib in Wistar rats revealed its maximum availability in serum at 24?h following oral administration. Acute toxicity analysis revealed that even a dose as high as 2000?mg/kg of Disarib did not cause significant toxicity in rats. There was no significant variation in blood parameters or kidney and liver functions following administration of Disarib. Histological analysis of different tissues from Disarib treated groups revealed standard architecture with no observable cellular damage. Importantly, exposure to Diasrib did not result in 3,3′-Diindolylmethane genotoxicity as determined by micronucleus assay. Further, solubility assays revealed that besides DMSO, Disarib is also soluble in alcohol. While the high acidic condition can increase the solubility of Disarib, 3,3′-Diindolylmethane even a lower percentage of alcohol with acidic conditions can improve its solubility. Thus, the toxicological profile in the current study revealed no significant side effects when Disarib was administered orally to rats. not significant). Rabbit Polyclonal to NT5E Haematological studies show normal blood parameters following Disarib treatment in rats Blood was collected by heart puncture from treated and control Wistar rats after the 14th day of administration of Disarib in anticoagulant coated vials, and total blood count was evaluated. Results showed no significant difference in the total number of RBC and HGB content between control and treated groups, and the values were within the normal range (Fig.?5)28C30. The decrease in platelet count upon higher doses is commonly reported for most BCL2 inhibitors4,19. Although a marginal variation in platelet count was observed in the case of 1000?mg/kg b. wt. Disarib treated group; such a difference was not observed in lower and higher dose treated groups (100 and 2000?mg/kg, respectively) (Fig.?5). Therefore, these results suggested that Disarib treatment in rats did not result in dose-limiting toxicity such as thrombocytopenia (Fig.?5). The analysis also revealed that there was no significant difference in WBC counts between control and treated groups. Further, no significant difference was observed between the neutrophils and lymphocytes count in control and treated animals (Fig.?5). Although there were some variations among different groups and animals within the group, all the values were within the normal physiological range (Fig.?5)28,30. Analyses of PCV, MCV, MCH, MCHC, eosinophils, and monocytes suggested no toxic effects caused by Disarib as all these parameters were comparable to that of the control group (Table ?(Table11). Open in a separate window Figure 5 Blood parameter analysis following oral administration of Disarib in rats. Analysis of red blood corpuscles (RBC), white blood corpuscle (WBC), haemoglobin (HGB), Platelets, Neutrophils, Lymphocytes count among control and 3,3′-Diindolylmethane Disarib treated groups (100, 1000, 2000?mg/kg b. wt) after 14?days of dose administration. Error bars denote mean?+?SEM (not significant). Table 1 Analyses of blood parameters after 14?days of oral administration of Disarib in Wistar rats. packed cell volume, mean cell volume, mean corpuscular haemoglobin, mean corpuscular haemoglobin concentration. Administration of Disarib did not affect hepatic or renal functions After acute dose administration of Disarib in rats, serum was collected 14?days post treatment and analyzed for factors that indicate kidney and liver functions. Hepatic function tests for SGPT, ALP, SGOT, total protein, Albumin, and Bilirubin suggested that Disarib did not cause any adverse effect on liver function (Fig.?6A). Although, ALP levels observed were higher in 100 and 3,3′-Diindolylmethane 1000?mg/kg Disarib treated groups compared to control, the increase was limited and was not significant. Moreover, the group treated with 2000?mg/kg Disarib showed an ALP level similar to that of the control (Fig.?6A)28,30,31. Consistent with this, the level of SGOT was marginally higher in the maximal dose treated group; however, the increase was within the normal range and was not significant compared to control28 (Fig.?6A). Renal function analysis showed that BUN levels, Creatinine, Phosphorous, and Uric acid did not have any appreciable variation between treated and control groups (Fig.?6B). Thus, the above results suggested that.
