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Page 6 of 16                                                 Bhatia et al. Hepatoma Res 2018;4:9  I  http://dx.doi.org/10.20517/2394-5079.2018.04

               Superoxide dismutase
               Plasma superoxide dismutase (SOD) activity was assayed according to the method of Kono  based on the
                                                                                             [33]
               ability of SOD to inhibit the reduction of nitroblue tetrazolium (NBT) mediated by superoxide radicals,
               which were produced by photo-oxidation of hydroxylamine hydrochloride. One unit of SOD activity is the
               amount of enzyme required to cause 50% inhibition in optical density. The rate of reduction of NBT complex
               was measured at 560 nm and described as IU/mg protein.


               Assessment of histopathological alterations
               After the completion of the treatment period, mice were euthanized by decapitation and liver tissues were
               removed carefully followed by the immediate fixation of tissue in neutral formalin. Fixed liver tissue samples
               were embedded, sectioned and then stained using hematoxylin and eosin staining and examined for the
               microscopic alterations.


               Statistical analysis
               Data was expressed as mean ± standard deviation (SD). Statistical significance was analyzed using one-way
               analysis of variance followed by least significant difference (LSD) post hoc test.



               RESULTS
               Effect of LycT and/or NDEA on 99mTc-mebrofenin hepatobiliary functional test
               After intravenous administration of  Tc-mebrofenin, NDEA induced a significant delay in the hepatocyte
                                              99m
                                              99m
               uptake, retention and excretion of  Tc-mebrofenin in comparison to control, LycT and LycT + NDEA
               mice [Figure 1A-D]. The hepatic extraction fraction at the end of 60 min of  Tc-mebrofenin injection was
                                                                               99m
               observed to be around 8.9%, 61.9%, 11.5% and 17.8% in case of control, NDEA , LycT and LycT + NDEA
               group respectively [Figure 1E]. Hepatic T  value was observed to be around 5 min in both control and
                                                   peak
               LycT animals [Figure 2A]. However, NDEA administration led to a significant delay in attaining maximum
               activity thus showing a T  value of around 10 min while animals of LycT + NDEA group showed T  value
                                    peak
                                                                                                   peak
               of around 7 min [Figure 2A]. The hepatic T   value in case of control and LycT animals was observed
                                                     1/2 peak
               to be around 7-8 min while it was around 22 min in case of LycT + NDEA group [Figure 2B]. In contrast,
               NDEA animals did not show any T     value which may be due to the extremely slow excretion rate of
               99m Tc-mebrofenin.             1/2peak

               Effect of LycT and/or NDEA on hematological parameters
               Hb
               A significant decline in Hb levels was observed in mice of NDEA group when compared to control and LycT
               (P ≤ 0.001) groups [Table 1]. Likewise, animals of LycT + NDEA group showed a decrease in Hb levels in
               comparison to control and LycT (P ≤ 0.05) mice. However, a significant elevation in Hb levels was observed in
               mice that received LycT along with NDEA treatment when compared to NDEA (P ≤ 0.001) alone injected mice
               [Table 1]. No significant change in Hb levels was noticed between control mice and LycT administered mice.


               RBC
               NDEA administration caused a significant decrease in RBC counts in mice of NDEA group when compared
               to control and LycT (P ≤ 0.001) groups [Table 1]. Also, a decrease in RBC counts was observed in mice of LycT
               + NDEA group when compared to control and LycT (P ≤ 0.01) groups. But the increase in RBC counts was
               observed when NDEA mice were pre-treated with LycT when compared to NDEA (P ≤ 0.01) intoxicated mice
               [Table 1]. The counts of RBC did not differ significantly between mice of control and LycT groups.

               TLC
               TLCs were found to be enhanced in mice of NDEA group when compared to control and LycT (P ≤ 0.001)
               groups [Table 1]. In addition, mice of LycT + NDEA groups also showed elevated total leucocyte counts when
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