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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