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Sharma et al. Zinc supplementation prevents against LPS induced neurotoxicity in rats

of Humanson. All brain tissue sections were fixed Prenatally LPS exposed pups of both genders showed
[48]
in 10% neutral buffered formalin. The tissues were significant (P < 0.05) increase (2-fold) in the number
processed and embedded in paraffin and sectioned of escaped trials when compared to control pups.
at 4 μm thickness. The sections were stained with However, number of escaped trials were significantly
hematoxylin and eosin (HE). The dried stained slides (P < 0.05) decreased in male (35.29%) and female
were seen under the Leica microscope. Images (41.67%) pups following zinc supplementation to LPS
were captured at 10× and 40× and auto corrected in treated mothers when compared to prenatally LPS
Microsoft office 2010 to enhance image clarity. treated pups.

Statistical analysis Effect of prenatal zinc supplementation on
Statistical analysis was done by one way ANOVA neurotransmitter levels in pups
followed by LSD with post-hoc multiple pairwise Present study relates the deficit in locomotor activity
comparisons between genders and different groups to and short term memory with neurotransmitter levels
estimate whether the differences between the mean which were measured using HPLC as shown in
values of groups are statistically significant or not. Table 1. Prenatal LPS exposure significantly (P < 0.05)
Results were taken significant at P ≤ 0.05. decreased the level of dopamine in male (38.37%) and
female (41.70%) pups when compared to control pups.
RESULTS In case of pups from zinc supplemented mothers a
significant (P < 0.05) improvement in dopamine level
Prenatal zinc supplementation improves LPS was observed (male 52.2%) and (female 13.37%)
induced neurobehavioral deficits in pups when compared to prenatally LPS treated pups.
Total locomotor activity was assessed in prenatally
LPS treated male and female pups on actophotometer. Prenatal LPS exposure significantly (P < 0.05) increased
Prenatal LPS exposure significantly (P < 0.05) the level of norepinephrine (NE) in male (78.21%)
decreased total locomotor activity in male (31.59%) and female (24.42%) pups when compared to control
as well as female (30.17%) pups when compared pups. However, NE levels were found to decrease
to control pups. Zinc supplememtaion to female rats significantly (P < 0.05) in male (27.98%) as well as
throughout pregnancy improved locomotor activity female pups (14.42%) following zinc supplementation
in case of female pups (21.60%) [Figure 1]. In case to LPS treated mothers when compared to prenatally
of rotarod test a significantly (P < 0.05) decreased LPS treated pups.
mean fall off time was observed in (54.30%) male and
(65.93%) female pups in comparison to control pups. Prenatal LPS exposure also significantly (P < 0.05)
Zinc supplementation however significanty increased decreased the level of 5-HT in male (30.58%) and
the mean fall off time in pups of both sexes. female (55.76%) pups when compared to control pups.
Whereas, 5-HT levels were significantly (P < 0.05)
Spatial memory was assessed in prenatally LPS improved with (14.40%) increase in male and female
treated male and female pups using a EPM as well as (60.86%) pups following zinc supplementation to LPS
active avoidance test as shown in Figure 2. Time spent treated mothers when compared to prenatally LPS
in closed arm was significantly (P < 0.05) decreased treated pups.
in case of prenatally LPS treated male pups (66.23%)
and female pups (6-fold) when compared to control Effect of prenatal zinc supplementation on
pups. However, following zinc supplementation time oxidative stress markers (NO levels, MDA level
spent in the closed arm was significantly (P < 0.05) (lipid peroxidation) and antioxidant enzymes
increased in male (14.84%) and female (47.26%) pups (GSH, SOD and catalase)
when compared to LPS treated pups. As seen from Table 2 a significant (P < 0.05) elevation
Table 1: Effect of prenatal zinc supplementation on neurotransmitter levels in mid brain of prenatally LPS treated
male and female pups
Dopamine (ng/mg tissue) Serotonin (ng/mg tissue) Norepinephrine
Group
Male Female Male Female Male Female
Control 0.645 + 0.050 0.885 + 0.040 0.340 + 0.030 0.260 + 0.007 3.81 + 0.08 5.69 + 0.13
LPS 0.398 + 0.030 * 0.516 + 0.060 * 0.236 + 0.000 * 0.115 + 0.005 * 6.79 + 0.08 * 7.07 + 0.12 *
0.605 + 0.050 # 0.585 + 0.030 # 0.270 + 0.010 # 0.185 + 0.005 # 4.89 + 0.08 # 6.05 + 0.11 #
LPS + ZnSO 4 *# # *# # *# *#
0.748 + 0.020 0.901 + 0.010 0.313 + 0.010 0.245 + 0.005 3.08 + 0.08 5.05 + 0.11
ZnSO 4
Values are expressed as mean + SD; n = 5/group. *P < 0.05 vs. control group, P < 0.05 vs. prenatally LPS treated group. LPS:
#
lipopolysaccharide
Neuroimmunology and Neuroinflammation ¦ Volume 4 ¦ March 21, 2017 37