Ellagic acid attenuates beryllium sulphate-induced oxidative stress and histopathological alterations of spleen in rats

Figures & data

Figure 1. Change in the body weight (BW) during the experiment/Body weights were measured every day. Data are represented as mean ± SD (n = 10). Data were analysed by one way analysis of variance (ANOVA). *p < 0.05: Significant difference in comparison with the control group. ^#p < 0.05: Significant difference in comparison with the BeSO₄-treated group.

Table 1. Effect of EA on the spleen weight in BeSO₄-intoxicant rats.

Groups	Final body weight (g)	Spleen weight (g)	Relative spleen weight to the body weight (%)
Control	502.81 ± 5.13	0.216 ± 0.003	0.042 ± 0.009
BeSO4	499.79 ± 3.90	0.255 ± 0.003*	0.051 ± 0.005*
EA 100	499.70 ± 4.77	0.225 ± 0.001^#	0.045 ± 0.004^#
EA 300	502.03 ± 3.40	0.215 ± 0.002^#	0.043 ± 0.004^#

After the final administration, animals were sacrificed by decapitation and spleen tissue samples were weighted and the weight of the spleens relative to the body weight was calculated.

Data are represented as mean ± SD (n = 10). Data were analysed by one way analysis of variance (ANOVA).

*p < 0.05: Significant difference in comparison with the control group.

^#p < 0.05: Significant difference in comparison with the BeSO₄-treated group.

Table 2. Effect of EA on hematological parameters in BeSO₄-intoxicant rats.

	Control	BeSO4	EA 100	EA 300
WBC (10^9/L)	6.74 ± 0.20	11.02 ± 1.31*	9.22 ± 0.63^#	7.03 ± 0.82^#
Neu (10^9/L)	1.14 ± 0.11	2.45 ± 0.42*	1.66 ± 0.13^#	1.08 ± 0.25^#
Lym (10^9/L)	3.80 ± 0.83	9.64 ± 1.99*	6.37 ± 1.28^#	3.92 ± 0.96^#
PLT (10^9/L)	868.4 ± 43.2	1408 ± 77.57*	1194 ± 28.90^#	872.4 ± 86.4^#

Blood sample was collected from the abdominal aorta. The whole blood cell count was used for hematological analysis.

WBC: white blood cell count; Neu: neutrophil count; Lym: lymphocyte count; PLT: platelet count.

Data are represented as mean ± SD (n = 5). Data were analysed by one way analysis of variance (ANOVA).

*p < 0.05: Significant difference in comparison with the control group.

^#p < 0.05: Significant difference in comparison with the BeSO₄-treated group.

Figure 2. Effect of EA on splenocyte proliferation in BeSO₄-intoxicant rats/Cultured splenocytes were suspended in RPMI-1640 complete medium containing 12% FBS, 100 U/mL penicillin and 100 µg/mL streptomycin, seeded into 96-well plates at 5 × 10₄ cells/well and incubated with or without Con A (A) or LPS (B). Data are represented as mean ± SD (n = 5). Data were analysed by one way analysis of variance (ANOVA). *p < 0.05: Significant difference in comparison with the control group. ^#p < 0.05: Significant difference in comparison with the BeSO₄-treated group.

Figure 3. Effect of EA on GSH content, SOD activity and MDA level in BeSO₄-intoxicant rats/GSH content was measured fluorometrically using orthophthalaldehyde (A). The SOD activity was measured using commercial ELISA kits (B). The level of MDA was measured using MDA assay kit (C). Data are represented as mean ± SD (n = 5). Data were analysed by one way analysis of variance (ANOVA). *p < 0.05: Significant difference in comparison with the control group. ^#p < 0.05: Significant difference in comparison with the BeSO₄-treated group.

Figure 4. Effects of EA on apoptosis in BeSO₄-intoxicant rats/Relative mRNA expression levels of Bax, Bcl-2, Caspase-3, and PARP are shown in Figure 3 (A,B,C,D). The expression levels of Bax, Bcl-2, Caspase-3, and PARP proteins were detected by western blotting are shown in Figure 4 E. Data are represented as mean ± SD (n = 5). Data were analysed by one way analysis of variance (ANOVA). *p < 0.05: Significant difference in comparison with the control group. ^#p < 0.05: Significant difference in comparison with the BeSO₄-treated group.

Figure 5. Histopathological observations showing effects of EA on BeSO₄-induced changes in spleen/Spleen tissues were subjected to H&E staining. Microscopic images were taken at a magnification of 40 × (A–D), 200 × (E–H) and 400 × (I–L) for the spleen. White arrow: thickening of the marginal area; Black arrow: localized central dilatation of the spleen nodules; Yellow arrow: red pulp with blood stasis; Red arrow: nuclear fragmentation.

Figure 6. Proposed mechanism of ellagic acid against BeSO₄-induced splenic toxicity.