The effect of DBME on liver parameters (SOD, CAT, GST, GSH) in iron overloaded mice.

<p>Values are mean ± SD of six observations.</p><p>^a<i>p<</i>0.05, ^b<i>p<</i>0.01and ^c<i>p<</i>0.001 significant difference from normal mice (B) group</p><p>^d<i>p<</i>0.05, ^e<i>p<</i>0.01 and ^f<i>p<</i>0.001 significant difference from iron overloaded (C) group</p><p>The effect of DBME on liver parameters (SOD, CAT, GST, GSH) in iron overloaded mice.</p

The changes in ALAT, ASAT, ALP and Bilirubin levels after oral therapy with PNME.

<p>The changes in ALAT, ASAT, ALP and Bilirubin levels after oral therapy with PNME.</p

Phytochemical analysis of PNME.

<p>Phytochemical analysis of PNME.</p

Metal chelation potential of PNME.

<p><b>A.</b> Iron chelation assay, <b>B.</b> Inhibition of lipid peroxidation, <b>C.</b> DNA protection assay. Agarose gel showing bands of supercoiled (SC) and open circular (OC) forms of pUC-18 DNA, and the graph denotes the % protection of supercoiled DNA by PNME. The results are the mean ± S.D. (n = 6). **p < 0.01 and ***p < 0.001 vs. control.</p

IC₅₀ values of purpurin, catechin, tannic acid, reserpine, methyl gallate, rutin and DBME for <i>in vitro</i> iron chelation and cytotoxicity against WI-38 cells.

<p># All the IC₅₀ values are determined in μg/ml. Data expressed as mean ± S.D (n = 6).</p><p>IC₅₀ values of purpurin, catechin, tannic acid, reserpine, methyl gallate, rutin and DBME for <i>in vitro</i> iron chelation and cytotoxicity against WI-38 cells.</p

RNS scavenging activity of PNME.

<p><b>A.</b> Nitric oxide inhibition, <b>B.</b> Peroxynitrite radical scavenging. The results are the mean ± S.D. of six parallel measurements. ***p < 0.001 vs. control.</p

Quantification of phytochemicals of DBME.

<p>Quantification of phytochemicals of DBME.</p

Microscopic observation of mouse liver sections stained with hematoxylin and eosin at 400x.

<p><b>A</b>. Liver section from control mice with normal cytoarchitecture. <b>B</b>. Iron-overloaded (iron dextran, 100 mg/kg b.w.) liver section shows degeneration of cellular boundaries, fatty ballooning deterioration, inflammation <b>(I)</b>, and necrosis <b>(N)</b>. <b>C</b>. Liver section of S50 shows better cytostructure with minor inflammation <b>(PI)</b>. <b>D</b>. Liver section of S100. <b>E</b>. Liver section of S200. <b>F</b>. Desirox-treated liver section shows a reduced necrotic area.</p

Photomicrograph of mice liver sections (Masson’s Trichrome staining) ×100.

<p>(A) Control mice liver shows normal cellular integrity with no fibrosis. (B) Iron-intoxicated (iron dextran, 100 mg/kg b.w.) liver section with elongated fibrous septa and accumulation of collagen (Blue). (C) Liver section treated with iron dextran + 50 mg/kg b.w. DBME shows slight fibrosis. (D) Liver section treated with iron dextran + 100 mg/kg b.w. DBME shows improved histology. (E) Liver section treated with iron dextran + 200 mg/kg b.w. DBME. (F) Liver section treated with iron dextran + 20 mg/kg b.w. desirox shows nearly negligible accumulation of collagen and healthy liver. S100 and S200 show reduced collagen deposition, fibrous septum and necrotic cells in periportal veins indicating a trend of restoration of normal cellular integrity.</p

Microscopic observation of liver sections stained with Masson’s trichrome at 100x.

<p>A. Liver section from control mice showing no indication of fibrosis. <b>B</b>. Liver section of group C exhibiting significant accumulation of collagen (Blue) with elongated fibrous septa. <b>C</b>. Liver section from the S50 group shows negligible fibrosis. <b>D</b>. Liver section from the S100 group and S200 group (E) depicted similar liver architecture improvement as Desirox-treated group (F).</p