Polyphenolic characterisation of plant mixture (Lisosan® Reduction) and its hypocholesterolaemic effect in high fat diet-fed mice

<p>Lisosan® Reduction is a plant mixture produced from powder of fermented <i>Triticum aestivum</i> (Lisosan® G)<i>, Desmodium adscendens, Malus domestica, Picrorhiza kurroa</i> and <i>Hordeum vulgare.</i> The aim of this study was to characterise the phenolic profile of Lisosan® Reduction and to evaluate the effects of aqueous extract on mice fed a high fat diet (HFD). Syringic acid, trans sinapic acid and neochlorogenic acid were identified by HPLC-DAD to be the dominant polyphenols of Lisosan® Reduction, followed by vitexin, trans p-coumeric acid and trans ferulic acid. Mice treated with aqueous extract of Lisosan® Reduction (60 mg/kg b.w.) showed a significant decrease of serum cholesterol, glucose and triglycerides level and a significant increase of CYP7A1 gene expression, compared to HFD group.</p

Quantitative real-time RT-PCR analysis of GPx-1, SOD2, CAT and HO-1 expression in EPCs after incubation with lysates in absence (panel A) or in presence of oxidative stress induced by H₂O₂ (panel B).

<p>The bars represent mean±SE fold increase in transcript expression relative to untreated cells (CNT); n≥3. *p<0.05, **p<0.01 vs control (CNT); # p<0.05, ## p<0.01 vs CNT + H₂O₂.</p

Assessment of Nrf-2 translocation into the nucleus by fluorescence microscopy.

<p>Untreated EPCs (panel A); untreated EPCs exposed to oxidative stress induced by H₂O₂ (panel C). Nucleus localization of Nrf-2 (green) was present in both EPCs pre-treated with LG (panel B), and also in EPCs exposed to oxidative stress induced by H₂O₂ and pre-treated with LG (panel D). DAPI was used for nuclei staining (blue).</p

Effects of Lisosan G (LG) and LJ lysate (LJ) on viability of EPCs in absence (panel A) or presence of oxidative stress induced by H₂O₂ (panel B).

<p>Data are expressed as means ±SE; n≥3. *p<0.05, **p<0.01, ****p<0.001 vs control (CNT); #p<0.05 vs CNT + H₂O₂.</p

Effects of Lisosan G (LG) and LJ lysate (LJ) on EPCs adhesion in absence (panel A) or presence of oxidative stress induced by H₂O₂ (panel B).

<p>Data are expressed as means ±SE; n≥3. **p<0.01, ****p<0.001 vs control (CNT); # p<0.05, ## p<0.01, #### p<0.001 vs CNT + H₂O₂.</p

Effects of LG and LJ on ROS production in absence (panel A) or in presence of oxidative stress induced by H₂O₂ (panel B).

<p>Data are expressed as means ± SE; n≥3. # p<0.05, ## p<0.01 vs CNT + H₂O₂.</p

Effects of Lisosan G (LG) and LJ lysate (LJ) on EPCs senescence in absence (panel A) or presence of oxidative stress induced by H₂O₂ (panel B).

<p>Data are expressed as means ±SE; n≥3. *p<0.05, **p<0.01 vs control (CNT); # p<0.05, ## p<0.01 vs CNT + H₂O₂.</p

Representative immunofluorescence of retinal sections showing the expression pattern of GLAST following ischemia.

<p>Glast immunoreactivity in the ischemic retina after 150 min of reperfusion (ISCH/REP) shows no evident changes in the expression and distribution of the glial transporter compared to the contralateral non-ischemic retina (CTL). (GCL) ganglion cell layer (IPL), inner plexiform layer, (INL) inner nuclear layer, (OPL) outer plexiform layer, (ONL) outer nuclear layer.</p

Modulation of [³H]-D-Aspartate release and uptake in synaptosomes from ischemic retina.

<p>(A) <i>Ischemia reduced [³H]-D-Asp uptake in highly pure nerve ending.</i> [³H]-D-Asp uptake into synaptosomes was measured in animals subjected to retinal ischemia for 50 min in the right eye and reperfused for 150 min (dashed line). The contralateral eye (solid line) was used as control. Data are expressed as mean ± S.E.M. (n = 3 or 4 animals per group). (B) <i>[³H]-D-Asp release in retinal synaptosomes.</i> Animals were subjected to retinal ischemia for 50 min in the right eye and reperfusion was allowed for 150 min. Release was measured in the ischemic (white histograms), contralateral (grey) and naïve retinas (black). All results are expressed as percentage of the corresponding KCl-evoked release (15 mM). Omission of Ca²⁺ produces a reduction of [³H]-D-Asp release of about 40% in all groups, while DL-TBOA reduces by about 60% the release in naïve and contralateral samples, but not in the ischemic retina. *p<0.05 vs control.</p

Effects of retinal ischemia/reperfusion on the expression and distribution of glutamate transporter GLT-1.

<p>GLT-1 immunoreactivity increases in the retina of rats subjected to 50 min of ischemia followed by 150 min of reperfusion (ISCH/REP) as compared to the control, non-ischemic retina (CTL). An increased number of GLT-1 positive cell bodies is evident in the ONL and INL of retinas subjected to ischemia (ISCH/REP). (GCL) ganglion cell layer (IPL), inner plexiform layer, (INL) inner nuclear layer, (OPL) outer plexiform layer, (ONL) outer nuclear layer.</p