Effect of Lisosan G on activity of antioxidant and phase II drug-metabolizing enzymes.

<p>NAD(P)H:quinone oxidoreductase (control value: 53.23±2.5 nmol/min/mg prot) (A); Heme oxygenase-1 (control value: 9.57±3.86 pmol/min/mg prot) (B); Glutathione-S-transferase (control value: 239.5±12.12 nmol/min/mg prot) (C); Catalase (control value: 147.41±11.1 nmol/min/mg prot) (D). Activities were measured in microsomes or cytosol of control cells (CTR) and 24 h treated cells. Results are expressed as percentages of control values. Mean ± SE of cells from five rats. • Significantly different from controls, p<0,05. ••p<0.01. ••• p<0.001. ○○○ Significantly different from H₂O₂, p<0.001.</p

Primer pairs, annealing temperature and product size for RT-PCR experiments.

<p>Primer pairs, annealing temperature and product size for RT-PCR experiments.</p

Western blot analysis heme oxygenase-1 protein.

<p>In microsomes (50 µg) of control (CTR) cells and cells treated for 24 h with Lisosan G or Lisosan G+H₂O₂. Microsomal samples were subjected to SDS-PAGE, electrophoretically transferred to a nitrocellulose membrane, and probed with polyclonal antibodies raised against rat HO-1. Densitometric analysis of the western blot data are shown in the histogram. The results have been normalized to β-actin levels and are expressed as percentages of control. Mean ± SE of cells from five independent experiments using five rats. ••• Significantly different from controls, p<0.001. ○○○ Significantly different from H₂O₂, p<0.001.</p

Effect of Lisosan G on GSH levels.

<p>The activity was measured in homogenates of 24 h treated hepatocytes. Results are expressed as percentages of control activity (125.67±28.04 nmol/mg prot.). Mean ± SE of cells from five rats. ••Significantly different from control, p<0.01. ••• p<0.001. ○○○ Significantly different from H₂O₂, p<0.001.</p

A representative RT-PCR analysis.

<p>NQO1 (A), HO-1 (B) genes performed with 25, 27 and 29 cycles in primary rat hepatocytes of control (CTR) and treated with Lisosan G or Lisosan G+H₂O₂. PCR products were separated by electrophoresis on agarose gels and stained with ethidium bromide. Quantitative representation of the RT-PCR analysis is reported in the histograms and the results have been normalized to β-actin levels and are expressed as percentages of control. Mean ± SE of cells from five independent experiments using five rats. •• Significantly different from controls, p<0.01. ••• p<0.001. ○ Significantly different from H₂O₂, p<0.05. ○○○ p<0.001.</p

Western Blotting analysis of NF-kB.

<p>In nuclear extracts of control cells (CTR) and cells treated with Lisosan G or Lisosan G+H₂O₂. Protein samples (30 µg) were subjected to SDS-PAGE, electrophoretically transferred to a nitrocellulose membrane, and probed with polyclonal antibodies raised against rat NF-kB. Densitometric analysis of the western blot data are shown in the histogram. The results have been normalized to β-actin levels and are expressed as percentages of control. Mean ± SE of cells from five independent experiments using five rats. ••• Significantly different from controls, p<0.001. ○○○ Significantly different from H₂O₂, p<0.001.</p

DataSheet_1_Characterization of the endophytic bacterial community of Bituminaria bituminosa plant grown in vitro and its interaction with the plant extract.pdf

IntroductionBituminaria bituminosa is a medicinal plant recognized for its phytochemicals, such as furanocoumarins, pterocarpans, and flavonoids. Since the secondary metabolism is influenced by the plant-endophyte interactions, the endophytic bacterial community of B. bituminosa was explored and the possible interactions with the plant were described.Materials and methodsDifferent bacterial strains were isolated from different organs of in vitro plants as shoots, roots, and seeds. The bacterial strains were identified and phenotypically characterized for different traits; strains were also exposed to different concentrations of B. bituminosa plant extract showing different susceptibility, probably determined by different secondary metabolites produced by the plant in the different organs (i.e. aerial parts and roots).Results and discussionBacterial strains showed different phenotypic characteristics; the 6 detected haplotypes were dominated by a single species related to Stenotrophomonas rhizophila. Endophytes isolated from the aerial parts produced a higher indole-3-acetic acid (IAA) amount than those of the roots, while all strains were unable to produce biosurfactants and antagonistic activity toward the other strains. The research opens new perspectives for future analysis addressed to test the susceptibility of the endophytic bacterial community of B. bituminosa toward the pure compounds extracted from the plants, and to investigate the role of these compounds on the distribution of endophytes within the different plant tissues.</p

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

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

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