<i>Syzygium aromaticum</i> (L.) elicits lifespan extension and attenuates age-related Aβ-induced proteotoxicity in <i>Caenorhabditis elegans</i>

<p>Clove (<i>Syzygium aromaticum</i>) is a popular medicinal plant which has been traditionally used in India as spice and medicine to counter various ailments. <i>However, the stress modulatory and antiaging potential of this plant is yet to be characterized.</i> Therefore, the present study evaluates the effect of clove oil (CO) on oxidative stress, lifespan, mobility, and the expression of aging-related proteins using <i>Caenorhabditis elegans</i> model system. The CO (10 ppm) was found to extend mean lifespan in worms by 21.4% (<i>p</i> < 0.001) under normal and by 63% (<i>p</i> < 0.0001) under juglone-induced oxidative stress conditions. The extension of mean lifespan in <i>mev</i>-<i>1</i> mutant and elevated expression of <i>gst</i>-4 and <i>sod</i>-3 confirmed stress modulatory effects of CO. Additionally, the CO reduced intracellular ROS and Aβ_1–42 proteotoxicity. Altogether, the present study unravels the anti-aging and stress modulatory potential of CO and suggests CO as a potential pharmaceutical entity in modulating aging process.</p

Effect of fC-MSC on LV functions: Bar diagrams showing ejection fraction, end systolic volume, end diastolic volume, and left ventricular myo-mass, measured at 1 week after MI (before fC-MSC therapy) and 4 weeks after fC-MSC therapy using gated SPECT analysis.

<p>Values shown are mean ± SEM (n = 6); *P<0.05, *P<0.01, *P<0.001 saline group after cell therapy vs before cell therapy (within the group);^#P<0.05,^#P<0.01, ^#P<0.001 fC-MSC group after cell therapy vs before cell therapy (within the group);^†P<0.05, ^†P<0.01, ^†P<0.001 fC-MSC group (after cell therapy) vs saline group (after cell therapy).</p

Effect of fC-MSC on gene expression of growth factors in rats with acute myocardial injury.

<p>Bar diagrams showing fold change expression of VEGF, b-FGF, IGF-1, and HGF in saline treated and fC-MSC treated ischemic hearts 4 weeks after fC-MSC administration. Values shown are mean ± SEM of 6 experiments, *P<0.05, **P<0.01, ***P<0.001: fC-MSC vs saline treatment.</p

fC-MSC inhibit the apoptosis in rats with acute myocardial injury.

<p>(a) Representative immunofluorescence photomicrographs of saline and fC-MSC treated hearts 4 weeks after fC-MSC therapy showing (a-i & b-i) Overlay; (a-ii & b-ii) Tunel positive cells (green dye) counter stain with (a-iii & b-iii) Hoechst dye respectively (B) TUNEL apoptotic index showing significant decrease in apoptotic cells in fC-MSC treated compared to saline treated hearts. Values shown are mean ± SEM (n = 6). **P<0.01 fC-MSC treated vs saline treated hearts. (C) Representative immune-blots showing expression of BAX and BCL2 in saline and fC-MSC treated rats and (D) their relative density. Densitometric analysis was applied for comparison of relative protein expression. Values expressed Mean ± SE (n = 6), *P<0.05, **P<0.01, ***P<0.001: fC-MSC treated vs. saline treated group.</p

Gated SPECT analysis of fC-MSC therapy in rats with MI.

<p>***P<0.001,</p><p>**P<0.01 = Healthy Control Vs MI Baseline;</p>††<p>P<0.01 = MI Baseline Vs MI+Saline;</p>‡‡‡<p>P<0.001 ^‡‡P<0.01,</p>‡<p>P<0.05 = MI baseline Vs MI+fC-MSC;</p>§§§<p>P<0.001,</p>§§<p>P<0.01 = MI+Saline Vs MI+fC-MSC.</p

fC-MSC attenuate the myocardial fibrosis in rats with acute myocardial injury (A) Representative heart sections stained with Massons Trichome showing decrease in myocardial fibrosis in fC-MSC treated compared to saline treated hearts 4 weeks after fC-MSC administration.

<p>(B) Bar diagram showing percentage of fibrosis in saline treated and fC-MSC treated hearts. Values shown are mean ± SEM (n = 6). **P<0.05, **P<0.01, ***P<0.001: fC-MSC treated vs saline treated hearts.</p

Morphology and characterization of fC-MSC (A) Representative photomicrograph (10X, 20 µm) of fC-MSC in culture showing spindle shaped morphology.

<p>(B) Representative flow cytometric dot-plots showing that fC-MSC are (a) CD29+/CD45−; (b) CD44+/CD45−; (c) CD73+/CD31−; (d) CD90+/HLA-DR−; (e) CD105+/HLA-DR−. (C) Representative photomicrographs (10X, 20 µm) showing differentiation of fC-MSC into Osteoblasts (a-i: differentiated cells positive for Alizarin red stain, and a-ii: control cells negative for Alizarin red stain) and Adipocytes (b-i: differentiated cells positive for oil red O stain, and b-ii: control cells negative for oil red O stain).</p

Effect of fC-MSC on LV perfusion: Representative SPECT perfusion images and polar-maps obtained at 1 week after MI (before fC-MSC therapy) and 4 weeks after fC-MSC therapy.

<p>(A) Serial 99mTc-sestamibi perfusion images obtained in SPECT short axis (SA), horizontal long axis (HLA), vertical long axis (VLA) of MI hearts treated with saline and fC-MSC (B) Corresponding polar-maps. The perfusion images (A) and the polar-maps (B) show a myocardial-flow defect in the anterolateral wall of left ventricle in both the groups. However, the hearts treated with fC-MSC demonstrate a smaller ischemic lesion (region of deficit) and a better perfusion in the MI segments.</p

Effect of fC-MSC on LV functions: Bar diagrams showing ejection fraction, end systolic volume, end diastolic volume, and left ventricular myo-mass, measured at 1 week after MI (before fC-MSC therapy) and 4 weeks after fC-MSC therapy using gated SPECT analysis.

<p>Values shown are mean ± SEM (n = 6); *P<0.05, *P<0.01, *P<0.001 saline group after cell therapy vs before cell therapy (within the group);^#P<0.05,^#P<0.01, ^#P<0.001 fC-MSC group after cell therapy vs before cell therapy (within the group);^†P<0.05, ^†P<0.01, ^†P<0.001 fC-MSC group (after cell therapy) vs saline group (after cell therapy).</p

Primers used for the Real Time PCR study.

<p>Primers used for the Real Time PCR study.</p