Preventive effects of TCM on TBARS formation in tissues under whiskers cut stress in mice

<p><b>Copyright information:</b></p><p>Taken from "Psychological Stress-Induced Oxidative Stress as a Model of Sub-Healthy Condition and the Effect of TCM"</p><p></p><p>Evidence-based Complementary and Alternative Medicine 2006;4(2):195-202.</p><p>Published online 2 Nov 2006</p><p>PMCID:PMC1876613.</p><p>© 2006 The Author(s).</p> SMS or LGZGT were the same as in . At 12 h after whiskers removal, tissues were removed to determine TBARS. Data represent means ± SD ( = 6 mice). < 0.05 or < 0.01 versus the untreated control mice. < 0.05 or < 0.01 versus stressed mice

Effect of whiskers cut stress and Shengmai San on locomotive activities of mice at night

<p><b>Copyright information:</b></p><p>Taken from "Psychological Stress-Induced Oxidative Stress as a Model of Sub-Healthy Condition and the Effect of TCM"</p><p></p><p>Evidence-based Complementary and Alternative Medicine 2006;4(2):195-202.</p><p>Published online 2 Nov 2006</p><p>PMCID:PMC1876613.</p><p>© 2006 The Author(s).</p> SMS was orally administered to mice once a day for 5 days, after which the mice were distressed by whiskers removal. Three types of behavioral movements (small and large movements, and rearing) were then recorded in five mice for 12 h at night. Values represent the means ± SD (frequency within 12 h per five mice) of three independent experiments.  = 5. < 0.05, < 0.01 compared to the control (unstressed) mice; < 0.05, < 0.01 compared to the stressed mice

Sch B suppressed the cardiac morphological changes and DNA damage.

<p>Typical photomicrographs of heart tissue from group N (A), group V (B) and group Sch B (C–E). Cytoplasmic vacuolation and myofibrillar disorganization in the doxorubicin group were effectively attenuated Sch B treatment (haematoxylin and eosin ×200). (F–J) Representative photomicrographs of DNA damage 5 days after Dox injection. DNA damage detected by single cell gel electrophoresis assay (Comet assay) in cardiomyoctes of all groups at 200x magnification. (K–L) Quantification of cardiomyocyte DNA damage analyzed by digital imaging casp-software (<a href="http://casp.sourceforge.net/" target="_blank">http://casp.sourceforge.net/</a>). **<i>P <</i> 0.01 vs. group N mice; ^#<i>P <</i> 0.05 and ^##<i>P <</i> 0.01 vs. group V mice.</p

Sch B attenuates Dox-induced myocardial ROS and oxidative stress, and enhance antioxidant enzyme.

<p>(A) MDA level, (B) Nitrite level, (C) Glutathione level and (D) Superoxide production by LV homogenates. **<i>P <</i> 0.01 vs. group N mice; ^#<i>P <</i> 0.05 and ^##<i>P <</i> 0.01 vs. group V mice.</p

Sch B inhibits Dox induced NADPH oxidase subunits.

<p>(A–D) Representative Western immunoblots and densitometry analysis using Scion image software for p47phox, p67phox and gp91phox in group N, group V and Sch B treated mice; Blots were normalized against GAPDH. Each bar represents means ± S.E. (n = 3–5). **<i>P <</i> 0.01 vs. group N mice; ^#<i>P <</i> 0.05 and ^##<i>P <</i> 0.01 vs. group V mice.</p

Sch B attenuates Dox-induced hydroxyl radical.

<p>(A, B) Representative ESR spectra and analysis of the hydroxyl radical signal relative to the internal standard of manganese ion. Hydroxyl radical signals were not detected (ND) in hearts of group N and group Sch B-100. Mn (3) and Mn (4) indicate the internal standard signals of manganese ion (Mn²⁺).</p

Squalene enhances γIR toxicity.

<p>Relative cell viability was measured by a clonogenic assay. (A) Cytotoxicity of squalene (0, 1, 3, 10, 30, and 100 μM) was measured after 14 days of incubation. (B) and (C) Impact of squalene on radiation toxicity. The cells were subjected to 1 h of pre-incubation and 14 days of post-incubation with squalene (0, 30, and 100 μM) after UVC irradiation (0, 25, 50, and 100 J/m²) or γIR (0, 2, and 6 Gy). Data are presented as the mean ± S.D. of three independent experiments (*P < 0.05 vs. untreated control).</p

Squalene uptake in cultured cells.

<p>Squalene uptake by A549 adenocarcinoma cells. The cells were incubated with various concentrations of squalene (0–300 μM) for 3 h, washed in PBS, and harvested. Squalene was extracted from the cells using chloroform/methanol, and the amount of squalene was determined by GC-MS.</p

Sch B inhibits cardiac apoptosis, caspase-3 positive cells and superoxide production.

<p>(A–E) Myocardial tissue sections TUNEL stained for apoptotic nuclei in group N (A), group V (B) and Sch B treated mice (C–E) at 200x magnification. (F–J) Representative photomicrographs of LV sections showing caspase-3 immunofluoroscence identified by immunohistochemical staining with anti-capsase-3 antibody, group N (F), group V (G) and Sch B treated mice (H–J) (400x); Intense immunofluoroscence was detected in LV sections of group V mice. (K–O) In situ superoxide production (bright area) using dihydroethidium (DHE) staining in hearts of group N (K), group V (L) and Sch B treated mice (M–O) at 200x magnification.</p

Squalene does not inhibit ATM and ATR kinases.

<p>ATM (A) and ATR (B) kinase activities were measured <i>in vitro</i> using PHAS-I as the substrate in the presence of squalene (1, 3, 10, 30, 100, and 300 μM). HEK293T cells were transfected with flag-tagged ATM- or ATR-wt plasmid. ATM and ATR protein kinases were purified by immunoprecipitation using anti-Flag^® M2 antibody and protein-G sepharose. Kinase activity was monitored for 20 min at 30°C. Data are presented as the mean ± S.D. of three independent experiments. Caffeine (2 and 10 mM) was used as an ATM/ATR inhibitor.</p