6 research outputs found
Effect of BEX on lipotoxic MCP-1 production.
<p>Production of MCP-1 was measured in the cell media supernatant (AāC) and cytosolic fraction (DāF) of 3T3-L1 (A, D), C2C12 (B, E), and Hepa6 (C, F) murine cells treated with palmitic acid (PA, 0.4 mM) in combination with BEX (125 Āµg/ml or 0.5%, v/v) or ethanol vehicle (0.5%, v/v). Concentrations were determined by cytometric bead array immunodetection against a reconstituted MCP-1 standard. Mean Ā± standard error over 3 trials, n ā„300 beads in any trial. Mean results were compared between treatments of the same cell type and at the same time point. Differences between means are statistically significant if columns do not share any common letters (p<0.05, one-way ANOVA with Bonferroniās post hoc test).</p
Effect of BEX on MCP-1 mRNA expression.
<p>Levels of MCP-1 mRNA expression in 3T3-L1 (A), C2C12 (B), and Hepa6 (C) murine cells in response to palmitic acid (PA, 0.4 mM) in combination with BEX (125 Āµg/ml or 0.5%, v/v) or ethanol vehicle (0.5%, v/v) at different time points and differentiation states (3T3-L1 and C2C12). Fold change calculated via comparative cycle threshold (āĪĪCt) normalized to beta-glucuronidase (GUSĪ²). Other housekeeping genes such as 18 S ribosomal RNA (18 S), hypoxanthine-guanine phosphoribosyltransferase (HPRT), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) were also used and yielded similar results as GUSĪ². Mean Ā± standard deviation of at least 4 qRT-PCRs per cell type. Mean results were compared between treatments of the same cell type and at the same time point. Differences between means are statistically significant if columns do not share any common letters (p<0.05, one-way ANOVA with Tukeyās multiple comparison test).</p
Molecular structures of tricin and 7-O-methyl-tricin (7 MT), two compounds isolated from BEX.
<p>Molecular structures of tricin and 7-O-methyl-tricin (7 MT), two compounds isolated from BEX.</p
Dose-dependent effects of tricin, 7-O-methyl-tricin (7 MT), Fraction K, and BEX on lipotoxic MCP-1 production and cell viability.
<p>(A) MCP-1 concentrations in cell culture media from 3T3-L1 cells cultured in 96 well plates and treated for 20 hours with PA (0.4 mM) and one of the following: tricin (black triangles), 7 MT (light grey hollow squares), Fraction K (grey diamonds), or BEX (dark grey crosses). Concentrations were determined by cytometric bead array immunodetection against a reconstituted MCP-1 standard. Mean Ā± standard error of 3 samples per dose, n ā„300 beads per sample. (B) Viability of 3T3-L1 cells treated with tricin (black triangles), 7 MT (light grey hollow squares), Fraction K (grey diamonds), or BEX (dark grey crosses) added to maintenance medium. All values are normalized to 3T3-L1 cells from the same plate treated with maintenance medium without flavonoids or PA (normalized average represented by a red dashed line). Mean Ā± standard error of three trials, n ā„3 per trial. (C) Viability of 3T3-L1 cells treated with ethanol vehicle (outlined white), Fraction K (grey), or BEX (black). Mean Ā± standard error of three trials, n ā„4 per trial. All values normalized to 3T3-L1 cells from the same plate treated with only maintenance medium (normalized average represented by a red dashed line). <i>p</i> values were obtained by performing a Studentās <i>t</i>-test between doses.</p
Protective effect of BEX on palmitic acid (PA)-induced lipotoxicity.
<p>3T3-L1 (A), and Hepa6 cells (B) were grown to confluency in 96-well plates and treated with 0.4 mM PA for 24 hours in combination with BEX (125 Āµg/ml or 0.5%, v/v) or ethanol (0.5%, v/v) as a solvent control. Cell viability was measured using a MTS assay. Mean Ā± standard deviation over 5 trials, n ā„3 wells per trial. Differences between means are statistically significant if the columns do not share any common letters (p<0.001, one-way ANOVA, Tukeyās multiple comparison test).</p
Global and Targeted Metabolomics Evidence of the Protective Effect of Chinese Patent Medicine <i>Jinkui Shenqi</i> Pill on Adrenal Insufficiency after Acute Glucocorticoid Withdrawal in Rats
Glucocorticoids
are commonly used in anti-inflammatory and immunomodulatory
therapies, but glucocorticoid withdrawal can result in life-threatening
risk of adrenal insufficiency. Chinese patented pharmaceutical product <i>Jinkui Shenqi</i> pill (JKSQ) has potent efficacy on clinical
adrenal insufficiency resulting from glucocorticoid withdrawal. However,
the underlying molecular mechanism remains unclear. We used an animal
model to study JKSQ-induced metabolic changes under adrenal insufficiency
and healthy conditions. SpragueāDawley rats were treated with
hydrocortisone for 7 days with or without 15 days of JKSQ pretreatment.
Sera were collected after 72 h hydrocortisone withdrawal and used
for global and free fatty acids (FFAs)-targeted metabolomics analyses
using gas chromatography/time-of-flight mass spectrometry and ultraperformance
liquid chromatography/quadruple time-of-flight mass spectrometry.
Rats without hydrocortisone treatment were used as controls. JKSQ
pretreatment normalized the significant changes of 13 serum metabolites
in hydrocortisone-withdrawal rats, involving carbohydrates, lipids,
and amino acids. The most prominent effect of JKSQ was on the changes
of FFAs and some [product FFA]/[precursor FFA] ratios, which represent
estimated desaturase and elongase activities. The opposite metabolic
responses of JKSQ in adrenal insufficiency rats and normal rats highlighted
the ā<i>Bian Zheng Lun Zhi</i>ā (treatment
based on ZHENG differentiation) guideline of TCM and suggested that
altered fatty acid metabolism was associated with adrenal insufficiency
after glucocorticoid withdrawal and the protective effects of JKSQ