13 research outputs found
135-Day Interventions of Yam Dioscorin and the Dipeptide Asn-Trp (NW) To Reduce Weight Gains and Improve Impaired Glucose Tolerances in High-Fat Diet-Induced C57BL/6 Mice
The
C57BL/6J mice were fed a 135-day normal diet or a high-fat
diet (HFD) without, or concurrent with, a single yam dioscorin (80
mg/kg) or dipeptide NW (40 mg/kg) intervention every day. The final
body weights (g) of mice were 26.1 ± 1.4, 34.97 ± 2.1, 31.75
± 2.6, and 31.66 ± 3.1, respectively, for normal diet-fed,
HFD-fed, dioscorin-intervened, and NW-intervened group. The mice in
both intervened groups showed similar less weight gains and had significant
differences (<i>P</i> < 0.05) compared to those in the
HFD group under the same cumulative HFD intakes. The blood biochemical
index of mice with dioscorin interventions showed significantly lower
contents in total cholesterol and low-density lipoprotein, and NW
interventions showed significantly lower total triglyceride contents
compared to those of the HFD group (<i>P</i> < 0.05).
Both intervened mice exhibited similar reductions in total visceral
lipid contents and have significant differences compared to those
of the HFD group (<i>P</i> < 0.05). The dioscorin intervention
was better than NW interventions in lowering blood glucose levels
by oral glucose tolerance tests and both showed significant differences
(<i>P</i> < 0.05) compared to those in the HFD group.
Yam dioscorin or dipeptide NW will potentially be used for preventive
functional foods of less body weight gains and impaired glucose tolerance
controls, which require further clinical trial investigations
<i>N</i>‑Hydroxycinnamide Derivatives of Osthole Presenting Genotoxicity and Cytotoxicity against Human Colon Adenocarcinoma Cells in Vitro and in Vivo
Osthole
is extracted from the Chinese herbs <i>Cnidium monnieri</i> and <i>Angelica pubescens</i>, and it was found to have
antitumor activity in vitro and in vivo. A series of osthole derivatives
have been synthesized, and the <i>N</i>-hydroxycinnamide
derivatives of osthole, WJ1376-1 and WJ1398-1 were found to have the
greatest potential against human colon adenocarcinoma cells. In contrast
to the parental osthole, both WJ1376-1 and WJ1398-1 were found to
induce multinucleation and polyploidy by microscopic observation and
flow cytometry. WJ1376-1 and WJ1398-1 significantly activated ataxia
telangiectasia and rad3 related (ATR) kinase, which triggered activation
of the checkpoint kinase 2 (Chk2) signaling pathway and then down
regulated Cdc25 phosphatase and Cdc2/cyclin B kinase activities. WJ1376-1
and WJ1398-1 also inhibited the phosphorylation of Aurora A kinase,
which is associated with important processes during mitosis. The presence
of a “comet” DNA fragment and phosphorylation of p53
at Ser 15 clearly indicated that DNA damage occurred with WJ1376-1
and WJ1398-1 treatment. WJ1376-1 and WJ1398-1 ultimately induced apoptosis
as evidenced by the upregulation of Bad and activation of caspases-3,
-7, and -9. Furthermore, WJ1376-1 and WJ1398-1 also showed a great
effect in attenuating tumor growth without affecting the body weight
of xenograft nude mice. Taken together, these results suggest that
the toxic activities of WJ1376-1 and WJ1398-1 were dissimilar to that
of the parental osthole, which can induce cell polyploidy and G<sub>2</sub>/M cell cycle arrest in colon adenocarcinoma cells and may
provide a potential therapeutic target for colon cancer treatment
in the future
Modulation the alternative splicing of <i>GLA</i> (IVS4+919G>A) in Fabry disease
<div><p>While a base substitution in intron 4 of <i>GLA</i> (IVS4+919G>A) that causes aberrant alternative splicing resulting in Fabry disease has been reported, its molecular mechanism remains unclear. Here we reported that upon IVS4+919G>A transversion, H3K36me3 was enriched across the alternatively spliced region. PSIP1, an adapter of H3K36me3, together with Hsp70 and NONO were recruited and formed a complex with SF2/ASF and SRp20, which further promoted <i>GLA</i> splicing. Amiloride, a splicing regulator in cancer cells, could reverse aberrant histone modification patterns and disrupt the association of splicing complex with <i>GLA</i>. It could also reverse aberrant <i>GLA</i> splicing in a PP1-dependant manner. Our findings revealed the alternative splicing mechanism of <i>GLA</i> (IVS4+919G>A), and a potential treatment for this specific genetic type of Fabry disease by amiloride in the future.</p></div
A hypothetical model for the alternative splicing of <i>GLA</i> (IVS4 + 919G>A).
<p>A hypothetical model for the alternative splicing of <i>GLA</i> (IVS4 + 919G>A).</p
Alterations in histone modification patterns after amiloride treatment.
<p>ChIP assays were performed with antibodies to the indicated histone modifications on the cryptic exon area in Int4 of <i>GLA</i> in normal cells or in FD cells treated with or without amiloride. Results were expressed as a fraction of histone H3 after normalization to input values and presented as a mean values ± standard deviation from three independent experiments. Asterisk represents significant difference (<i>p</i>-value < 0.05). FD cells, Fabry disease cells; Amil, amiloride.</p
Effects of RNA-associated proteins on the cryptic exon of <i>GLA</i> transcripts.
<p>(A-C) Virus-mediated shRNA knockdown of various genes as indicated. SMYD2 and NSD1 are histone methyltransferases which preferentially methylates Lys-36 of histone H3. (D) RNA-chromatin immunoprecipitations (RNA-ChIP) analysis on the cryptic exon area in Int4 of <i>GLA</i> was performed using antibodies against SF2/ASF and SRp20 with IgG as a control. (E) Co-immunoprecipitation results using the indicated antibodies for immunoprecipitation and analyzing by western blotting. FD cells, Fabry disease cells; NC: negative control; WB, western blot; SMYD2, SET and MYND domain containing 2; NSD1, nuclear receptor binding SET domain protein 1.</p
Effects of amiloride on the regulation of <i>GLA</i> (IVS4 + 919G>A) splicing.
<p>Cells were treated with different concentrations of amiloride for 24 hours and then harvested for RT-PCR analysis (A), or Western blot analysis (B,D). Actin and Histone H3 were used as internal standards. (C) The result of enzyme activity assay from FD cells after the treatment with or without amiloride for 24 hours. Data were presented as the mean ± standard deviation from three independent experiments. Asterisk represents significant difference (<i>p</i>-value < 0.05). (E) RT-PCR and Western blot results from cells pretreated with (+) or without (-) okadaic acid and then exposed to amiloride for 24 hours. FD cells, Fabry disease cells; Amil, amiloride; OA, okadaic acid.</p
Schematic representation of <i>GLA</i> transcripts.
<p>(A) Schematic illustration of <i>GLA</i> with positions of the biotin-labeled RNA probes for pull-down assays. (B) Putative regulatory motifs and binding sites for the splicing factors as determined by ESEfinder<sup>a</sup>, Spliceaid2<sup>b</sup>, Human Splicing Finder <sup>c</sup>, and our pull-down experiments*. ESS motifs and hnRNPA1 binding motifs were predicted to be disturbed upon IVS4 + 919G>A transversion. (C) Analyses of the RNA folding of <i>GLA</i> (IVS4 + 919G/A). The alternatively spliced 57 nucleotide sequence is highlighted in gray and the boxes indicate the alterations in RNA folding.</p
Effects of proteins associated with the cryptic exon area in Int4 of <i>GLA</i>.
<p>(A) Schematic illustration of <i>GLA</i> with positions of the biotin-labelled DNA probes for pull-down assays. (B) ChIP analysis on the cryptic exon area in Int4 of <i>GLA</i> was performed using antibodies against HSP70, NONO, and H3K36me3 with IgG as a control. (C) Co-immunoprecipitation results using anti- HSP70 or anti-NONO antibody for immunoprecipitation and analyzing by western blotting. Nonimmune IgG was used as negative control. (D) Fabry disease cells were infected with lentiviruses expressing shRNAs targeting <i>HSP70</i> or <i>NONO</i>, or treated with 4β-hydroxywithanolide E (4HWE). Messenger RNA was extracted after 48 hours infection or 24 hours 4HWE treatment followed by RT-PCR analysis. WB, western blot; NC: negative control.</p