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<input type="email" aria-label="Recipient Email Address" placeholder="email@example.com" id="email-to" class="email-to" pattern="^[a-zA-Z0-9_.+-]+@[a-zA-Z0-9-]+\.[a-zA-Z0-9-.]+$" maxlength="100" required>
</div>
<div class="action-panel-control-wrap">
<label for="email-from" class="action-panel-label">
From:
</label>
<input type="email" aria-label="Sender Email Address" placeholder="email@example.com" id="email-from" class="email-from" pattern="^[a-zA-Z0-9_.+-]+@[a-zA-Z0-9-]+\.[a-zA-Z0-9-.]+$" maxlength="256">
</div>
<div class="action-panel-control-wrap">
<label for="email-citation-format" class="action-panel-label">
Format:
</label>
<select id="email-citation-format" name="citation-format" class="action-panel-selector email-citation-format">
<option selected="selected" value="summary">Summary</option>
<option value="summary-text">Summary (text)</option>
<option value="abstract">Abstract</option>
<option value="abstract-text">Abstract (text)</option>
</select>
</div>
<div class="include-supplemental-container">
<input type="checkbox" aria-label="Include MeSH and other data" name="include-supplemental" id="email-include-supplemental" class="email-include-supplemental">
<label for="email-include-supplemental" class="email-include-supplemental-label">MeSH and other data</label>
</div>
<div class="form-field recaptcha ">
<div class="g-recaptcha" id="id-recaptcha" data-sitekey="6LfsWHMdAAAAAClKbtOpjQ2pMjgsGxvv7NdZW9uI"></div>
</div>
<div id="captcha-error-message" class="usa-input-error-message captcha-validation-message" role="alert"></div>
<div class="action-panel-actions">
<button class="action-panel-submit"
type="submit"
data-loading-label="Sending..."
data-ga-category="save_share"
data-ga-action="email"
data-ga-label="send">
Send email
</button>
<button class="action-panel-cancel"
aria-label="Close 'Email citations' panel"
ref="linksrc=close_email_panel"
aria-controls="email-action-panel"
aria-expanded="false"
data-ga-category="save_share"
data-ga-action="email"
data-ga-label="cancel">
Cancel
</button>
</div>
<input type="hidden" name="email-search-details" value="" />
<input type="hidden" name="email-search-details-hash" value="0e42663a6c3bd85498fcb88798998fed7bfdc45d457db35281e41afe13cc0524" />
</form>
</div>
</div>
<div id="collections-action-panel"
class="collections-action-panel action-panel in-progress-dots-panel"
aria-hidden="true"
data-collections-open-panel-enabled="false"
data-collections-open-panel-url-hash="#open-collections-panel">
<div class="inner-wrap">
<h3 class="action-panel-heading">
Add to Collections
</h3>
<form id="collections-action-panel-form"
class="collections-action-panel-form action-panel-content action-form action-panel-smaller-selectors"
data-existing-collections-url="/list-existing-collections/"
data-add-to-existing-collection-url="/add-to-existing-collection/"
data-create-and-add-to-new-collection-url="/create-and-add-to-new-collection/"
data-get-article-ids-by-search-url="/get-article-ids-by-search/"
data-myncbi-max-collection-name-length="100"
data-add-to-collection-max-amount="1000"
data-collections-root-url="https://www.ncbi.nlm.nih.gov/myncbi/collections/">
<input type="hidden" name="csrfmiddlewaretoken" value="tjvTMg4THnEvebrvKI4ncRNOvsmbwWd9K6fYp6xfhYrZ8Cxln1HpGZjs3w4npxLt">
<div class="choice-group" role="radiogroup">
<ul class="radio-group-items">
<li>
<input type="radio"
id="collections-action-panel-new"
class="collections-new"
name="collections"
value="new"
data-ga-category="save_share"
data-ga-action="collections"
data-ga-label="collections_radio_new">
<label for="collections-action-panel-new">Create a new collection</label>
</li>
<li>
<input type="radio"
id="collections-action-panel-existing"
class="collections-existing"
name="collections"
value="existing"
checked="true"
data-ga-category="save_share"
data-ga-action="collections"
data-ga-label="collections_radio_existing">
<label for="collections-action-panel-existing">Add to an existing collection</label>
</li>
</ul>
</div>
<div class="controls-wrapper">
<div class="action-panel-control-wrap new-collections-controls">
<label for="collections-action-panel-add-to-new" class="action-panel-label required-field-asterisk">
Name your collection:
</label>
<input
type="text"
name="add-to-new-collection"
id="collections-action-panel-add-to-new"
class="collections-action-add-to-new"
pattern="[^&quot;&amp;=&lt;&gt;\/]*" title="The following characters are not allowed in the Name field: &quot;&amp;=&lt;&gt;/"
maxlength="100"
data-ga-category="save_share"
data-ga-action="create_collection"
data-ga-label="non_favorties_collection">
<div class="collections-new-name-too-long usa-input-error-message selection-validation-message">
Name must be less than 100 characters
</div>
</div>
<div class="action-panel-control-wrap existing-collections-controls">
<label for="collections-action-panel-add-to-existing" class="action-panel-label">
Choose a collection:
</label>
<select id="collections-action-panel-add-to-existing"
class="action-panel-selector collections-action-add-to-existing"
data-ga-category="save_share"
data-ga-action="select_collection"
data-ga-label="($('#collections-action-add-to-existing').val() === 'Favorites') ? 'Favorites' : 'non_favorites_collection'">
</select>
<div class="collections-retry-load-on-error usa-input-error-message selection-validation-message">
Unable to load your collection due to an error<br>
<a href="#">Please try again</a>
</div>
</div>
</div>
<div class="action-panel-actions">
<button class="action-panel-submit"
type="submit"
data-loading-label="Adding..."
data-pinger-ignore
data-ga-category="save_share"
data-ga-action="collections"
data-ga-label="add">
Add
</button>
<button class="action-panel-cancel"
aria-label="Close 'Add to Collections' panel"
ref="linksrc=close_collections_panel"
aria-controls="collections-action-panel"
aria-expanded="false"
data-ga-category="save_share"
data-ga-action="collections"
data-ga-label="cancel">
Cancel
</button>
</div>
</form>
</div>
</div>
<div id="bibliography-action-panel"
class="bibliography-action-panel action-panel in-progress-dots-panel"
aria-hidden="true"
data-bibliography-open-panel-enabled="false"
data-bibliography-open-panel-url-hash="#open-bibliography-panel">
<div class="inner-wrap">
<h3 class="action-panel-heading">
Add to My Bibliography
</h3>
<form id="bibliography-action-panel-form"
class="bibliography-action-panel-form action-panel-content action-form action-panel-smaller-selectors"
data-add-to-bibliography-max-amount="100"
data-add-to-bibliography-batch-size="10"
data-bibliography-delegates-url="/list-bibliography-delegates/"
data-add-to-bibliography-url="/add-to-bibliography/"
data-get-article-ids-by-search-url="/get-article-ids-by-search/"
data-mybib-root-url="https://www.ncbi.nlm.nih.gov/myncbi/collections/mybibliography/">
<input type="hidden" name="csrfmiddlewaretoken" value="tjvTMg4THnEvebrvKI4ncRNOvsmbwWd9K6fYp6xfhYrZ8Cxln1HpGZjs3w4npxLt">
<div class="action-panel-control-wrap bibliographies-controls">
<div class="choice-group">
<ul class="bibliographies-action-add radio-group-items">
<li>
<input name="bibliography" id="my-bibliography" class="my-bibliography" type="radio" checked/>
<label for="my-bibliography">My Bibliography</label>
</li>
</ul>
</div>
</div>
<div class="bibliographies-retry-load-on-error usa-input-error-message selection-validation-message">
Unable to load your delegates due to an error<br>
<a href="#">Please try again</a>
</div>
<div class="action-panel-actions">
<button class="action-panel-submit"
type="submit"
data-loading-label="Adding..."
data-pinger-ignore>
Add
</button>
<button class="action-panel-cancel"
aria-label="Close 'Add to bibliography' panel"
ref="linksrc=close_bibliography_panel"
aria-controls="bibliography-action-panel"
aria-expanded="false"
data-ga-category="save_share"
data-ga-action="mybib"
data-ga-label="cancel">
Cancel
</button>
</div>
</form>
</div>
</div>
<div id="saved-search-action-panel" class="saved-search-action-panel action-panel " aria-hidden="true"
data-saved-search-open-panel-enabled="false"
data-saved-search-open-panel-url-hash="#open-saved-search-panel">
<div class="inner-wrap">
<h2 class="action-panel-heading">
Your saved search
</h2>
<form id="saved-search-action-panel-form"
class="saved-search-action-panel-form action-panel-content action-form"
data-create-saved-search-url="/create-saved-search/"
data-try-search-terms-url="/try-search-term/"
data-saved-search-root-url="https://www.ncbi.nlm.nih.gov/myncbi/searches/">
<input type="hidden" name="csrfmiddlewaretoken" value="tjvTMg4THnEvebrvKI4ncRNOvsmbwWd9K6fYp6xfhYrZ8Cxln1HpGZjs3w4npxLt">
<div class="action-panel-control-wrap">
<label for="saved-search-name" class="action-panel-label saved-search-name-label required-field-asterisk">
Name of saved search:
</label>
<input maxlength="200"
type="text"
name="saved-search-name"
id="saved-search-name"
class="saved-search-name"
value=""
required
pattern="[^&quot;&amp;=&lt;&gt;\/]*" title="The following characters are not allowed in the Name field: &quot;&amp;=&lt;&gt;/">
</div>
<div class="action-panel-control-wrap">
<label for="saved-search-term" class="action-panel-label required-field-asterisk">
Search terms:
</label>
<textarea name="saved-search-term" id="saved-search-term" class="saved-search-term" required></textarea>
</div>
<div class="test-search-term-wrap">
<a href="#" class="try-search-term">Test search terms</a>
</div>
<div class="choice-group action-panel-extra-margin-top">
<span class="action-panel-label" id="fieldset-label">
Would you like email updates of new search results?
</span>
<fieldset id="saved-search-alert" aria-describedby="fieldset-label">
<legend class="usa-sr-only">Saved Search Alert Radio Buttons</legend>
<ul class="radio-group-items">
<li>
<input type="radio" id="saved-search-alert-yes" class="saved-search-alert-yes" name="saved-search-alert" value="yes" checked>
<label for="saved-search-alert-yes" class="action-panel-label">Yes</label>
</li>
<li>
<input aria-label="No radio input" type="radio" id="saved-search-alert-no" class="saved-search-alert-no" name="saved-search-alert" value="no">
<label for="saved-search-alert-no" class="action-panel-label">No</label>
</li>
</ul>
</fieldset>
</div>
<div class="alert-schedule-wrap">
<div class="action-panel-control-wrap">
<label class="action-panel-label">
Email:
</label>
<span aria-label="Email address" id="saved-search-email" class="action-panel-label"><span class="action-panel-label-bold"></span> (<a class="myncbi-account-settings" href="https://www.ncbi.nlm.nih.gov/account/settings/">change</a>)</span>
</div>
<div class="action-panel-control-wrap action-panel-extra-margin-top">
<label for="saved-search-frequency" class="action-panel-label">
Frequency:
</label>
<select id="saved-search-frequency" class="no-border-panel-selector saved-search-frequency">
<option value="monthly">Monthly</option>
<option value="weekly">Weekly</option>
<option value="daily">Daily</option>
</select>
</div>
<div class="action-panel-control-wrap saved-search-monthly-additional">
<label for="saved-search-monthly-on-day" class="action-panel-label">
Which day?
</label>
<select id="saved-search-monthly-on-day" class="no-border-panel-selector">
<option value="Sunday">The first Sunday</option>
<option value="Monday">The first Monday</option>
<option value="Tuesday">The first Tuesday</option>
<option value="Wednesday">The first Wednesday</option>
<option value="Thursday">The first Thursday</option>
<option value="Friday">The first Friday</option>
<option value="Saturday">The first Saturday</option>
<option value="day">The first day</option>
<option value="weekday">The first weekday</option>
</select>
</div>
<div class="action-panel-control-wrap saved-search-weekly-additional">
<label for="saved-search-weekly-on-day" class="action-panel-label">
Which day?
</label>
<select id="saved-search-weekly-on-day" class="no-border-panel-selector saved-search-weekly-on-day">
<option value="Sunday">Sunday</option>
<option value="Monday">Monday</option>
<option value="Tuesday">Tuesday</option>
<option value="Wednesday">Wednesday</option>
<option value="Thursday">Thursday</option>
<option value="Friday">Friday</option>
<option value="Saturday">Saturday</option>
</select>
</div>
<div class="action-panel-control-wrap">
<label for="saved-search-report" class="action-panel-label">
Report format:
</label>
<select id="saved-search-report" class="no-border-panel-selector saved-search-report">
<option value="DocSum">Summary</option>
<option value="DocSumText">Summary (text)</option>
<option value="Abstract">Abstract</option>
<option value="AbstractText">Abstract (text)</option>
<option value="MEDLINE">PubMed</option>
</select>
</div>
<div class="action-panel-control-wrap">
<label for="saved-search-amount" class="action-panel-label">
Send at most:
</label>
<select id="saved-search-amount" class="no-border-panel-selector saved-search-amount">
<option value="1">1 item</option>
<option value="5" selected>5 items</option>
<option value="10">10 items</option>
<option value="20">20 items</option>
<option value="50">50 items</option>
<option value="100">100 items</option>
<option value="200">200 items</option>
</select>
</div>
<div>
<input type="checkbox" id="saved-search-send-if-no-result" class="saved-search-send-if-no-result" name="saved-search-send-if-no-result">
<label for="saved-search-send-if-no-result" class="action-panel-label smaller-checkbox">
Send even when there aren't any new results
</label>
</div>
<div class="action-panel-control-wrap option-text-in-email-wrap">
<label for="saved-search-email-text" class="action-panel-label">
Optional text in email:
</label>
<textarea name="saved-search-email-text"
id="saved-search-email-text"
class="saved-search-email-text"></textarea>
</div>
</div>
<div class="action-panel-actions">
<button class="action-panel-submit"
type="submit"
data-loading-label="Saving..."
data-ga-category="save_share"
data-ga-action="alert"
data-ga-label="save">
Save
</button>
<button class="action-panel-cancel"
aria-label="Close 'Your saved search' panel"
ref="linksrc=close_saved_search_panel"
aria-controls="saved-search-action-panel"
aria-expanded="false"
data-ga-category="save_share"
data-ga-action="alert"
data-ga-label="cancel">
Cancel
</button>
</div>
</form>
</div>
</div>
<div id="citation-manager-action-panel" class="citation-manager-action-panel action-panel" aria-hidden="true">
<div class="inner-wrap">
<h2 class="action-panel-heading">
Create a file for external citation management software
</h2>
<form id="citation-manager-action-panel-form"
class="action-panel-content action-form"
action="/results-export-ids/"
data-by-search-action="/results-export-search-data/"
data-by-ids-action="/results-export-ids/"
method="post"
data-by-search-method="post"
data-by-ids-method="post">
<input type="hidden" name="csrfmiddlewaretoken" value="tjvTMg4THnEvebrvKI4ncRNOvsmbwWd9K6fYp6xfhYrZ8Cxln1HpGZjs3w4npxLt">
<input name="results-format" type="hidden" value="pubmed"/>
<div class="action-panel-actions">
<button class="action-panel-submit"
type="submit"
data-loading-label="Sending..."
data-ga-category="save_share"
data-ga-action="citation_manager"
data-ga-label="save">
Create file
</button>
<button class="action-panel-cancel"
aria-label="Close 'Send citations to citation manager' panel"
ref="linksrc=close_citation_manager_panel"
aria-controls="citation-manager-action-panel"
aria-expanded="false"
data-ga-category="save_share"
data-ga-action="citation_manager"
data-ga-label="cancel">
Cancel
</button>
</div>
</form>
</div>
</div>
<div id="rss-action-panel" class="rss-action-panel action-panel " aria-hidden="true">
<div class="inner-wrap">
<h2 class="action-panel-heading">
Your RSS Feed
</h2>
<form id="rss-action-panel-form"
class="rss-action-panel-form action-panel-content action-form"
data-create-rss-feed-url="/create-rss-feed-url/"
data-search-form-term-value="">
<input type="hidden" name="csrfmiddlewaretoken" value="tjvTMg4THnEvebrvKI4ncRNOvsmbwWd9K6fYp6xfhYrZ8Cxln1HpGZjs3w4npxLt">
<div class="action-panel-control-wrap">
<label for="rss-name" class="action-panel-label required-field-asterisk">
Name of RSS Feed:
</label>
<input maxlength="200"
placeholder="Name"
type="text"
name="rss-name"
id="rss-name"
class="rss-name"
value=''
required
pattern="[^&quot;&amp;=&lt;&gt;\/]*" title="The following characters are not allowed in the Name field: &quot;&amp;=&lt;&gt;/">
</div>
<div class="rss-limit-wrap">
<div class="action-panel-control-wrap action-panel-extra-margin-top">
<label for="rss-limit" class="action-panel-label">
Number of items displayed:
</label>
<select id="rss-limit" class="no-border-panel-selector rss-limit">
<option value="5">5</option>
<option value="10">10</option>
<option value="15" selected="selected">15</option>
<option value="20">20</option>
<option value="50">50</option>
<option value="100">100</option>
</select>
</div>
</div>
<div class="action-panel-actions">
<button class="action-panel-submit"
type="submit"
data-loading-label="Creating..."
data-ga-category="save_share"
data-ga-action="alert"
data-ga-label="save">
Create RSS
</button>
<button class="action-panel-cancel"
aria-label="Close 'Your RSS' panel"
ref="linksrc=close_rss_panel"
aria-controls="rss-action-panel"
aria-expanded="false"
data-ga-category="save_share"
data-ga-action="alert"
data-ga-label="cancel">
Cancel
</button>
</div>
<div class="action-panel-control-wrap rss-link-copy-wrap">
<label for="rss-link" class="usa-sr-only">RSS Link</label>
<input placeholder="Your RSS Feed Link" type="text" name="rss-link" id="rss-link" class="rss-link" title="RSS Link">
<button
type="button"
disabled
class="rss-link-copy-button disabled"
data-ga-category="save_share"
data-ga-action="rss"
data-ga-label="copy">
Copy
</button>
</div>
</form>
</div>
</div>
</div>
</div>
<div class="article-page" id="article-page" data-article-pmid="33754048">
<aside class="page-sidebar">
<div class="inner-wrap">
<div class="full-text-links">
<div class="full-view">
<h3 class="title">
Full text links
</h3>
<div class="full-text-links-list">
<a class="link-item
dialog-focus"
href="https://www.thno.org/v11p4061.htm"
target="_blank"
rel="noopener"
ref="linksrc=fulltextorjournal_fulltext&amp;is_pmc=False&amp;PrId=6264&amp;itool=Abstract-def&amp;log$=linkouticon&amp;uid=33754048&amp;db=pubmed&amp;nlmid=101552395"
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Ivyspring International Publisher
</span></a><a class="link-item
pmc
"
href="https://pmc.ncbi.nlm.nih.gov/articles/pmid/33754048/"
target="_blank"
rel="noopener"
ref="linksrc=fulltextorjournal_fulltext&amp;is_pmc=True&amp;PrId=3494&amp;itool=Abstract-def&amp;log$=linkouticon&amp;uid=33754048&amp;db=pubmed&amp;nlmid=101552395"
title="Free full text at PubMed Central"
data-ga-category="full_text"
data-ga-action="PMC"
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Free PMC article
</span></a>
</div>
</div>
<div class="short-view">
<a href="#" class="full-text-links-button full-text-links-dialog-trigger">
Full text links
</a>
</div>
</div>
<div class="actions-buttons sidebar"><h3 class="title">Actions</h3><div class="inner-wrap"><button class="citation-button citation-dialog-trigger"
aria-label="Open dialog with citation text in different styles"
data-ga-category="save_share"
data-ga-action="cite"
data-ga-label="open"
data-all-citations-url="/33754048/citations/"
data-citation-style="nlm"
data-pubmed-format-link="/33754048/export/"><span class="button-label">Cite</span></button><link type="text/css" href="ncbi-overlay-block/src/overlay-block.css"><div class="collections-button-container" data-article-id="33754048" data-article-db="pubmed"><button class="collections-button collections-dialog-trigger"
aria-label="Save article in MyNCBI collections."
data-ga-category="collections_button"
data-ga-action="click"
data-ga-label="collections_button"
data-collections-open-dialog-enabled="false"
data-collections-open-dialog-url="https://account.ncbi.nlm.nih.gov/?back_url=https%3A%2F%2Fpubmed.ncbi.nlm.nih.gov%2F33754048%2F%23open-collections-dialog"
data-in-collections="false"><span class="button-label">Collections</span></button><div class="overlay" role="dialog"><div id="collections-action-dialog"
class="dialog collections-dialog"
aria-hidden="true"><div class="title">Add to Collections</div><div class="collections-action-panel action-panel"><form id="collections-action-dialog-form"
class="collections-action-panel-form action-panel-content action-form action-panel-smaller-selectors"
data-existing-collections-url="/list-existing-collections/"
data-add-to-existing-collection-url="/add-to-existing-collection/"
data-create-and-add-to-new-collection-url="/create-and-add-to-new-collection/"
data-myncbi-max-collection-name-length="100"
data-collections-root-url="https://www.ncbi.nlm.nih.gov/myncbi/collections/"><input type="hidden" name="csrfmiddlewaretoken" value="tjvTMg4THnEvebrvKI4ncRNOvsmbwWd9K6fYp6xfhYrZ8Cxln1HpGZjs3w4npxLt"><div class="choice-group" role="radiogroup"><ul class="radio-group-items"><li><input type="radio"
id="collections-action-dialog-new"
class="collections-new"
name="collections"
value="new"
data-ga-category="collections_button"
data-ga-action="click"
data-ga-label="collections_radio_new"><label for="collections-action-dialog-new">Create a new collection</label></li><li><input type="radio"
id="collections-action-dialog-existing"
class="collections-existing"
name="collections"
value="existing"
checked="true"
data-ga-category="collections_button"
data-ga-action="click"
data-ga-label="collections_radio_existing"><label for="collections-action-dialog-existing">Add to an existing collection</label></li></ul></div><div class="controls-wrapper"><div class="action-panel-control-wrap new-collections-controls"><label for="collections-action-dialog-add-to-new" class="action-panel-label required-field-asterisk">
Name your collection:
</label><input
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miR-375 prevents high-fat diet-induced insulin resistance and obesity by targeting the aryl hydrocarbon receptor and bacterial tryptophanase (<i>tnaA</i>) gene
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miR-375 prevents high-fat diet-induced insulin resistance and obesity by targeting the aryl hydrocarbon receptor and bacterial tryptophanase (<i>tnaA</i>) gene
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<b>Background:</b> Diet manipulation is the basis for prevention of obesity and diabetes. The molecular mechanisms that mediate the diet-based prevention of insulin resistance are not well understood. Here, as proof-of-concept, ginger-derived nanoparticles (GDNP) were used for studying molecular mechanisms underlying GDNP mediated prevention of high-fat diet induced insulin resistance. <b>Methods:</b> Ginger-derived nanoparticles (GDNP) were isolated from ginger roots and administered orally to C57BL/6 high-fat diet mice. Fecal exosomes released from intestinal epithelial cells (IECs) of PBS or GDNP treated high-fat diet (HFD) fed mice were isolated by differential centrifugation. A micro-RNA (miRNA) polymerase chain reaction (PCR) array was used to profile the exosomal miRs and miRs of interest were further analyzed by quantitative real time (RT) PCR. miR-375 or antisense-miR375 was packed into nanoparticles made from the lipids extracted from GDNP. Nanoparticles was fluorescent labeled for monitoring their <i>in vivo</i> trafficking route after oral administration. The effect of these nanoparticles on glucose and insulin response of mice was determined by glucose and insulin tolerance tests. <b>Results:</b> We report that HFD feeding increased the expression of AhR and inhibited the expression of miR-375 and VAMP7. Treatment with orally administered ginger-derived nanoparticles (GDNP) resulted in reversing HFD mediated inhibition of the expression of miR-375 and VAMP7. miR-375 knockout mice exhibited impaired glucose homeostasis and insulin resistance. Induction of intracellular miR-375 led to inhibition of the expression of AhR and VAMP7 mediated exporting of miR-375 into intestinal epithelial exosomes where they were taken up by gut bacteria and inhibited the production of the AhR ligand indole. Intestinal exosomes can also traffic to the liver and be taken up by hepatocytes, leading to miR-375 mediated inhibition of hepatic AhR over-expression and inducing the expression of genes associated with the hepatic insulin response. Altogether, GDNP prevents high-fat diet-induced insulin resistance by miR-375 mediated inhibition of the aryl hydrocarbon receptor mediated pathways over activated by HFD feeding. <b>Conclusion:</b> Collectively our findings reveal that oral administration of GDNP to HFD mice improves host glucose tolerance and insulin response via regulating AhR expression by GDNP induced miR-375 and VAMP7.
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AhR; E. coli tryptophanase (tnaA); Exosomes; Ginger derived nanoparticles; VAMP7; gut/liver axis; indole; insulin resistance.; miR-375.
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<div class="figure-caption-contents"><b> miR-375 regulates AhR expression. (A)… </b></div>
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<div class="figure-caption-contents"><b> miR-375 regulates AhR expression. (A) </b> Bar graph showing the fold-change in the expression…</div>
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Figure 1
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<div class="figure-caption-contents"><b>miR-375 regulates AhR expression. (A)</b> Bar graph showing the fold-change in the expression of microRNAs in HFD mice small intestinal tissue induced by GDNP treatment, (expression following treatment with PBS served as the baseline value). The bar graph shows only microRNAs with &gt;5 fold up- or downregulation. <b>(B)</b> Alteration of gene expression of small intestine (SI) tissues from high-fat diet (HFD)-fed mice treated with either PBS or GDNP (using an Affymetrix array). Red boxes highlighted are the genes involved in insulin signaling and lipid metabolism. <b>(C)</b> qPCR assay for miR-375 in cells cultured with PBS or GDNP for 12 h. <b>(D)</b> Graphical representation of the miR-375 binding site in AhR mRNA as predicted by TargetScan. <b>(E)</b> qPCR quantification of <i>AhR</i> expression in small intestine tissues of HFD-fed mice treated with PBS or GDNP. <b>(F)</b> Western blot presenting AhR expression in the small intestine tissues of lean and HFD-fed mice treated with PBS or GDNP. <b>(G &amp; H)</b>
<i>AhR</i> mRNA by qPCR (G) and protein expression by western blot (H) in MC-38 cells treated with PBS or GDNP with anti-sense-miR375 or scramble. Ratio to β-actin were shown (numbers in the middle of each panel).<b> (I &amp; J)</b>
<i>AhR</i> mRNA (I) and protein expression (J) in the small intestine tissues of RCD mice orally administered nanoparticles (Nano-scramble or Nano-miR375). Ratio to β-actin is shown in the middle as numbers. One-way ANOVA with the Bonferroni correction for multiple comparisons and the Student <i>t</i> test (one tailed) were used to calculate statistical significance (<i>p</i> value *&lt;0.05; **&lt;0.01; ***&lt;0.001; ****&lt;0.0001).</div>
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<div class="figure-caption-contents"><b> GDNP induces miR-375 in MC-38… </b></div>
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<p> Figure 2 </p>
</strong>
<div class="figure-caption-contents"><b> GDNP induces miR-375 in MC-38 cells and sorts into exosomes. (A) </b> qRT-PCR for…</div>
</div>
<figcaption id="figure-caption-1" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
Figure 2
</strong>
<div class="figure-caption-contents"><b> GDNP induces miR-375 in MC-38 cells and sorts into exosomes. (A)</b> qRT-PCR for intracellular and exosomal miR-375 levels and AhR expression in MC-38 cells cultured with various concentrations of GDNP for 12 h. <b>(B)</b> qRT-PCR comparison for intracellular vs exosomal levels of miR-375 in MC-38 cells cultured with various concentrations of GDNP. <b>(C)</b> miR-375 levels expression in MC-38 cells cultured with GDNP in a time dependent manner. <b>(D)</b> miR-375 levels expression in FL83B (hepatocyte) cells cultured with GDNP in a dose dependent manner. One-way ANOVA with the Bonferroni correction for multiple comparisons and the Student <i>t</i> test (one tailed) were used to calculate statistical significance (<i>p</i> value *&lt;0.05; **&lt;0.01; ***&lt;0.001).</div>
</figcaption>
</figure>
<figure class="figure-item "
itemscope itemtype="http://schema.org/ImageObject"
itemprop="associatedMedia"
data-slide-index="2"
data-label-slug="figure-3">
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href="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/00fa/7977461/6342d70d6bec/thnov11p4061g003.jpg"
itemprop="contentUrl"
aria-describedby="figure-caption-2"
role="button"
data-image-width="794"
data-image-height="1038"
data-image-alt="Figure 3"
data-pmc-id="PMC7977461"
data-figure-id="F3">
<img class="figure-thumb" itemprop="thumbnail"
id="article-image-2"
src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/00fa/7977461/15fa29cd6123/thnov11p4061g003.gif"
alt="Figure 3" />
</a>
<meta itemprop="width" itemtype="http://schema.org/ImageObject" content="794">
<meta itemprop="height" itemtype="http://schema.org/ImageObject" content="1038">
<div class="figure-caption figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="caption" aria-hidden="true">
<div class="caption-wrap">
<strong class="figure-label">
<p> Figure 3 </p>
</strong>
<div class="figure-caption-contents"><b> miR-375 is sorted into exosomes… </b></div>
</div>
</div>
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> Figure 3 </p>
</strong>
<div class="figure-caption-contents"><b> miR-375 is sorted into exosomes via VAMP7 and intracellular miR-375 regulates AhR expression.… </b></div>
</div>
<figcaption id="figure-caption-2" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
Figure 3
</strong>
<div class="figure-caption-contents"><b> miR-375 is sorted into exosomes via VAMP7 and intracellular miR-375 regulates AhR expression. (A)</b> Western blots for VAMP7 protein levels in MC-38 cells cultured with various concentrations of GDNP. Ratio of VAMP7 to β-actin is shown as a number between the two gels. <b>(B)</b> miRNA array expression profile of intestinal epithelial cell exosomes (A33<sup>+</sup>CD63<sup>+</sup> exosomes) from feces derived from HFD-fed mice treated with GDNP vs PBS. <b>(C)</b> Bar graph showing HFD-fed mouse fecal exosomal miRNAs with a fold change (&gt;25-fold or &lt;5-fold) following treatment with GDNP vs PBS. <b>(D)</b> qRT PCR analysis of miR-375 expression in intestinal epithelial cell exosomes (A33<sup>+</sup>CD63<sup>+</sup>) from lean and HFD-fed mice treated with GDNP vs PBS. <b>(E)</b> Western blot showing VAMP7 expression in the small intestine of lean and HFD-fed mice treated with PBS or GDNP. Ratio of VAMP7 to β-actin is shown as a number between the two panels. <b>(F &amp; G)</b> qRT-PCR analysis (F) and western blot (G) of VAMP7 expression in PBS- or GDNP-treated MC-38 cells. <b>(H)</b> Confocal images displaying VAMP7 expression in PBS- or GDNP-treated MC-38 cells. <b>(I)</b> qPCR analysis of the intracellular expression of miR-375 in WT and VAMP7KO MC-38 cells. <b>(J)</b> qPCR analysis of the exosomal levels of miR-375 in WT and VAMP7KO MC-38 cells. <b>(K)</b> MC-38 cells were transfected with biotinylated miR-375 and pulled-down with streptavidin beads. Western blots were carried out for VAMP7 using eluted extract from streptavidin beads. <b>(L)</b> qPCR for miR-375 in pulled-down product used for cDNA preparation. One-way ANOVA with the Bonferroni correction for multiple comparisons and the Student <i>t</i> (two-tailed) test were used to calculate statistical significance (<i>p</i> value *&lt;0.05; **&lt;0.01; ***&lt;0.001).</div>
</figcaption>
</figure>
<figure class="figure-item "
itemscope itemtype="http://schema.org/ImageObject"
itemprop="associatedMedia"
data-slide-index="3"
data-label-slug="figure-4">
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itemprop="contentUrl"
aria-describedby="figure-caption-3"
role="button"
data-image-width="755"
data-image-height="485"
data-image-alt="Figure 4"
data-pmc-id="PMC7977461"
data-figure-id="F4">
<img class="figure-thumb" itemprop="thumbnail"
id="article-image-3"
src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/00fa/7977461/b0841c90ea4d/thnov11p4061g004.gif"
alt="Figure 4" />
</a>
<meta itemprop="width" itemtype="http://schema.org/ImageObject" content="755">
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<div class="figure-caption figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="caption" aria-hidden="true">
<div class="caption-wrap">
<strong class="figure-label">
<p> Figure 4 </p>
</strong>
<div class="figure-caption-contents"><b> Gut epithelial cell exosomes (CD63… </b></div>
</div>
</div>
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> Figure 4 </p>
</strong>
<div class="figure-caption-contents"><b> Gut epithelial cell exosomes (CD63 <sup> + </sup> A33 <sup> + </sup> ) influence gut bacterial populations… </b></div>
</div>
<figcaption id="figure-caption-3" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
Figure 4
</strong>
<div class="figure-caption-contents"><b> Gut epithelial cell exosomes (CD63<sup>+</sup>A33<sup>+</sup>) influence gut bacterial populations and modulate microbial metabolites. (A)</b> Representative electron micrograph of gut bacteria containing fecal exosomes<b>.</b> Yellow arrows indicate exosomes inside and outside the bacteria. <b>(B)</b> FACS analysis of PKH26-positive gut bacteria from mice orally administered PKH-26-labeled fecal exosomes. <b>(C)</b> Confocal images of bacteria showing uptake of PKH-26-labeled fecal exosomes (red). <b>(D)</b> Schematic diagram of the putative targeting site of mmu-miR-375-3p in the <i>E. coli tryptophanase</i> (<i>tnaA</i>). <b>(E)</b> mRNA levels of the <i>tnaA</i> gene in gut bacteria derived from HFD mice treated with PBS or GDNP. <b>(F)</b> 2D LC-MS/MS analysis of unmetabolized tryptophan levels excreted into the feces of PBS- or GDNP-treated HFD-fed mice. <b>(G)</b> Quantification of the indole levels in the feces (h) and plasma (i) obtained from lean and HFD-fed mice that were treated with PBS or GDNP. <b>(H)</b> Fold change in<i> tnaA</i> gene expression in fecal bacteria (left), and indole estimation in the fecal supernatants (middle) and plasma (right) from GDNP-treated HFD-fed mice treated with PBS or nanoparticles packaged with antisense-miR375 or scramble. One-way ANOVA with the Tukey correction for multiple comparisons and the Student <i>t</i> (one tailed) test were used to calculate statistical significance (<i>p</i> value *&lt;0.05; **&lt;0.01; ***&lt;0.001; ****&lt;0.0001).</div>
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data-label-slug="figure-5">
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itemprop="contentUrl"
aria-describedby="figure-caption-4"
role="button"
data-image-width="758"
data-image-height="397"
data-image-alt="Figure 5"
data-pmc-id="PMC7977461"
data-figure-id="F5">
<img class="figure-thumb" itemprop="thumbnail"
id="article-image-4"
src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/00fa/7977461/d39263b7b97d/thnov11p4061g005.gif"
alt="Figure 5" />
</a>
<meta itemprop="width" itemtype="http://schema.org/ImageObject" content="758">
<meta itemprop="height" itemtype="http://schema.org/ImageObject" content="397">
<div class="figure-caption figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="caption" aria-hidden="true">
<div class="caption-wrap">
<strong class="figure-label">
<p> Figure 5 </p>
</strong>
<div class="figure-caption-contents"><b> Human fecal exosomal miR-375 is… </b></div>
</div>
</div>
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> Figure 5 </p>
</strong>
<div class="figure-caption-contents"><b> Human fecal exosomal miR-375 is negatively correlated with indole production. (A) </b> qPCR analysis…</div>
</div>
<figcaption id="figure-caption-4" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
Figure 5
</strong>
<div class="figure-caption-contents"><b> Human fecal exosomal miR-375 is negatively correlated with indole production. (A)</b> qPCR analysis of miR-375 levels in human fecal exosomes and plasma exosomes derived from healthy, obese and individuals with T2D. <b>(B)</b> Quantification of indole levels in the feces and plasma. <b>(C)</b> Quantification of plasma cholesterol and triglyceride levels. <b>(D)</b> Scatter plot depicting the linear correlation between cholesterol and miR-375 levels, and triglycerides and miR-375 levels. <b>(E)</b> Principle component analysis (PCoA) of miR-375 and indole levels in obese, T2D and healthy human fecal exosomes. One-way ANOVA with the Bonferroni correction for multiple comparisons was used to calculate statistical significance. (<i>p</i> value *&lt;0.05; **&lt;0.01; ***&lt;0.001).</div>
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data-slide-index="5"
data-label-slug="figure-6">
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itemprop="contentUrl"
aria-describedby="figure-caption-5"
role="button"
data-image-width="762"
data-image-height="920"
data-image-alt="Figure 6"
data-pmc-id="PMC7977461"
data-figure-id="F6">
<img class="figure-thumb" itemprop="thumbnail"
id="article-image-5"
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<meta itemprop="width" itemtype="http://schema.org/ImageObject" content="762">
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<strong class="figure-label">
<p> Figure 6 </p>
</strong>
<div class="figure-caption-contents"><b> miR-375 improves insulin sensitivity and… </b></div>
</div>
</div>
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> Figure 6 </p>
</strong>
<div class="figure-caption-contents"><b> miR-375 improves insulin sensitivity and glucose homeostasis and prevents dyslipidemia. (A) </b> Graphical representation…</div>
</div>
<figcaption id="figure-caption-5" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
Figure 6
</strong>
<div class="figure-caption-contents"><b> miR-375 improves insulin sensitivity and glucose homeostasis and prevents dyslipidemia. (A)</b> Graphical representation of the experiment, which consisted of adoptive transfer of CD63<sup>+</sup>A33<sup>+</sup> fecal exosomes (HFD-Exo) from HFD mouse (HFD fed 12 months) plus nanoparticles containing miR-375. Nanoparticles generated using the total lipid from GDNP. <b>(B)</b> Live imaging of mice orally administered PKH26 labeled nanoparticles containing miR-375. <b>(C)</b> Imaging of the liver, small and large intestines indicating the presence of labeled nanoparticles 6 hours after oral administration. <b>(D)</b> PKH26 labeled nanoparticle uptake by hepatocytes (albumin-positive cells) or Kupffer (F4/80-positive) cells. <b>(E)</b> Representative images of cellular uptake of PKH26-labeled nanoparticles by hepatocytes (albumin-positive cells). PKH26-labeled particles are indicated by pink arrows. <b>(F)</b> 3D image of PKH26-labeled nanoparticles in hepatocytes. <b>(G)</b> Confocal imaging to detect AhR (FITC) and biotinylated miR-375 or scrambled microRNA in liver tissues derived from mice orally administered nanoparticles. <b>(H)</b> GTT and ITT for C57BL/6 mice that received the fecal exosomes (HFD-Exo) along with nanoparticles containing miR-375 or scrambled RNA for 14 days while the mice were fed a HFD. Statistical comparisons were made between HFD-Exo vs Nano-miR375; HFD-Exo was responsible for insulin resistance and miR-375 responsible for preventing the development of insulin resistance. Nanoparticles contained scramble RNA (Nano-scramble); nanoparticles only (Nano); and nanoparticles contained miR-375 (Nano-miR375). <b>(I)</b> Cholesterol and triglyceride levels in plasma derived from HFD mice treated with either PBS or nanoparticles (above mentioned) for 14 days. <b>(J)</b> Insulin signaling array of mouse hepatocytes cultured with fecal exosomes (HFD-Exo) along with nanoparticles (contained scramble &amp; nano-miR-375) showing alterations in genes involved in insulin signaling. Green-boxed genes promote insulin activity and red-boxed genes inhibit insulin activity. <b>(K)</b> Western blot depicting Foxa2, AhR, and IRS-1 and 2 expression in hepatocytes treated with fecal exosomes (HFD-Exo) along with nanoparticles (contained scramble &amp; nano-miR-375). The ratio to β-actin is shown in bar graph form as part of panel K. <b>(L)</b> Effect of fecal exosomes (HFD-Exo) on glucose uptake by hepatocytes. One-way ANOVA with a Bonferroni correction for multiple comparisons test was used to calculate statistical significance. (<i>p</i> value *&lt;0.05; **&lt;0.01; ***&lt;0.001; ****&lt;0.0001).</div>
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<span class="docsum-journal-citation full-journal-citation">Front Nutr. 2023 Jul 13;10:1223349. doi: 10.3389/fnut.2023.1223349. eCollection 2023.</span>
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