Name: use of animal model of sepsis to evaluate novel herbal therapies
Uploaded: 2012-04-11T13:00:00
Description: Watch this Scientific Journal Video about Use of Animal Model of Sepsis to Evaluate Novel Herbal Therapies at JoVE.com

This work was supported by grants from the National Institutes of Health, National Institute of General Medical Science (R01GM063075) and the National Center of Complementary and Alternative Medicine (R01AT05076).

1. Establishment of Animal Model of Sepsis
<ol>
	<li>Mice are anesthetized with ketamine (75 mg/kg, intramuscular, i.m.) and xylazine (10 mg/kg, i.m.), and placed in supine position.</li>
	<li>Fix the feet of the mouse with tape to ensure a stable position.</li>
	<li>Clean the abdomen with 3 alternating scrubs of betadine or other skin disinfectant and alcohol. Then, make a 15 mm midline incision to expose the cecum.</li>
	<li>Ligate the cecum with a 4-0 silk suture at approximately 5.0 mm from the cecal tip, and then...

<ol>
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In the laboratory, several animal models of sepsis have been employed to understand the pathogenesis of sepsis in order to develop potential novel therapies. Their clinical relevance remains a subject of debate before the successful translation of animal studies into clinical applications for sepsis. Although neutralizing antibodies against early cytokines (e.g., TNF) were protective in animal models of bacteremia/endotoxemia17,18, they actually worsen survival in animal model of sepsis19. Similarly...

<table border="1">
 <tbody>
 <tr>
 <td>Name of the reagent</td>
 <td>Company</td>
 <td>Catalogue number</td>
 </tr>
 <tr>
 <td>Betadine </td>
 <td>Purdue Products L.P.</td>
 <td>25655-41-8</td>
 </tr>
 <tr>
 <td>imipenem</td>
 <td>Merck &amp; Co., Inc.</td>
 <td>9882821</td>
 </tr>
 <tr>
 <td>Ketamine HCl</td>
 <td>Hospira Inc.</td>
 <td>RL-0065</td>
 </tr>
 <tr>
 <td>Xylazine</td>
 <td>Lloyd Laboratories</td>
 <td>4821</td>
 </tr>
 <tr>
 <td>Autoclip </td>
 <td>Becton Dickinson</td>
 <td>427631</td>
 </tr>
 <tr>
 <td>4-0 silk suture</td>
 <td>Roboz</td>
 <td>SUT-15-2</td>
 </tr>
 <tr>
 <td>Surflo I.V. Catheter</td>
 <td>Terumo</td>
 <td>SR*OX2419CA</td>
 </tr>
 <tr>
 <td>RayBio mouse cytokine antibody array</td>
 <td>RayBiotech, Inc.</td>
 <td>AAM-CYT-3</td>
 </tr>
 <tr>
 <td>Thioglycollate </td>
 <td>Becton Dickinson</td>
 <td>211716</td>
 </tr>
 </tbody>
</table>

use of animal model of sepsis to evaluate novel herbal therapies

Sepsis refers to a systemic inflammatory response syndrome resulting from a microbial infection. It has been routinely simulated in animals by several techniques, including infusion of exogenous bacterial toxin (endotoxemia) or bacteria (bacteremia), as well as surgical perforation of the cecum by cecal ligation and puncture (CLP)1-3. CLP allows bacteria spillage and fecal contamination of the peritoneal cavity, mimicking the human clinical disease of perforated appendicitis or diverticulitis. The severity of sepsis, as reflected by the eventual mortality rates, can be controlled surgically by varying the size of the needle used for cecal puncture2. In animals, CLP induces similar, biphasic hemodynamic cardiovascular, metabolic, and immunological responses as observed during the clinical course of human sepsis3. Thus, the CLP model is considered as one of the most clinically relevant models for experimental sepsis1-3.
Various animal models have been used to elucidate the intricate mechanisms underlying the pathogenesis of experimental sepsis. The lethal consequence of sepsis is attributable partly to an excessive accumulation of early cytokines (such as TNF, IL-1 and IFN-&gamma;)4-6 and late proinflammatory mediators (e.g., HMGB1)7. Compared with early proinflammatory cytokines, late-acting mediators have a wider therapeutic window for clinical applications. For instance, delayed administration of HMGB1-neutralizing antibodies beginning 24 hours after CLP, still rescued mice from lethality8,9, establishing HMGB1 as a late mediator of lethal sepsis. The discovery of HMGB1 as a late-acting mediator has initiated a new field of investigation for the development of sepsis therapies using Traditional Chinese Herbal Medicine. In this paper, we describe a procedure of CLP-induced sepsis, and its usage in screening herbal medicine for HMGB1-targeting therapies.

Watch this Scientific Journal Video about Use of Animal Model of Sepsis to Evaluate Novel Herbal Therapies at JoVE.com

Use of Animal Model of Sepsis to Evaluate Novel Herbal Therapies

Sepsis refers to a systemic inflammatory response syndrome resulting from a microbial infection, and can be simulated by a surgical technique termed cecal ligation and puncture (CLP). Here we describe a method to use CLP-induced animal model to screen medicinal herbs for therapeutic agents.

Sepsis refers to a systemic inflammatory response syndrome resulting from a microbial infection. It has been routinely simulated in animals by several ...

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