<meta charset="utf8"></head><body>Hindlimb suspension을 이용한 패혈증 유발 근병증을 연구하기 위한 마우스 모델

<ol>
	<li>Prescott, H. C., Angus, D. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Enhancing+recovery+from+sepsis.">Enhancing recovery from sepsis.</a> JAMA. 319 (1), 62-75 (2018).</li><li>Stortz, J. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Benchmarking+clinical+outcomes+and+the+immunocatabolic+phenotype+of+chronic+critical+illness+after+sepsis+in+surgical+intensive+care+unit+patients.">Benchmarking clinical outcomes and the immunocatabolic phenotype of chronic critical illness after sepsis in surgical intensive care unit patients.</a> J Trauma Acute Care Surg. 84 (2), 342-349 (2018).</li><li>Laitano, O., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+impact+of+hindlimb+disuse+on+sepsis-induced+myopathy+in+mice.">The impact of hindlimb disuse on sepsis-induced myopathy in mice.</a> Physiological Rep. 9 (14), e14979 (2021).</li><li>Callahan, L. A., Supinski, G. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sepsis-induced+myopathy.">Sepsis-induced myopathy.</a> Crit Care Med. 37, S354-S367 (2009).</li><li>Cuthbertson, B. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mortality+and+quality+of+life+in+the+five+years+after+severe+sepsis.">Mortality and quality of life in the five years after severe sepsis.</a> Crit Care. 17 (2), R70 (2013).</li><li>Schmitt, R. E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Muscle+stem+cells+contribute+to+long-term+tissue+repletion+following+surgical+sepsis.">Muscle stem cells contribute to long-term tissue repletion following surgical sepsis.</a> J Cachexia Sarcopenia Muscle. 14 (3), 1424-1440 (2023).</li><li>Owen, A. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chronic+muscle+weakness+and+mitochondrial+dysfunction+in+the+absence+of+sustained+atrophy+in+a+preclinical+sepsis+model.">Chronic muscle weakness and mitochondrial dysfunction in the absence of sustained atrophy in a preclinical sepsis model.</a> eLife. 8, e49920 (2019).</li><li>Chen, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cellular+senescence+implicated+in+sepsis-induced+muscle+weakness+and+ameliorated+with+metformin.">Cellular senescence implicated in sepsis-induced muscle weakness and ameliorated with metformin.</a> Shock. 59 (4), 646 (2023).</li><li>Granger, J. I., Ratti, P. L., Datta, S. C., Raymond, R. M., Opp, M. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sepsis-induced+morbidity+in+mice:+effects+on+body+temperature,+body+weight,+cage+activity,+social+behavior+and+cytokines+in+brain.">Sepsis-induced morbidity in mice: effects on body temperature, body weight, cage activity, social behavior and cytokines in brain.</a> Psychoneuroendocrinology. 38 (7), 1047-1057 (2013).</li><li>Ruiz, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sepsis+modeling+in+mice:+ligation+length+is+a+major+severity+factor+in+cecal+ligation+and+puncture.">Sepsis modeling in mice: ligation length is a major severity factor in cecal ligation and puncture.</a> Intensive Care Med Exp. 4 (1), 22 (2016).</li><li>Laitano, O., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Xiphoid+surface+temperature+predicts+mortality+in+a+murine+model+of+septic+shock.">Xiphoid surface temperature predicts mortality in a murine model of septic shock.</a> Shock (Augusta, Ga). 50 (2), 226-232 (2018).</li><li>Park, S., Brisson, B. K., Liu, M., Spinazzola, J. M., Barton, E. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mature+IGF-I+excels+in+promoting+functional+muscle+recovery+from+disuse+atrophy+compared+with+pro-IGF-IA.">Mature IGF-I excels in promoting functional muscle recovery from disuse atrophy compared with pro-IGF-IA.</a> J App Physiol. 116 (7), 797-806 (2013).</li><li>Spradlin, R. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Deletion+of+muscle+Igf1+exacerbates+disuse+atrophy+weakness+in+mice.">Deletion of muscle Igf1 exacerbates disuse atrophy weakness in mice.</a> J App Physiol. 131 (3), 881-894 (2021).</li><li>Shrum, B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+robust+scoring+system+to+evaluate+sepsis+severity+in+an+animal+model.">A robust scoring system to evaluate sepsis severity in an animal model.</a> BMC Res Notes. 7 (1), 233 (2014).</li><li>Mai, S. H. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Body+temperature+and+mouse+scoring+systems+as+surrogate+markers+of+death+in+cecal+ligation+and+puncture+sepsis.">Body temperature and mouse scoring systems as surrogate markers of death in cecal ligation and puncture sepsis.</a> Intensive Care Med Exp. 6 (1), 20 (2018).</li><li>Vankrunkelsven, W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+muscle+weakness+in+a+mouse+model+of+critical+illness:+does+fibroblast+growth+factor+21+play+a+role.">Development of muscle weakness in a mouse model of critical illness: does fibroblast growth factor 21 play a role.</a> Skelet Muscle. 13 (1), 12 (2023).</li><li>Supinski, G., Stofan, D., Nethery, D., Szweda, L., DiMarco, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Apocynin+improves+diaphragmatic+function+after+endotoxin+administration.">Apocynin improves diaphragmatic function after endotoxin administration.</a> J Appl Physiol. 87 (2), 776-782 (1999).</li><li>Li, X., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sepsis+leads+to+impaired+mitochondrial+calcium+uptake+and+skeletal+muscle+weakness+by+reducing+the+micu1:mcu+protein+ratio.">Sepsis leads to impaired mitochondrial calcium uptake and skeletal muscle weakness by reducing the micu1:mcu protein ratio.</a> Shock (Augusta, Ga). 60 (5), 698-706 (2023).</li><li>Wang, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Targeting+NAT10+protects+against+sepsis-induced+skeletal+muscle+atrophy+by+inhibiting+ROS/NLRP3.">Targeting NAT10 protects against sepsis-induced skeletal muscle atrophy by inhibiting ROS/NLRP3.</a> Life Sci. 330, 121948 (2023).</li><li>Mankowski, R. T., Laitano, O., Clanton, T. L., Brakenridge, S. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pathophysiology+and+treatment+strategies+of+acute+myopathy+and+muscle+wasting+after+sepsis.">Pathophysiology and treatment strategies of acute myopathy and muscle wasting after sepsis.</a> J Clinical Med. 10 (9), 1874 (2021).</li><li>Oliveira, J. R. S., Mohamed, J. S., Myers, M., Brooks, M. J., Alway, S. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+hindlimb+suspension+and+reloading+on+gastrocnemius+and+soleus+muscle+mass+and+function+in+geriatric+mice.">Effects of hindlimb suspension and reloading on gastrocnemius and soleus muscle mass and function in geriatric mice.</a> Exp Gerontol. 115, 19-31 (2019).</li><li>Ashare, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Insulin-like+growth+factor-1+improves+survival+in+sepsis+via+enhanced+hepatic+bacterial+clearance.">Insulin-like growth factor-1 improves survival in sepsis via enhanced hepatic bacterial clearance.</a> Am J Respir Crit Care Med. 178 (2), 149-157 (2008).</li><li>Starr, M. E., Saito, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Age-related+increase+in+food+spilling+by+laboratory+mice+may+lead+to+significant+overestimation+of+actual+food+consumption:+implications+for+studies+on+dietary+restriction,+metabolism,+and+dose+calculations.">Age-related increase in food spilling by laboratory mice may lead to significant overestimation of actual food consumption: implications for studies on dietary restriction, metabolism, and dose calculations.</a> J Gerontol A Biol Sci Med Sci. 67 (10), 1043-1048 (2012).</li><li>Crowell, K. T., Soybel, D. I., Lang, C. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Restorative+mechanisms+regulating+protein+balance+in+skeletal+muscle+during+recovery+from+sepsis.">Restorative mechanisms regulating protein balance in skeletal muscle during recovery from sepsis.</a> Shock. 47 (4), 463-473 (2017).</li></ol>

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>4-0 Ethicon Coated Vicryl</td>
			<td>Ethicon</td>
			<td>D5792</td>
			<td>Absorbable suture used for closure of muscle layer and for ligation of the cecum.</td>
		</tr>
		<tr>
			<td>4-0 Ethilon Black 18&#34;&#160;</td>
			<td>Ethicon</td>
			<td>662G</td>
			<td>Non absorbable suture for closure of the skin layer.</td>
		</tr>
		<tr>
			<td>BD&#160; PrecisionGlide Needle 26-28 G</td>
			<td>BD</td>
			<td>305136 for 27g needle</td>
			<td>Needle for puncturing the cecum.</td>
		</tr>
		<tr>
			<td>C57BL/6J mice&#160;</td>
			<td>Jackson Laboratory&#160;</td>
			<td>strain #000664</td>
			<td></td>
		</tr>
		<tr>
			<td>Cotton Tipped Applicators</td>
			<td>Puritan</td>
			<td>S-18991</td>
			<td>Swabs for topical application of iodine.</td>
		</tr>
		<tr>
			<td>Cryostat</td>
			<td>(Leica CM1950)</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Dynarex Povidone Iodine Prep Solution</td>
			<td>Dynarex</td>
			<td>1415</td>
			<td>Topical Antiseptic Liquid for Skin and Mucosa</td>
		</tr>
		<tr>
			<td>Ethanol 200 Proof (100%)</td>
			<td>Fisher Scientific</td>
			<td></td>
			<td>To make 70% ethanol for cleaning skin.</td>
		</tr>
		<tr>
			<td>Hindlimb Suspension Cages</td>
			<td>Custom Made</td>
			<td>N/A</td>
			<td>These custom made cages will be highlighted in the video recordings of the MS.</td>
		</tr>
		<tr>
			<td>Optixcare Eye Lube</td>
			<td>Optixcare</td>
			<td></td>
			<td>Eye lube for protection during survival surgery.</td>
		</tr>
		<tr>
			<td>Scalpel blades #11</td>
			<td>Fine Science</td>
			<td></td>
			<td>Blade used to make incisions on skin and muscle.</td>
		</tr>
		<tr>
			<td>Skin-Trac</td>
			<td>Zimmer</td>
			<td>736579</td>
			<td>Foam tape for fixing the tail to the suspension apparatus.</td>
		</tr>
		<tr>
			<td>SomnoSuite Low-Flow Digital Vaporizer</td>
			<td>Kent Scientific Corporation</td>
			<td>SS-01</td>
			<td>Vaporizer for Isoflurane Anesthesia</td>
		</tr>
		<tr>
			<td>Tissue bath apparatus&#160;</td>
			<td>Aurora Scientific</td>
			<td>Model 800A, Dual Mode Muscle Lever 300C</td>
			<td></td>
		</tr>
	</tbody>
</table>

a preclinical model of sepsis-induced myopathy with disuse in mice

Sepsis is a life-threatening condition due to an overactive immune response that adversely affects multiple organ systems, resulting in a major burden to health systems worldwide1. More recently, the in-hospital mortality linked to sepsis has decreased due to improved intensity care unit (ICU) management1,2. However, approximately 75% of the patients surviving the initial septic insult develop skeletal muscle atrophy (e.g., reductions in cross-sectional area) and weakness (e.g., reductions in force production capacity)3,4. This phenomenon has been characterized as sepsis-induced myopathy, highly linked to impaired physical activity and lack of independence to perform daily living tasks, leading to re-hospitalization and mortality within five years of the initial episode5.
Due to an aggressive and generalized infection, septic patients are exposed to prolonged periods of bed rest while recovering in the ICU. In this context, the skeletal muscle undergoes severe disuse, which likely exacerbates muscle atrophy and weakness3,4. Currently, no treatment has effectively addressed sepsis-induced myopathy. The available preclinical models designed to address the myopathy have used cecal ligation and puncture (CLP)6, cecal slurry7, or injection of purified lipopolysaccharide (LPS), which is a component of the cellular wall in gram-negative bacteria8. Although these models succeed in delivering infection, they do not properly reproduce the muscle disuse observed in septic hosts beyond a natural reduction in physical activity observed in septic animals9. 
The main objective of this study is to provide a detailed description of how to properly execute the model of sepsis-induced myopathy with disuse in mice. We demonstrate the feasibility of combining CLP as a model of sepsis with hindlimb suspension (HLS) as a model of disuse to study sepsis-induced myopathy in mice3. Furthermore, representative results of muscle mechanics and typical morphological changes in response to the model are also provided.

<meta charset="utf8"></head><body>저자 스포트라이트: 패혈증 유발 근병증 및 단일 세포 대사 분석을 위한 고급 통합 모델

A Preclinical model of Sepsis-Induced Myopathy with Disuse in mouse(마우스에서 사용하지 않는 패혈증 유발 근병증의 전임상 모델)

Our research focuses on understanding the mechanisms behind sepsis induced myopathy. Current models overlook the disuse component, which is crucial for bad reading septic patients. We aim to integrate this aspect for a more accurate disease model.We have significantly advanced the field by combining cecal ligation and puncture model with high limb suspension. This integrated approach allow us to study the systematic effect of sepsis and the impact of muscle disuse on sepsis induced myopathy, providing a more realistic model for understanding the disease progression and potential interventions. Our protocol combines cecal ligation and puncture with high limb suspension to replicate the clinical scenario of human sepsis more accurately in mice.By simulating muscle atrophy and weakness seen in septic patients, our approach provides a comprehensive model for studying the interplay between sepsis and muscle function, allowing for more targeted investigations into therapeutic interventions.

Sepsis is a major cause of in-hospital deaths. Improvements in treatment result in a greater number of sepsis survivors. Approximately 75% of the ...

a-preclinical-model-of-sepsis-induced-myopathy-with-disuse-in-mice

Research

JoVE Journal

Medicine

A Preclinical Model of Sepsis-Induced Myopathy with Disuse in Mice

375 조회수.  University of Florida. 우리의 연구는 패혈증 유발 근육병증의 기전을 이해하는 데 중점을 둡니다. 현재 모델은 판독이 나쁜 패혈증 환자에게 중요한 불용 구성 요소를 간과합니다. 우리는 보다 정확한 질병 모델을 위해 이 측면을 통합하는 것을 목표로 합니다.우리는 cecal ligation 및 puncture model을 high limb suspension과 결합하여 이 분야를 크게 발전시켰습니다. 이 통합 접...

비디오: 저자 스포트라이트: 패혈증 유발 근병증 및 단일 세포 대사 분석을 위한 고급 통합 모델

Sepsis is a major cause of in-hospital deaths. Improvements in treatment result in a greater number of sepsis survivors. Approximately 75% of the survivors develop muscle weakness and atrophy, increasing the incidence of hospital readmissions and mortality. However, the available preclinical models of sepsis do not address skeletal muscle disuse, a key component for the development of sepsis-induced myopathy. Our objective in this protocol is to provide a step-by-step guideline for a mouse model that reproduces the clinical setting experienced by a bedridden septic patient. Male C57Bl/6 mice were used to develop this model. Mice underwent cecal ligation and puncture (CLP) to induce sepsis. Four days post-CLP, mice were subjected to hindlimb suspension (HLS) for seven days. Results were compared with sham-matched surgeries and/or animals with normal ambulation (NA). Muscles were dissected for in vitro muscle mechanics and morphological assessments. The model results in marked muscle atrophy and weakness, a similar phenotype observed in septic patients. The model represents a platform for testing potential therapeutic strategies for the mitigation of sepsis-induced myopathy.

This murine model combines a septic insult with hindlimb muscle disuse to recapitulate the bedridden feature of the typical septic patient. The model represents a significant departure from previous models to study muscle dysfunction in sepsis and is a reproducible approach to addressing therapeutic strategies to treat this condition.

연구

의학

Combining Cecal Ligation and Puncture (CLP) with Hindlimb Suspension (HLS) to Study Sepsis-Induced Myopathy in Mice

To begin, transfer the prepared anesthetized mouse to the surgical area and isolate the surgical site with an adhesive drape. Under deep anesthesia, use a scalpel blade to make a ventral midline incision in the skin. Next, use scissors to separate the skin from the muscle layer.With the help of a scalpel blade, make a smaller incision in the muscle layer. Once the bowels are visualized, use blunt forceps to locate the cecum and exteriorize it. Then ligate the cecum with a sterile 5/0 polyglactin absorbable suture.Using a 27-gauge needle, perforate the secum through and through allowing the fecal content to leak. Gently squeeze the secum to externalize the fecal content with caution. Afterward, relocate the secum into the abdominal cavity and close the muscle layer with a sterile 5/0 absorbable suture.Then use a 5/0 nylon non-absorbable suture to close the skin. After that, provide sterile saline via subcutaneous injection in the back of the animal. After four days of recovery from CLP, or sham surgery, use foam tape to attach the anesthetized mouse tail to a short metal chain.Then attach the metal chain to a hook connected to a crossbar along the center of the cage. Affix a second small bar that can move along the crossbar to allow the animal greater movement ability. Then adjust the height of the suspended limbs to prevent the paws from contacting the chow pellets.Ensure that the animals can move via their forelimbs using the metal grid on the cage floor. CLP animals demonstrated increased immune cell populations in their peritoneal lavage compared to sham animals. CLP and HLS lead to approximately 11.8%of body mass loss.A marked decrease in soleus mass was observed when HLS was superimposed with CLP, indicating that smaller fibers comprise a major part of the muscle section. Compared to the control animals, CLP-HLS caused significant impairment in both the absolute and peak forces of the soleus.