Role of Autophagy and Apoptosis in Wound Tissue of Deep Second‐degree Burn in Rats
Objectives The pathogenesis of burn wound progression is poorly understood. Contributing factors include continuous loss of blood perfusion, excessive inflammation, and elevated apoptosis levels in wound tissue. Macroautophagy (here referred to simply as “autophagy”) is associated with many chronic...
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| Veröffentlicht in: | Academic emergency medicine 2014-04, Vol.21 (4), p.383-391 |
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| creator | Xiao, Mengjing Li, Ligen Li, Chenxi Zhang, Peirong Hu, Quan Ma, Li Zhang, Haijun Olson, James E. |
| description | Objectives
The pathogenesis of burn wound progression is poorly understood. Contributing factors include continuous loss of blood perfusion, excessive inflammation, and elevated apoptosis levels in wound tissue. Macroautophagy (here referred to simply as “autophagy”) is associated with many chronic diseases. The authors hypothesized that autophagy is involved in burn wound progression in a rat model of deep second‐degree burn.
Methods
Deep second‐degree burns were modeled using a brass rod heated to 100°C applied for 6 seconds to the back skin of Wistar rats. Full‐thickness biopsies were obtained from burned and nonburned controls at several times postburn. Western blotting and immunohistochemical (IHC) staining determined expression of the autophagy markers Light Chain 3 (LC3) and beclin‐1. Apoptosis was determined by terminal‐deoxynucleoitidyl transferase mediated nick end labeling (TUNEL) assay and laser Doppler flowmetry (LDF)‐measured tissue perfusion. Myeloperoxidase (MPO) activity assay measured inflammation. Hematoxylin and eosin (H&E) and Masson's trichrome staining–determined pathology and wound depth.
Results
The LC3 and beclin‐1 protein level in burn wounds decreased to one‐fourth of normal levels (p |
| doi_str_mv | 10.1111/acem.12352 |
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| fullrecord | <record><control><sourceid>proquest_pubme</sourceid><recordid>TN_cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_4114170</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>1526729640</sourcerecordid><originalsourceid>FETCH-LOGICAL-c3632-3a9583a41f1276a236e0f5b18d084794485e18df870694c51ce5799c736accac3</originalsourceid><addsrcrecordid>eNp9kctO3DAUhi0EKpd20weoIrFBSAEfX5MN0jBQQKKqNJ2KpWWckyEoE4c4Ac2OR-gz9knqYQbUsqg3vn369B_9hHwGegRxHVuH8yNgXLINsgNS8pRpYJvxTFWeKqn4NtkN4Z5SKnWuP5BtJjSngtIdMp34GhNfJqOh9-2dnS0S2xTJqPVt70MVkqpJbvwQn6ZVCMMLeobYJj_Q-ab4_fyrwFmHmJwOXbOEJ7YPH8lWaeuAn9b7Hvn59Xw6vkyvv19cjUfXqeOKs5TbXGbcCiiBaWUZV0hLeQtZQTOhcyEyifFSZjrOIZwEhzF_7jRX1jnr-B45WXnb4XaOhcOm72xt2q6a225hvK3Mvz9NdWdm_tEIAAGaRsHBWtD5hwFDb-ZVcFjXtkE_BAOSKc1yJZbo_jv03seR43iRgjwagYlIHa4o1_kQOizfwgA1y7LMsizzUlaEv_wd_w19bScCsAKeqhoX_1GZ0fj820r6BwYYnlw</addsrcrecordid><sourcetype>Open Access Repository</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>1519411124</pqid></control><display><type>article</type><title>Role of Autophagy and Apoptosis in Wound Tissue of Deep Second‐degree Burn in Rats</title><source>MEDLINE</source><source>Access via Wiley Online Library</source><source>EZB-FREE-00999 freely available EZB journals</source><source>Wiley Online Library (Open Access Collection)</source><creator>Xiao, Mengjing ; Li, Ligen ; Li, Chenxi ; Zhang, Peirong ; Hu, Quan ; Ma, Li ; Zhang, Haijun ; Olson, James E.</creator><contributor>Olson, James E.</contributor><creatorcontrib>Xiao, Mengjing ; Li, Ligen ; Li, Chenxi ; Zhang, Peirong ; Hu, Quan ; Ma, Li ; Zhang, Haijun ; Olson, James E. ; Olson, James E.</creatorcontrib><description>Objectives
The pathogenesis of burn wound progression is poorly understood. Contributing factors include continuous loss of blood perfusion, excessive inflammation, and elevated apoptosis levels in wound tissue. Macroautophagy (here referred to simply as “autophagy”) is associated with many chronic diseases. The authors hypothesized that autophagy is involved in burn wound progression in a rat model of deep second‐degree burn.
Methods
Deep second‐degree burns were modeled using a brass rod heated to 100°C applied for 6 seconds to the back skin of Wistar rats. Full‐thickness biopsies were obtained from burned and nonburned controls at several times postburn. Western blotting and immunohistochemical (IHC) staining determined expression of the autophagy markers Light Chain 3 (LC3) and beclin‐1. Apoptosis was determined by terminal‐deoxynucleoitidyl transferase mediated nick end labeling (TUNEL) assay and laser Doppler flowmetry (LDF)‐measured tissue perfusion. Myeloperoxidase (MPO) activity assay measured inflammation. Hematoxylin and eosin (H&E) and Masson's trichrome staining–determined pathology and wound depth.
Results
The LC3 and beclin‐1 protein level in burn wounds decreased to one‐fourth of normal levels (p < 0.01) over 24 hours and then began to increase but still did not reach their normal level. TUNEL‐positive cells in burn wounds were 3.7‐fold (p < 0.01) elevated over 48 hours and then decreased slightly, yet still remained higher than in normal skin. The burn wound progressed in depth over 72 hours. In addition, significant decrease in LDF values and upregulation of MPO activity were observed. Enhanced LC3‐positive cells were observed in the deep dermal layer of burn wounds as shown by IHC staining.
Conclusions
A reduction in autophagy and blood flow and an increase in apoptosis and inflammation were observed in burn wounds early during the course of burn injury progression. This suggests that autophagy, complemented by apoptosis, play important roles in burn progression. Enhanced autophagy in the deep dermis may be a prosurvival mechanism against ischemia and inflammation after burn injury.
Resumen
Objetivos
La patogénesis de la progresión en la lesión por quemadura es poco conocida. Los factores contribuyentes incluyen la pérdida continua de perfusión sanguínea, la excesiva inflamación y los marcadores de apoptosis elevados en el tejido lesionado. La macroautofagia (aquí referida simplemente como “autofagia”) se asocia con muchas enfermedades crónicas. Se generó la hipótesis que la autofagia está implicada en la progresión de la lesión por quemadura en un modelo de quemadura de segundo grado en ratas.
Metodología
Las quemaduras de segundo grado se simularon usando un varilla de metal calentada hasta 100°C y se aplicó durante 6 segundos en la piel del dorso de las ratas Wistar. Se obtuvieron biopsias de todo el grosor de los quemados y de los controles no quemados en diversas ocasiones tras la quemadura. Se determinó la expresión de marcadores de autofagia Light Chain 3 (LC3), y Beclin‐1 mediante Wetern blot y tinción inmunohistoquímica. La apoptosis se determinó por técnica TUNEL (TUNEL assay) y la flujometría Doppler láser midió la perfusión del tejido. La técnica de actividad mieloperoxidasa midió la inflamación. Las tinciones con hematoxilina y eosina, y con tricrómico de Masson determinaron la patología y profundidad de la lesión.
Resultados
El valor de proteínas Beclin‐1 y LC3 en lesiones por quemadura disminuyó a un cuarto del valor normal (p < 0,01) a las 24 horas, y después empezó a incrementarse pero no alcanzó sus valores normales. Las células positivas TUNEL en las lesiones por quemadura fueron 3,7 veces (p < 0,01) más elevadas a las 48 horas, y después disminuyeron ligeramente, permaneciendo todavía más altas que en la piel normal. La lesión por quemadura progresó en profundidad durante 72 horas. Además, se observaron un descenso significativo en los valores de flujometría Doppler láser y un aumento de la actividad mieloperoxidasa. Se observaron mediante células positivas LC3 aumentadas en la dermis profunda de las lesions por quemadure la tíncion inmunohistoquímica.
Conclusiones
Se observaron una reducción de la autofagia y del flujo sanguíneo y un incremento en la apoptosis y la inflamación en las heridas por quemadura al inicio del curso de la progresión de la lesión. Esto indica que la autofagia, junto con la apoptosis, juegan papeles importantes en la progresión de la quemadura. El incremento de la autofagia en la dermis profunda puede ser un mecanismo a favor de la supervivencia y que actúa en contra de la isquemia y la inflamación tras la lesión por quemadura.</description><identifier>ISSN: 1069-6563</identifier><identifier>EISSN: 1553-2712</identifier><identifier>DOI: 10.1111/acem.12352</identifier><identifier>PMID: 24730400</identifier><language>eng</language><publisher>United States: Wiley Subscription Services, Inc</publisher><subject>Animals ; Apoptosis ; Autophagy ; Biomarkers - metabolism ; Blotting, Western ; Burns ; Burns - metabolism ; Burns - pathology ; Burns - physiopathology ; Disease Progression ; Immunohistochemistry ; In Situ Nick-End Labeling ; Male ; Original Contribution ; Original Research Contributions ; Random Allocation ; Rats ; Rats, Wistar ; Rodents ; Wound healing</subject><ispartof>Academic emergency medicine, 2014-04, Vol.21 (4), p.383-391</ispartof><rights>2014 The Authors. published by Wiley Periodicals, Inc. on behalf of Society for Academic Emergency Medicine (SAEM).</rights><rights>2014 The Authors. Academic Emergency Medicine published by Wiley Periodicals, Inc. on behalf of Society for Academic Emergency Medicine (SAEM).</rights><rights>Copyright Hanley & Belfus, Inc. Apr 2014</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c3632-3a9583a41f1276a236e0f5b18d084794485e18df870694c51ce5799c736accac3</citedby><cites>FETCH-LOGICAL-c3632-3a9583a41f1276a236e0f5b18d084794485e18df870694c51ce5799c736accac3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>230,315,781,785,886,1418,1434,27929,27930</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/24730400$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Olson, James E.</contributor><creatorcontrib>Xiao, Mengjing</creatorcontrib><creatorcontrib>Li, Ligen</creatorcontrib><creatorcontrib>Li, Chenxi</creatorcontrib><creatorcontrib>Zhang, Peirong</creatorcontrib><creatorcontrib>Hu, Quan</creatorcontrib><creatorcontrib>Ma, Li</creatorcontrib><creatorcontrib>Zhang, Haijun</creatorcontrib><creatorcontrib>Olson, James E.</creatorcontrib><title>Role of Autophagy and Apoptosis in Wound Tissue of Deep Second‐degree Burn in Rats</title><title>Academic emergency medicine</title><addtitle>Acad Emerg Med</addtitle><description>Objectives
The pathogenesis of burn wound progression is poorly understood. Contributing factors include continuous loss of blood perfusion, excessive inflammation, and elevated apoptosis levels in wound tissue. Macroautophagy (here referred to simply as “autophagy”) is associated with many chronic diseases. The authors hypothesized that autophagy is involved in burn wound progression in a rat model of deep second‐degree burn.
Methods
Deep second‐degree burns were modeled using a brass rod heated to 100°C applied for 6 seconds to the back skin of Wistar rats. Full‐thickness biopsies were obtained from burned and nonburned controls at several times postburn. Western blotting and immunohistochemical (IHC) staining determined expression of the autophagy markers Light Chain 3 (LC3) and beclin‐1. Apoptosis was determined by terminal‐deoxynucleoitidyl transferase mediated nick end labeling (TUNEL) assay and laser Doppler flowmetry (LDF)‐measured tissue perfusion. Myeloperoxidase (MPO) activity assay measured inflammation. Hematoxylin and eosin (H&E) and Masson's trichrome staining–determined pathology and wound depth.
Results
The LC3 and beclin‐1 protein level in burn wounds decreased to one‐fourth of normal levels (p < 0.01) over 24 hours and then began to increase but still did not reach their normal level. TUNEL‐positive cells in burn wounds were 3.7‐fold (p < 0.01) elevated over 48 hours and then decreased slightly, yet still remained higher than in normal skin. The burn wound progressed in depth over 72 hours. In addition, significant decrease in LDF values and upregulation of MPO activity were observed. Enhanced LC3‐positive cells were observed in the deep dermal layer of burn wounds as shown by IHC staining.
Conclusions
A reduction in autophagy and blood flow and an increase in apoptosis and inflammation were observed in burn wounds early during the course of burn injury progression. This suggests that autophagy, complemented by apoptosis, play important roles in burn progression. Enhanced autophagy in the deep dermis may be a prosurvival mechanism against ischemia and inflammation after burn injury.
Resumen
Objetivos
La patogénesis de la progresión en la lesión por quemadura es poco conocida. Los factores contribuyentes incluyen la pérdida continua de perfusión sanguínea, la excesiva inflamación y los marcadores de apoptosis elevados en el tejido lesionado. La macroautofagia (aquí referida simplemente como “autofagia”) se asocia con muchas enfermedades crónicas. Se generó la hipótesis que la autofagia está implicada en la progresión de la lesión por quemadura en un modelo de quemadura de segundo grado en ratas.
Metodología
Las quemaduras de segundo grado se simularon usando un varilla de metal calentada hasta 100°C y se aplicó durante 6 segundos en la piel del dorso de las ratas Wistar. Se obtuvieron biopsias de todo el grosor de los quemados y de los controles no quemados en diversas ocasiones tras la quemadura. Se determinó la expresión de marcadores de autofagia Light Chain 3 (LC3), y Beclin‐1 mediante Wetern blot y tinción inmunohistoquímica. La apoptosis se determinó por técnica TUNEL (TUNEL assay) y la flujometría Doppler láser midió la perfusión del tejido. La técnica de actividad mieloperoxidasa midió la inflamación. Las tinciones con hematoxilina y eosina, y con tricrómico de Masson determinaron la patología y profundidad de la lesión.
Resultados
El valor de proteínas Beclin‐1 y LC3 en lesiones por quemadura disminuyó a un cuarto del valor normal (p < 0,01) a las 24 horas, y después empezó a incrementarse pero no alcanzó sus valores normales. Las células positivas TUNEL en las lesiones por quemadura fueron 3,7 veces (p < 0,01) más elevadas a las 48 horas, y después disminuyeron ligeramente, permaneciendo todavía más altas que en la piel normal. La lesión por quemadura progresó en profundidad durante 72 horas. Además, se observaron un descenso significativo en los valores de flujometría Doppler láser y un aumento de la actividad mieloperoxidasa. Se observaron mediante células positivas LC3 aumentadas en la dermis profunda de las lesions por quemadure la tíncion inmunohistoquímica.
Conclusiones
Se observaron una reducción de la autofagia y del flujo sanguíneo y un incremento en la apoptosis y la inflamación en las heridas por quemadura al inicio del curso de la progresión de la lesión. Esto indica que la autofagia, junto con la apoptosis, juegan papeles importantes en la progresión de la quemadura. El incremento de la autofagia en la dermis profunda puede ser un mecanismo a favor de la supervivencia y que actúa en contra de la isquemia y la inflamación tras la lesión por quemadura.</description><subject>Animals</subject><subject>Apoptosis</subject><subject>Autophagy</subject><subject>Biomarkers - metabolism</subject><subject>Blotting, Western</subject><subject>Burns</subject><subject>Burns - metabolism</subject><subject>Burns - pathology</subject><subject>Burns - physiopathology</subject><subject>Disease Progression</subject><subject>Immunohistochemistry</subject><subject>In Situ Nick-End Labeling</subject><subject>Male</subject><subject>Original Contribution</subject><subject>Original Research Contributions</subject><subject>Random Allocation</subject><subject>Rats</subject><subject>Rats, Wistar</subject><subject>Rodents</subject><subject>Wound healing</subject><issn>1069-6563</issn><issn>1553-2712</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2014</creationdate><recordtype>article</recordtype><sourceid>24P</sourceid><sourceid>WIN</sourceid><sourceid>EIF</sourceid><recordid>eNp9kctO3DAUhi0EKpd20weoIrFBSAEfX5MN0jBQQKKqNJ2KpWWckyEoE4c4Ac2OR-gz9knqYQbUsqg3vn369B_9hHwGegRxHVuH8yNgXLINsgNS8pRpYJvxTFWeKqn4NtkN4Z5SKnWuP5BtJjSngtIdMp34GhNfJqOh9-2dnS0S2xTJqPVt70MVkqpJbvwQn6ZVCMMLeobYJj_Q-ab4_fyrwFmHmJwOXbOEJ7YPH8lWaeuAn9b7Hvn59Xw6vkyvv19cjUfXqeOKs5TbXGbcCiiBaWUZV0hLeQtZQTOhcyEyifFSZjrOIZwEhzF_7jRX1jnr-B45WXnb4XaOhcOm72xt2q6a225hvK3Mvz9NdWdm_tEIAAGaRsHBWtD5hwFDb-ZVcFjXtkE_BAOSKc1yJZbo_jv03seR43iRgjwagYlIHa4o1_kQOizfwgA1y7LMsizzUlaEv_wd_w19bScCsAKeqhoX_1GZ0fj820r6BwYYnlw</recordid><startdate>201404</startdate><enddate>201404</enddate><creator>Xiao, Mengjing</creator><creator>Li, Ligen</creator><creator>Li, Chenxi</creator><creator>Zhang, Peirong</creator><creator>Hu, Quan</creator><creator>Ma, Li</creator><creator>Zhang, Haijun</creator><creator>Olson, James E.</creator><general>Wiley Subscription Services, Inc</general><general>Wiley</general><scope>24P</scope><scope>WIN</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>K9.</scope><scope>NAPCQ</scope><scope>U9A</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>201404</creationdate><title>Role of Autophagy and Apoptosis in Wound Tissue of Deep Second‐degree Burn in Rats</title><author>Xiao, Mengjing ; Li, Ligen ; Li, Chenxi ; Zhang, Peirong ; Hu, Quan ; Ma, Li ; Zhang, Haijun ; Olson, James E.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c3632-3a9583a41f1276a236e0f5b18d084794485e18df870694c51ce5799c736accac3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2014</creationdate><topic>Animals</topic><topic>Apoptosis</topic><topic>Autophagy</topic><topic>Biomarkers - metabolism</topic><topic>Blotting, Western</topic><topic>Burns</topic><topic>Burns - metabolism</topic><topic>Burns - pathology</topic><topic>Burns - physiopathology</topic><topic>Disease Progression</topic><topic>Immunohistochemistry</topic><topic>In Situ Nick-End Labeling</topic><topic>Male</topic><topic>Original Contribution</topic><topic>Original Research Contributions</topic><topic>Random Allocation</topic><topic>Rats</topic><topic>Rats, Wistar</topic><topic>Rodents</topic><topic>Wound healing</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Xiao, Mengjing</creatorcontrib><creatorcontrib>Li, Ligen</creatorcontrib><creatorcontrib>Li, Chenxi</creatorcontrib><creatorcontrib>Zhang, Peirong</creatorcontrib><creatorcontrib>Hu, Quan</creatorcontrib><creatorcontrib>Ma, Li</creatorcontrib><creatorcontrib>Zhang, Haijun</creatorcontrib><creatorcontrib>Olson, James E.</creatorcontrib><collection>Wiley Online Library (Open Access Collection)</collection><collection>Wiley Online Library (Open Access Collection)</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Nursing & Allied Health Premium</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Academic emergency medicine</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Xiao, Mengjing</au><au>Li, Ligen</au><au>Li, Chenxi</au><au>Zhang, Peirong</au><au>Hu, Quan</au><au>Ma, Li</au><au>Zhang, Haijun</au><au>Olson, James E.</au><au>Olson, James E.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Role of Autophagy and Apoptosis in Wound Tissue of Deep Second‐degree Burn in Rats</atitle><jtitle>Academic emergency medicine</jtitle><addtitle>Acad Emerg Med</addtitle><date>2014-04</date><risdate>2014</risdate><volume>21</volume><issue>4</issue><spage>383</spage><epage>391</epage><pages>383-391</pages><issn>1069-6563</issn><eissn>1553-2712</eissn><abstract>Objectives
The pathogenesis of burn wound progression is poorly understood. Contributing factors include continuous loss of blood perfusion, excessive inflammation, and elevated apoptosis levels in wound tissue. Macroautophagy (here referred to simply as “autophagy”) is associated with many chronic diseases. The authors hypothesized that autophagy is involved in burn wound progression in a rat model of deep second‐degree burn.
Methods
Deep second‐degree burns were modeled using a brass rod heated to 100°C applied for 6 seconds to the back skin of Wistar rats. Full‐thickness biopsies were obtained from burned and nonburned controls at several times postburn. Western blotting and immunohistochemical (IHC) staining determined expression of the autophagy markers Light Chain 3 (LC3) and beclin‐1. Apoptosis was determined by terminal‐deoxynucleoitidyl transferase mediated nick end labeling (TUNEL) assay and laser Doppler flowmetry (LDF)‐measured tissue perfusion. Myeloperoxidase (MPO) activity assay measured inflammation. Hematoxylin and eosin (H&E) and Masson's trichrome staining–determined pathology and wound depth.
Results
The LC3 and beclin‐1 protein level in burn wounds decreased to one‐fourth of normal levels (p < 0.01) over 24 hours and then began to increase but still did not reach their normal level. TUNEL‐positive cells in burn wounds were 3.7‐fold (p < 0.01) elevated over 48 hours and then decreased slightly, yet still remained higher than in normal skin. The burn wound progressed in depth over 72 hours. In addition, significant decrease in LDF values and upregulation of MPO activity were observed. Enhanced LC3‐positive cells were observed in the deep dermal layer of burn wounds as shown by IHC staining.
Conclusions
A reduction in autophagy and blood flow and an increase in apoptosis and inflammation were observed in burn wounds early during the course of burn injury progression. This suggests that autophagy, complemented by apoptosis, play important roles in burn progression. Enhanced autophagy in the deep dermis may be a prosurvival mechanism against ischemia and inflammation after burn injury.
Resumen
Objetivos
La patogénesis de la progresión en la lesión por quemadura es poco conocida. Los factores contribuyentes incluyen la pérdida continua de perfusión sanguínea, la excesiva inflamación y los marcadores de apoptosis elevados en el tejido lesionado. La macroautofagia (aquí referida simplemente como “autofagia”) se asocia con muchas enfermedades crónicas. Se generó la hipótesis que la autofagia está implicada en la progresión de la lesión por quemadura en un modelo de quemadura de segundo grado en ratas.
Metodología
Las quemaduras de segundo grado se simularon usando un varilla de metal calentada hasta 100°C y se aplicó durante 6 segundos en la piel del dorso de las ratas Wistar. Se obtuvieron biopsias de todo el grosor de los quemados y de los controles no quemados en diversas ocasiones tras la quemadura. Se determinó la expresión de marcadores de autofagia Light Chain 3 (LC3), y Beclin‐1 mediante Wetern blot y tinción inmunohistoquímica. La apoptosis se determinó por técnica TUNEL (TUNEL assay) y la flujometría Doppler láser midió la perfusión del tejido. La técnica de actividad mieloperoxidasa midió la inflamación. Las tinciones con hematoxilina y eosina, y con tricrómico de Masson determinaron la patología y profundidad de la lesión.
Resultados
El valor de proteínas Beclin‐1 y LC3 en lesiones por quemadura disminuyó a un cuarto del valor normal (p < 0,01) a las 24 horas, y después empezó a incrementarse pero no alcanzó sus valores normales. Las células positivas TUNEL en las lesiones por quemadura fueron 3,7 veces (p < 0,01) más elevadas a las 48 horas, y después disminuyeron ligeramente, permaneciendo todavía más altas que en la piel normal. La lesión por quemadura progresó en profundidad durante 72 horas. Además, se observaron un descenso significativo en los valores de flujometría Doppler láser y un aumento de la actividad mieloperoxidasa. Se observaron mediante células positivas LC3 aumentadas en la dermis profunda de las lesions por quemadure la tíncion inmunohistoquímica.
Conclusiones
Se observaron una reducción de la autofagia y del flujo sanguíneo y un incremento en la apoptosis y la inflamación en las heridas por quemadura al inicio del curso de la progresión de la lesión. Esto indica que la autofagia, junto con la apoptosis, juegan papeles importantes en la progresión de la quemadura. El incremento de la autofagia en la dermis profunda puede ser un mecanismo a favor de la supervivencia y que actúa en contra de la isquemia y la inflamación tras la lesión por quemadura.</abstract><cop>United States</cop><pub>Wiley Subscription Services, Inc</pub><pmid>24730400</pmid><doi>10.1111/acem.12352</doi><tpages>9</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | Academic emergency medicine, 2014-04, Vol.21 (4), p.383-391 |
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| source | MEDLINE; Access via Wiley Online Library; EZB-FREE-00999 freely available EZB journals; Wiley Online Library (Open Access Collection) |
| subjects | Animals Apoptosis Autophagy Biomarkers - metabolism Blotting, Western Burns Burns - metabolism Burns - pathology Burns - physiopathology Disease Progression Immunohistochemistry In Situ Nick-End Labeling Male Original Contribution Original Research Contributions Random Allocation Rats Rats, Wistar Rodents Wound healing |
| title | Role of Autophagy and Apoptosis in Wound Tissue of Deep Second‐degree Burn in Rats |
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