Evaluation of macroautophagy in an experimental model similar to lupus nephritis
Systemic lupus erythematosus (SLE) is a complex and frequent autoimmune disease, presenting with lupus nephritis (LN) in 30-60% of patients. Macroautophagy, a mechanism for the degradation of intracellular contents, is deregulated in numerous metabolic, neurodegenerative, cancer and autoimmune disea...
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| Formato: | Artículo revista |
| Lenguaje: | Español |
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Universidad Nacional Córdoba. Facultad de Ciencias Médicas. Secretaria de Ciencia y Tecnología
2023
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| Acceso en línea: | https://revistas.unc.edu.ar/index.php/med/article/view/42771 |
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I10-R327-article-42771 |
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ojs |
| institution |
Universidad Nacional de Córdoba |
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I-10 |
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R-327 |
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Revista de la Facultad de Ciencias Médicas de Córdoba |
| language |
Español |
| format |
Artículo revista |
| topic |
LUPUS ERITEMATOSO SISTÉMICO NEFRITIS LÚPICA BECLINA 1 LC3 MICROSCOPIA ELECTRÓNICA DE TRANSMISIÓN |
| spellingShingle |
LUPUS ERITEMATOSO SISTÉMICO NEFRITIS LÚPICA BECLINA 1 LC3 MICROSCOPIA ELECTRÓNICA DE TRANSMISIÓN Faure, EE Gutiérrez, S Mukdsi, JH Evaluation of macroautophagy in an experimental model similar to lupus nephritis |
| topic_facet |
LUPUS ERITEMATOSO SISTÉMICO NEFRITIS LÚPICA BECLINA 1 LC3 MICROSCOPIA ELECTRÓNICA DE TRANSMISIÓN |
| author |
Faure, EE Gutiérrez, S Mukdsi, JH |
| author_facet |
Faure, EE Gutiérrez, S Mukdsi, JH |
| author_sort |
Faure, EE |
| title |
Evaluation of macroautophagy in an experimental model similar to lupus nephritis |
| title_short |
Evaluation of macroautophagy in an experimental model similar to lupus nephritis |
| title_full |
Evaluation of macroautophagy in an experimental model similar to lupus nephritis |
| title_fullStr |
Evaluation of macroautophagy in an experimental model similar to lupus nephritis |
| title_full_unstemmed |
Evaluation of macroautophagy in an experimental model similar to lupus nephritis |
| title_sort |
evaluation of macroautophagy in an experimental model similar to lupus nephritis |
| description |
Systemic lupus erythematosus (SLE) is a complex and frequent autoimmune disease, presenting with lupus nephritis (LN) in 30-60% of patients. Macroautophagy, a mechanism for the degradation of intracellular contents, is deregulated in numerous metabolic, neurodegenerative, cancer and autoimmune diseases. Although autophagy has been shown to be involved in its pathogenesis, to what extent it is present in target organs remains uncertain. We set out to analyze and describe the presence of macroautophagy in an NL-like experimental model.
Male and female knockout mice for Galectin-3 of 8 months of age (n: 6 for each sex) were used, in which ANAs and renal functional compromise (proteinuria and increased creatinine) have been described, being the controls (C) mice of both sexes of the B6 strain (n: 6 per sex). The methodologies used for the evaluation of macroautophagy were: transmission electron microscopy (4% Karnovsky fixative, embedding in epoxy resins, contrast with heavy metals, observation in a Zeiss Leo 906-E electron microscope) and immunohistochemistry in paraffin for Beclina-1 and LC3, determining the number of immunopositive cells per tubule, counting between 710 and 790 tubules per control and experimental group. Statistics: mean and SD, Mann-Whitney test (two groups quantitative variable, unpaired, without normal distribution). p<0.05. Protocol approved by CICUAL-University of San Martín.
Histopathological evaluation demonstrated, in both male and female NL-like mice, immune complex-mediated mesangial proliferative glomerulonephritis and chronic tubulointerstitial nephritis. The ultrastructural analysis of the experimental groups, unlike the control group and regardless of sex, demonstrated vesicular elements in tubules compatible with macroautophagic vesicles. Both Beclin-1 and LC3 were immunodetected in the renal tubular epithelium, with a significantly higher number of immunopositive cells in the NL-like experimental model compared to the control (example Beclin 1 male C:5±3; NL-like:20±7 *, female C:7±2, similar-NL:25±4** *p<0.05 vs C and **p<0.05 vs C).
We can conclude that in the NL-like model the macroautophagy mechanism is increased compared to control animals, possibly linked to the proteinuric state of this strain, and may also contribute to the chronic interstitial damage described. |
| publisher |
Universidad Nacional Córdoba. Facultad de Ciencias Médicas. Secretaria de Ciencia y Tecnología |
| publishDate |
2023 |
| url |
https://revistas.unc.edu.ar/index.php/med/article/view/42771 |
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AT faureee evaluationofmacroautophagyinanexperimentalmodelsimilartolupusnephritis AT gutierrezs evaluationofmacroautophagyinanexperimentalmodelsimilartolupusnephritis AT mukdsijh evaluationofmacroautophagyinanexperimentalmodelsimilartolupusnephritis AT faureee evaluaciondelamacroautofagiaenunmodeloexperimentalsimilnefritislupica AT gutierrezs evaluaciondelamacroautofagiaenunmodeloexperimentalsimilnefritislupica AT mukdsijh evaluaciondelamacroautofagiaenunmodeloexperimentalsimilnefritislupica |
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2024-09-03T21:04:57Z |
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2024-09-03T21:04:57Z |
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I10-R327-article-427712023-10-19T21:19:21Z Evaluation of macroautophagy in an experimental model similar to lupus nephritis Evaluación de la macroautofagia en un modelo experimental símil nefritis lúpica Faure, EE Gutiérrez, S Mukdsi, JH LUPUS ERITEMATOSO SISTÉMICO NEFRITIS LÚPICA BECLINA 1 LC3 MICROSCOPIA ELECTRÓNICA DE TRANSMISIÓN Systemic lupus erythematosus (SLE) is a complex and frequent autoimmune disease, presenting with lupus nephritis (LN) in 30-60% of patients. Macroautophagy, a mechanism for the degradation of intracellular contents, is deregulated in numerous metabolic, neurodegenerative, cancer and autoimmune diseases. Although autophagy has been shown to be involved in its pathogenesis, to what extent it is present in target organs remains uncertain. We set out to analyze and describe the presence of macroautophagy in an NL-like experimental model. Male and female knockout mice for Galectin-3 of 8 months of age (n: 6 for each sex) were used, in which ANAs and renal functional compromise (proteinuria and increased creatinine) have been described, being the controls (C) mice of both sexes of the B6 strain (n: 6 per sex). The methodologies used for the evaluation of macroautophagy were: transmission electron microscopy (4% Karnovsky fixative, embedding in epoxy resins, contrast with heavy metals, observation in a Zeiss Leo 906-E electron microscope) and immunohistochemistry in paraffin for Beclina-1 and LC3, determining the number of immunopositive cells per tubule, counting between 710 and 790 tubules per control and experimental group. Statistics: mean and SD, Mann-Whitney test (two groups quantitative variable, unpaired, without normal distribution). p<0.05. Protocol approved by CICUAL-University of San Martín. Histopathological evaluation demonstrated, in both male and female NL-like mice, immune complex-mediated mesangial proliferative glomerulonephritis and chronic tubulointerstitial nephritis. The ultrastructural analysis of the experimental groups, unlike the control group and regardless of sex, demonstrated vesicular elements in tubules compatible with macroautophagic vesicles. Both Beclin-1 and LC3 were immunodetected in the renal tubular epithelium, with a significantly higher number of immunopositive cells in the NL-like experimental model compared to the control (example Beclin 1 male C:5±3; NL-like:20±7 *, female C:7±2, similar-NL:25±4** *p<0.05 vs C and **p<0.05 vs C). We can conclude that in the NL-like model the macroautophagy mechanism is increased compared to control animals, possibly linked to the proteinuric state of this strain, and may also contribute to the chronic interstitial damage described. El lupus eritematoso sistémico (LES) es una enfermedad autoinmunitaria compleja y frecuente, presentándose con nefritis lúpica (NL) en 30-60% de los pacientes. La macroautofagia, mecanismo de degradación de contenidos intracelulares, se encuentra desregulada en numerosas enfermedades metabólicas, neurodegenerativas, cáncer y autoinmunes. Si bien se ha demostrado que la autofagia está involucrada en su patogénesis, en qué medida se encuentra presente en los órganos blanco sigue siendo incierto. Nos propusimos analizar y describir la presencia de macroautofagia en un modelo experimental símil-NL. Se utilizaron ratones macho y hembra knockout para Galectina-3 de 8 meses de edad (n:6 por cada sexo), en quienes se han descripto ANAs y compromiso funcional renal (proteinuria e incremento de creatinina), siendo los controles (C) ratones de ambos sexos de la cepa B6 (n:6 por sexo). Las metodologías utilizadas para la evaluación de la macroautofagia fueron: microscopía electrónica de transmisión (fijador Karnovsky 4%, inclusión en resinas epoxi, contraste con metales pesados, observación en un microscopio electrónico Zeiss Leo 906-E) e inmunohistoquímica en parafina para Beclina-1 y LC3, determinándose el número de células inmunopositivas por túbulo, contándose entre 710 y 790 túbulos por grupo control y experimental. Estadística: media y DS, prueba de Mann-Whitney (dos grupos variable cuantitativa, no pareado, sin distribución normal). p<0.05. Protocolo aprobado por CICUAL-Universidad de San Martín. La evaluación histopatológica demostró, tanto en ratones machos como hembras símil-NL, glomerulonefritis proliferativa mesangial mediada por complejos inmunes y nefritis tubulointersticial crónica. El análisis ultraestructural de los grupos experimentales, a diferencia del grupo control e independientemente del sexo, demostró elementos vesiculares en túbulos compatibles con vesículas macroautofágicas. Tanto Beclina-1 como LC3 se inmunodetectaron en el epitelio tubular renal, con un número de células inmunopositivas significativamente mayor en el modelo experimental símil-NL respecto del control (ejemplo Beclina 1 macho C:5±3; simil-NL:20±7*; hembra C:7±2; simil-NL:25±4**. *p<0,05 vs C y **p<0,05 vs C). Podemos concluir que en modelo símil-NL el mecanismo de macroautofagia se encuentra incrementado respecto a los animales controles, posiblemente vinculado al estado proteinúrico propio de esta cepa, pudiendo además contribuir al daño intersticial crónico descripto. Universidad Nacional Córdoba. Facultad de Ciencias Médicas. Secretaria de Ciencia y Tecnología 2023-10-19 info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion application/pdf https://revistas.unc.edu.ar/index.php/med/article/view/42771 Revista de la Facultad de Ciencias Médicas de Córdoba.; Vol. 80 (2023): Suplemento JIC XXIV Revista de la Facultad de Ciencias Médicas de Córdoba; Vol. 80 (2023): Suplemento JIC XXIV Revista da Faculdade de Ciências Médicas de Córdoba; v. 80 (2023): Suplemento JIC XXIV 1853-0605 0014-6722 spa https://revistas.unc.edu.ar/index.php/med/article/view/42771/42807 Derechos de autor 2023 Universidad Nacional de Córdoba http://creativecommons.org/licenses/by-nc/4.0 |