Activation of Satellite Cells in the Intercostal Muscles of Patients With Chronic Obstructive Pulmonary Disease

Martínez-Llorens, Juana; Casadevall, Carme; Lloreta, Josep; Orozco-Levi, Mauricio; Barreiro, Esther; Broquetas, Joan; Gea, Joaquim

doi:10.1016/S1579-2129(08)60038-5

Article information

Objective

The respiratory muscles of patients with chronic obstructive pulmonary disease (COPD) display evidence of structural damage in parallel with signs of adaptation. We hypothesized that this can only be explained by the simultaneous activation of satellite cells. The aim of this study was to analyze the number and activation of those cells along with the expression of markers of microstructural damage that are frequently associated with regeneration.

Patients and methods

The study included 8 patients with severe COPD (mean [SD] forced expiratory volume in 1 second, 33% [9%] of predicted) and 7 control subjects in whom biopsies were performed of the external intercostal muscle. The samples were analyzed by light microscopy to assess muscle fiber phenotype, electron microscopy to identify satellite cells, and real-time polymerase chain reaction to analyze the expression of the following markers: insulin-like growth factor 1, mechano growth factor, and embryonic and perinatal myosin heavy chains (MHC) as markers of microstructural damage; Pax-7 and m-cadherin as markers of the presence and activation of satellite cells, respectively; and MHC-I, IIa, and IIx as determinants of muscle fiber phenotype.

Results

The patients had larger fibers than healthy subjects (54 [6] vs 42 [4] μm2; P < .01) with a similar or slightly increased proportion of satellite cells, as measured by ultrastructural analysis (4.3% [1%] vs 3.7% [3.5%]; P > .05) or expression of Pax-7 (5.5 [4.1] vs 1.6 [0.8] arbitrary units [AU]; P < .05). In addition, there was greater activation of satellite cells in the patients, as indicated by increased expression of m-cadherin (3.8 [2.1] vs 1.0 [1.2] AU; P=.05). This was associated with increased expression of markers of microstructural damage: insulin-like growth factor 1, 0.35 (0.34) vs 0.09 (0.08) AU (P < .05); mechano growth factor, 0.45 (0.55) vs 0.13 (0.17) AU (P=.05). CONCLUSIONS: The intercostal muscles of patients with severe COPD show indirect signs of microstructural damage accompanied by satellite cell activation. This suggests the presence of ongoing cycles of lesion and repair that could partially explain the maintenance of the structural properties of the muscle.

Key words:

COPD

Muscle dysfunction

Damage

Repair

Satellite cells

Objetivo

Los músculos respiratorios de los pacientes con enfermedad pulmonar obstructiva crónica (EPOC) presentan lesiones estructurales, que coexisten con signos de adaptación. Nuestra hipótesis es que esto sólo puede explicarse si se produce simultáneamente la activación de sus células satélite. El propósito del presente trabajo ha sido valorar el número y la eventual activación de dichas células, así como la expresión de marcadores de microlesión estructural, ligados a la regeneración.

Pacientes y métodos

Se incluyó en el estudio a 8 pacientes con EPOC grave -media ± desviación estándar del volumen espiratorio forzado en el primer segundo: un 33 ± 9% del valor de referencia-y a 7 controles, a quienes se realizó una biopsia del músculo intercostal externo. La muestra se analizó mediante microscopia óptica (fenotipo fibrilar), electrónica (células satélite) y técnica de reacción en cadena de la polimerasa en tiempo real (marcadores de microlesión: factor de crecimiento similar a la insulina de tipo 1, factor de crecimiento mecánico e isoformas de cadenas pesadas de miosina [MyHC] embrionaria e isoformas de perinatal; de presencia y activación de células satélite: Pax-7 y m-caderina, respectivamente; y condicionantes del fenotipo fibrilar: MyHC-I, IIa y IIx).

Resultados

Los pacientes tuvieron unas fibras mayores que los sujetos sanos (54 ± 6 frente a 42 ± 4 |a, m2; p < 0,01), con una población de células satélite conservada o ligeramente incrementada (cuantificación ultraestructural: 4,3 ± 1% frente al 3,7 ± 3,5%, p no significativa; Pax-7: 5,5 ± 4,1 frente a 1,6 ± 0,8, unidades arbitrarias [ua], respectivamente, p < 0,05) y una mayor activación (m-caderina: 3,8 ± 2,1 frente a 1,0 ± 1,2 ua; p = 0,05). Esto se asociaba a valores aumentados de marcadores de microlesión (factor de crecimiento similar a la insulina de tipo 1: 0,35 ± 0,34 frente a 0,09 ± 0,08 ua, p < 0,05; factor de crecimiento mecánico: 0,45 ± 0,55 frente a 0,13 ± 0,17 ua, p = 0,05).

Conclusiones

Los músculos intercostales de pacientes con EPOC grave muestran signos indirectos de microlesión, acompañados de la activación de sus células satélite. Esto apunta a la presencia de ciclos continuados de lesión y reparación, lo que podría explicar parcialmente la conservación de sus propiedades estructurales.

Palabras clave:

EPOC

Disfunción muscular

Daño

Reparación

Células satélite

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This study was supported by funds from the European Union (Fifth Framework Programme, reference QLRT-2000-00417), the Spanish Ministry of Science and Technology (National R+D Plan, reference SAF 2001-0426), the Spanish Ministry of Health (Red RESPIRA, reference RTIC C03/11, Health Research Fund, Carlos III Health Institute), the Spanish Society of Pulmonology and Thoracic Surgery (SEPAR), and the Generalitat de Cataluña (2005SGR01060).

J. Martínez-Llorens was supported by institutional funding from SEPAR, the Catalan Society of Pulmonology (SOCAP), and the Municipal Institute for Medical Research (IMIM, grants for residents, 2002 and 2003).

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