Effect of caspase-dependent pathway on apoptosis of human gingival fibroblasts induced by advanced glycation end products / 中华口腔医学杂志
Chinese Journal of Stomatology
; (12): 673-677, 2011.
Article
en Zh
| WPRIM
| ID: wpr-306364
Biblioteca responsable:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of synthesized advanced glycation end products (AGE) on apoptosis of human gingival fibroblasts (HGF) and the possible role of caspase-dependent pathway in the process of AGE-induced apoptosis.</p><p><b>METHODS</b>HGF were incubated with AGE-human serum albumin (AGE-HSA). The activity of caspase-8, caspase-9 and caspase-3 were detected by microplate reader after 12 and 24 hours. HGF were incubated with caspase inhibitors for 1 hour, and then incubated with AGE-HSA for 24 hours, HGF was first stained by Hoechst33258 and observed under inverted microscope, and then double stained by annexin V and propidine iodide (PI) and observed by flow cytometry (FCM). The activity of caspase-3 was determined by caspase-3 assay kit and observed by microplate reader.</p><p><b>RESULTS</b>Caspases activity of caspase-8, -9, -3 was 0.1097 ± 0.0051, 0.0965 ± 0.0051 and 0.1280 ± 0.0103 after 12 h of incubation with AGE-HSA and HGF, respectively, and 0.1558 ± 0.0053, 0.1308 ± 0.0035 and 0.1954 ± 0.0051 after 24 h of incubation with AGE-HSA and HGF, respectively (P < 0.05). Positive cells number was 247.7 ± 32.4, 200.1 ± 14.6, 154.1 ± 14.4 and 131.3 ± 14.6 in caspase inhibitor groups by Hoechst33258 staining, respectively. Apoptotic rate was (25.57 ± 2.20)%, (38.87 ± 3.31)%, (17.17 ± 2.24)% and (14.73 ± 2.48)% in caspase inhibitor groups by annexin V-PI staining, respectively. The difference between different groups was significant (P < 0.05). Caspase-3 activity was reduced to 0.1274 ± 0.0076, 0.1465 ± 0.0062, 0.1044 ± 0.0051 in caspase inhibitor groups, respectively. The difference between different groups was significant (P < 0.05).</p><p><b>CONCLUSIONS</b>Apoptosis of HGF induced by AGE-HSA may be mainly through activation of caspase-dependant pathway in which cytoplasmic pathway may play a predominant role.</p>
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Índice:
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Asunto principal:
Oligopéptidos
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Farmacología
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Albúmina Sérica
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Transducción de Señal
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Células Cultivadas
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Productos Finales de Glicación Avanzada
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Apoptosis
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Caspasas
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Biología Celular
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Caspasa 3
Límite:
Adolescent
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Adult
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Child
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Humans
Idioma:
Zh
Revista:
Chinese Journal of Stomatology
Año:
2011
Tipo del documento:
Article