RESUMO
Periodontal ligament stem cells (PDLSCs) show plasticity towards the adipogenic lineage; however, little has been done on the participation of epigenetic mechanisms. Histone acetylation is a dynamic process, though balanced by histone acetyltransferases (HATs) and histone deacetylases (HDACs) activities. This process can be halted by HDACs inhibitors, such as trichostatin A (TSA) and valproic acid (VPA). This study aimed to determine the role of HDACs class I in adipogenic differentiation of PDL cells. PDLSCs were treated with TSA at concentrations of 100, 200, and 250 nM, or VPA at 1, 4 and 8 mM. Cell viability was assessed using MTT assays. Gene expression of pluripotency markers (NANOG, OCT4, SOX2), HAT genes (p300, GCN5), and HDACs genes (HDAC1-3) was analyzed by RT-qPCR. Adipogenic differentiation was evaluated via oil red O staining, and acetylation of histone H3 lysine 9 (H3K9ac) was examined by Western blot. VPA treatment resulted in a 60% reduction in cell proliferation, compared to a 50% when using TSA. Cell viability was not affected by either inhibitor. Furthermore, both TSA and VPA induced adipogenic differentiation, through an increase in the deposition of lipid droplets and in GCN5 and p300 expression were observed. Western blot analysis showed that TSA increased H3K9ac levels on adipogenic differentiation of PDLSCs. These findings highlight the potential of HDAC inhibitors as a tool for modulating H3K9 acetylation status and thus influencing adipogenic differentiation of PDLCs.
Assuntos
Adipogenia , Diferenciação Celular , Sobrevivência Celular , Inibidores de Histona Desacetilases , Ligamento Periodontal , Ácido Valproico , Humanos , Ligamento Periodontal/citologia , Ligamento Periodontal/efeitos dos fármacos , Inibidores de Histona Desacetilases/farmacologia , Adipogenia/efeitos dos fármacos , Adipogenia/genética , Ácido Valproico/farmacologia , Diferenciação Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Acetilação/efeitos dos fármacos , Histona Desacetilases/metabolismo , Histona Desacetilases/genética , Ácidos Hidroxâmicos/farmacologia , Células Cultivadas , Histonas/metabolismo , Proliferação de Células/efeitos dos fármacos , Células-Tronco/efeitos dos fármacos , Células-Tronco/citologia , Células-Tronco/metabolismoRESUMO
El objetivo de este trabajo fue la selección de cepas nativas del género Bacillus con actividad quitinolíticay proteolítica, en suelo tropical en la costa de Oaxaca, México. Se aislaron 150 cepas, de las cuales 22fueron seleccionadas por presentar actividad quitinolítica y proteolítica. Dicha actividad se evaluó porla formación de halo de hidrólisis alrededor de la colonia en medios de cultivo suplementados con quitinacoloidal al 5% y leche descremada al 1% respectivamente. Las cepas LUM B001, B003, B013, B015y B065 presentaron mayor actividad quitinolítica y proteolítica, por lo que tienen el potencial para serevaluadas en control biológico de hongos fitopatógenos. Se encontró al género Bacillus distribuido ensuelos cultivados y no cultivados, no se encontraron diferencias estadísticas según el cultivo establecido(P<0,05), sin embargo se encontraron diferencias significativas (P<0,05) entre las zonas estudiadas, correspondiendolas menores recuperaciones de cepas a los terrenos del municipio de Tututepec, Oaxaca.
This work was aimed at selecting native strains from the Bacillus genus having chitinolytic and proteolytic activityfrom soil from the tropical coast of Oaxaca, Mexico. 150 strains were isolated, 22 of which were selectedas they presented chitinolytic and proteolytic activity. Such activity was assessed by the formation of a hydrolysishalo around the colony in culture media supplemented with 5% colloidal chitin and 1% skimmed milk.The LUM B001, B003, B013, B015 and B065-chitin strains presented higher quitinolytic and proteolytic activity,thereby having the potential for being evaluated in the biological control of phytopathogenic fungi. TheBacillus genus was found in cultivated and uncultivated soils; no statistical differences were found accordingto established crop (p <0.05); however, significant differences (p <0.05) were found between the areas beingstudied regarding the smaller amount of strains collected from land in the municipality of Tututepec, Oaxaca.
