RESUMEN
A kind of commercial hybrid carrageenan (HC)/agarose composite sponge containing κ-, µ-, ι-, and ν-carrageenan, which could turn into hydrogel and release carrageenan at human body temperature was fabricated for immune stimulation and modulation. Release behavior demonstrated that the hybrid carrageenan contained sponge was mechanically stable and could release carrageenan constantly. RT-PCR and ELISA experiments showed that the leaching liquor of the sponge could stimulate RAW264.7 from M0 state to a polarized state by secreting more anti-inflammatory factor IL-10 than pro-inflammatory ones, such as, IL-6 and TNF-α. Transwell experiments also indicated that the leaching liquor could promote the proliferation of NIH-3T3 by stimulating RAW264.7 of M0 state after 7 days. Results of particle size and intracellular concentration analyses suggested that the released carrageenan might enter into the cellular interior of RAW264.7 in the form of microgels or protein complexes. The sponge would be a promising candidate for skin wound dressing.
Asunto(s)
Carragenina/química , Factores Inmunológicos/farmacología , Sefarosa/química , Animales , Vendajes , Proliferación Celular/efectos de los fármacos , Hidrogeles , Interleucina-10/metabolismo , Interleucina-6 , Fenómenos Mecánicos , Ratones , Tamaño de la Partícula , Células RAW 264.7 , Factor de Necrosis Tumoral alfa , Cicatrización de Heridas/efectos de los fármacosRESUMEN
Polycomb group (PcG) proteins are crucial chromatin regulators during development. H2AK119ub1 (H2Aub) and H3K27me3 are catalyzed by Polycomb-repressive complex 1 and 2 (PRC1/2) respectively, and they largely overlap in the genome due to mutual recruitment of the two complexes. However, it is unclear whether PRC1/H2Aub and PRC2/H3K27me3 can also function independently. By developing an ultra-sensitive carrier-DNA-assisted chromatin immunoprecipitation sequencing method termed CATCH-Seq, we generated allelic H2Aub profiles in mouse gametes and early embryos. Our results revealed an unexpected genomewide decoupling of H2Aub and H3K27me3 in mouse preimplantation embryos, where H2Aub but not H3K27me3 was enriched at PcG targets while only H3K27me3 was deposited in the broad distal domains associated with DNA methylation-independent non-canonical imprinting. These observations suggest that H2Aub represses future bivalent genes during early embryogenesis without H3K27me3, but it is not required for the maintenance of non-canonical imprinting, which is mediated by maternal H3K27me3. Thus, our study reveals the distinct depositions and independent functions of H2Aub and H3K27me3 during early mammalian development.