RESUMO
Efforts are made to enhance the inherent potential of extracellular vesicles (EVs) by utilizing 3D culture platforms and engineered strategies for functional cargo-loading. Three distinct types of adipose mesenchymal stem cells-derived EVs (ADSCs-EVs) are successfully isolated utilizing 3D culture platforms consisting of porous gelatin methacryloyl (PG), PG combined with sericin methacryloyl (PG/SerMA), or PG combined with chondroitin sulfate methacryloyl (PG/ChSMA). These correspond to PG-EVs, PG/SerMA-EVs, and PG/ChSMA-EVs, respectively. Unique microRNA (miRNA) profiles are observed in each type of ADSCs-EVs. Notably, PG-EVs encapsulate higher levels of hsa-miR-455-3p and deliver more hsa-miR-455-3p to chondrocytes, which results in the activation of the hsa-miR-455-3p/PAK2/Smad2/3 axis and the subsequent hyaline cartilage regeneration. Furthermore, the functionality of PG-EVs is optimized through engineered strategies, including agomir/lentivirus transfection, electroporation, and Exo-Fect transfection. These strategies, referred to as Agomir-EVs, Lentivirus-EVs, Electroporation-EVs, and Exo-Fect-EVs, respectively, are ranked based on their efficacy in encapsulating hsa-miR-455-3p, delivering hsa-miR-455-3p to chondrocytes, and promoting cartilage formation via the hsa-miR-455-3p/PAK2/Smad2/3 axis. Notably, Exo-Fect-EVs exhibit the highest efficiency. Collectively, the 3D culture conditions and engineered strategies have an impact on the miRNA profiles and cartilage regeneration capabilities of ADSCs-EVs. The findings provide valuable insights into the mechanisms underlying the promotion of cartilage regeneration by ADSCs-EVs.
Assuntos
Vesículas Extracelulares , Cartilagem Hialina , Células-Tronco Mesenquimais , MicroRNAs , Engenharia Tecidual , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/metabolismo , MicroRNAs/genética , MicroRNAs/metabolismo , Vesículas Extracelulares/metabolismo , Vesículas Extracelulares/química , Humanos , Engenharia Tecidual/métodos , Cartilagem Hialina/metabolismo , Regeneração , Tecido Adiposo/citologia , Animais , Técnicas de Cultura de Células em Três Dimensões/métodos , Condrócitos/citologia , Condrócitos/metabolismo , Proteína Smad2/metabolismo , Células Cultivadas , Gelatina/química , Proteína Smad3/metabolismo , Camundongos , CondrogêneseRESUMO
To discover potent insecticides targeting ryanodine receptors (RyRs), a series of novel anthranilic diamides analogues (12a-12u) containing N-substituted phenylpyrazole were designed and synthesized. These compounds were characterized by (1)H NMR, (13)C NMR, and HRMS, and the structure of compound 12u was confirmed by X-ray diffraction. Their insecticidal activities indicated that these compounds displayed moderate to excellent activities. In particular, 12i showed 100 and 37% larvicidal activities against oriental armyworm (Mythimna separata) at 0.25 and 0.05 mg L(-1), equivalent to that of chlorantraniliprole (100%, 0.25 mg L(-1); and 33%, 0.05 mg L(-1)). The activity of 12i against diamondback moth (Plutella xylostella) was 95% at 0.05 mg L(-1), whereas the control was 100% at 0.05 mg L(-1). The calcium-imaging technique experiment results showed that the effects of 12i on the intracellular calcium ion concentration ([Ca(2+)]i) in neurons were concentration-dependent. After the central neurons of Helicoverpa armigera were dyed by loading with fluo-5N and treated with 12i, the free calcium released in endoplasmic reticulum indicated the target of compound 12i is RyRs or IP3Rs. The activation of RyRs by natural ryanodine completely blocked the calcium release induced by 12i, which indicated that RyRs in the central neurons of H. armigera third-instar larvae is the possible target of compound 12i.
Assuntos
Agonistas dos Canais de Cálcio/síntese química , Inseticidas/química , Isoxazóis/química , Canal de Liberação de Cálcio do Receptor de Rianodina/química , Animais , Agonistas dos Canais de Cálcio/química , Agonistas dos Canais de Cálcio/metabolismo , Diamida , Desenho de Fármacos , Proteínas de Insetos/antagonistas & inibidores , Proteínas de Insetos/química , Proteínas de Insetos/metabolismo , Inseticidas/síntese química , Inseticidas/farmacologia , Larva/efeitos dos fármacos , Larva/crescimento & desenvolvimento , Estrutura Molecular , Mariposas/efeitos dos fármacos , Mariposas/crescimento & desenvolvimento , Canal de Liberação de Cálcio do Receptor de Rianodina/metabolismo , Relação Estrutura-Atividade , Difração de Raios XRESUMO
In this study, the chemical constituentsfrom Valeriana amurensis AD-effective fraction were investigated based on the effect of Valeriana amurensis on Alzheimer's disease (AD) in previous study. Valeriana amurensis was extracted with 75% ethanol and the obtained extract were extracted and subjected to AB-8 macroporous resin column to obtain the AD-effective fraction of Valeriana amurensis. 9 compounds (1-9) were isolated with silica gel, ODS column chromatography and preparative HPLC. The structures of these compounds were determined as 6-hydroxy-7-(hydroxymethyl)-4-methylenehexahydrocyclopenta[c]-pyran-1(3H)-one (1), suspensolide F (2), loganin(3), α-morroniside(4), ß-morronisid (5), partinovalerosidate (6), zansiumloside A (7), (-)-angelicoidenol-2-O-ß-D-glucopyranoside (8), citroside A (9). Compounds 6-9 were isolated from the valerian genus for the first time and further investigated the anti-AD effect of compounds 1-9 in vitro found that compound 2 and 6 protected the PC12 cells from injury significantly.
