RESUMO
Chlorogenic acid (CGA) is an effective phenolic antioxidant that can scavenge hydroxyl radicals and superoxide anions. Herein, the protective effects and mechanisms leading to CGA-induced porcine parthenogenetic activation (PA) in early-stage embryos were investigated. Our results showed that 50 µM CGA treatment during the in vitro culture (IVC) period significantly increased the cleavage and blastocyst formation rates and improved the blastocyst quality of porcine early-stage embryos derived from PAs. Then, genes related to zygotic genome activation (ZGA) were identified and investigated, revealing that CGA can promote ZGA in porcine PA early-stage embryos. Further analysis revealed that CGA treatment during the IVC period decreased the abundance of reactive oxygen species (ROS), increased the abundance of glutathione and enhanced the activity of catalase and superoxide dismutase in porcine PA early-stage embryos. Mitochondrial function analysis revealed that CGA increased mitochondrial membrane potential and ATP levels and upregulated the mitochondrial homeostasis-related gene NRF-1 in porcine PA early-stage embryos. In summary, our results suggest that CGA treatment during the IVC period helps porcine PA early-stage embryos by regulating oxidative stress and improving mitochondrial function.
Assuntos
Ácido Clorogênico , Técnicas de Cultura Embrionária , Desenvolvimento Embrionário , Mitocôndrias , Estresse Oxidativo , Partenogênese , Espécies Reativas de Oxigênio , Animais , Estresse Oxidativo/efeitos dos fármacos , Partenogênese/efeitos dos fármacos , Mitocôndrias/efeitos dos fármacos , Técnicas de Cultura Embrionária/veterinária , Ácido Clorogênico/farmacologia , Desenvolvimento Embrionário/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Blastocisto/efeitos dos fármacos , Suínos , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Antioxidantes/farmacologia , Feminino , Glutationa/metabolismoRESUMO
Lysine-specific demethylase 1 (LSD1) stands as the pioneering histone demethylase uncovered, proficient in demethylating H3K4me1/2 and H3K9me1/2, thereby governing transcription and participating in cell apoptosis, proliferation, or differentiation. Nevertheless, the complete understanding of LSD1 during porcine early embryonic development and the underlying molecular mechanism remains unclear. Thus, we investigated the mechanism by which LSD1 plays a regulatory role in porcine early embryos. This study revealed that LSD1 inhibition resulted in parthenogenetic activation (PA) and in vitro fertilization (IVF) embryo arrested the development, and decreased blastocyst quality. Meanwhile, H3K4me1/2 and H3K9me1/2 methylase activity was increased at the 4-cell embryo stage. RNA-seq results revealed that autophagy related biological processes were highly enriched through GO and KEGG pathway analyses when LSD1 inhibition. Further studies showed that LSD1 depletion in porcine early embryos resulted in low mTOR and p-mTOR levels and high autophagy and apoptosis levels. The LSD1 deletion-induced increases in autophagy and apoptosis could be reversed by addition of mTOR activators. We further demonstrated that LSD1 inhibition induced mitochondrial dysfunction and mitophagy. In summary, our research results indicate that LSD1 may regulate autophagy and apoptosis through the mTOR pathway and affect early embryonic development of pigs.
Assuntos
Apoptose , Autofagia , Desenvolvimento Embrionário , Histona Desmetilases , Transdução de Sinais , Serina-Treonina Quinases TOR , Animais , Histona Desmetilases/metabolismo , Histona Desmetilases/genética , Suínos/embriologia , Serina-Treonina Quinases TOR/metabolismo , Serina-Treonina Quinases TOR/genética , Desenvolvimento Embrionário/fisiologia , Autofagia/fisiologia , Regulação da Expressão Gênica no Desenvolvimento , Fertilização in vitro/veterináriaRESUMO
Recent studies have shown that sulforaphane (SFN) selectively inhibits the growth of ALDH⺠breast cancer stem-like cells.Herein, a series of SFN analogues were synthesized and evaluated against breast cancer cell lines MCF-7 and SUM-159, and the leukemia stem cell-like cell line KG-1a. These SFN analogues were characterized by the replacement of the methyl group with heterocyclic moieties, and the replacement of the sulfoxide group with sulfide or sulfone. A growth inhibitory assay indicated that the tetrazole analogs 3d, 8d and 9d were significantly more potent than SFN against the three cancer cell lines. Compound 14c, the water soluble derivative of tetrazole sulfide 3d, demonstrated higher potency against KG-1a cell line than 3d. SFN, 3d and 14c significantly induced the activation of caspase-3, and reduced the ALDH⺠subpopulation in the SUM159 cell line, while the marketed drug doxrubicin(DOX) increased the ALDH⺠subpopulation.