RESUMEN
This article describes the physicochemical properties of chitosan-coated liposomes containing skin-protecting agents, coenzyme Q10 and alpha-lipoic acid (CCAL). CCAL had a spherical shell-core structure and liposomes inverted the surface charge from negative to positive after coating with chitosan. Compared with the uncoated liposome, CCAL had higher zeta potential, larger droplet size and long-term stability. Fourier transform infrared spectroscopy (FTIR) study showed that the driving force for chitosan coating the liposomes was enhanced via hydrogen bonding and ionic bond force between the chitosan and the alpha-lipoic acid. While the encapsulation efficiency (EE) of alpha-lipoic acid also increased by interacting with the chitosan shell. In vitro antioxidant activity study showed an excellent hydroxyl radical scavenging activity of CCAL. In vitro release study displayed a sustained drug release, and in vitro penetration studies promoted the accumulation of drugs in rabbit skin.
Asunto(s)
Quitosano , Materiales Biocompatibles Revestidos , Depuradores de Radicales Libres , Absorción Cutánea , Ácido Tióctico , Ubiquinona/análogos & derivados , Administración Cutánea , Animales , Quitosano/química , Quitosano/farmacocinética , Quitosano/farmacología , Materiales Biocompatibles Revestidos/química , Materiales Biocompatibles Revestidos/farmacocinética , Materiales Biocompatibles Revestidos/farmacología , Depuradores de Radicales Libres/química , Depuradores de Radicales Libres/farmacocinética , Depuradores de Radicales Libres/farmacología , Liposomas , Conejos , Ácido Tióctico/química , Ácido Tióctico/farmacocinética , Ácido Tióctico/farmacología , Ubiquinona/química , Ubiquinona/farmacocinética , Ubiquinona/farmacologíaRESUMEN
Cytochrome P450s (CYPs) are widespread proteins that interact with exogenous chemicals from the diet or the environment. CYP9A subfamily genes are important in the silkworm Bombyx mori. We previously reported transcriptional levels of two CYP9A genes in different tissues and their responses to sodium fluoride (NaF). In this study, promoter truncation analysis using a dual-luciferase reporter assay in B. mori ovary cells (BmN) showed that the regions -1,496 to -1,102 bp for CYP9A19, and -1,630 to -1,210 bp for CYP9A22 were essential for basal transcriptional activity. Sequence analysis of these regions revealed several transcriptional regulatory elements but no typical promoter elements. Promoter activities were regulated after NaF induction and with an obvious dose effect. Although the dual-luciferase assay has been widely used to determine the activity of a given promoter in cell lines, problems with it still exist. Our results indicate that both plasmid size and construct protocols affect the experimental results.
Asunto(s)
Bombyx/enzimología , Sistema Enzimático del Citocromo P-450/genética , Regulación Enzimológica de la Expresión Génica , Proteínas de Insectos/genética , Animales , Secuencia de Bases , Bombyx/genética , Línea Celular , Clonación Molecular , Técnicas de Silenciamiento del Gen , Genes Reporteros , Luciferasas de Luciérnaga/biosíntesis , Luciferasas de Luciérnaga/genética , Luciferasas de Renilla/biosíntesis , Luciferasas de Renilla/genética , Datos de Secuencia Molecular , ARN Interferente Pequeño/genética , Análisis de Secuencia de ADN , Fluoruro de Sodio/farmacología , TransfecciónRESUMEN
Glutathione S-transferases (GSTs) are a multifunctional super gene family, some of which play an important role in insecticide resistance. In this research, we used a real-time quantitative RT-PCR method, and a novel strategy, to measure the transcriptional level per gene copy using an exogenous RNA reference and DNA reference. The transcription levels of six BmGST genes in different tissues of fifth instar Bombyx mori larvae and their responses to insecticide and fluoride were investigated. The results show different levels and patterns of expression of the different BmGSTs in the various tissues observed. The BmGSTs can be induced by insecticide and fluoride, but their responses to each are different. The results of this research are helpful in studying the tissue-specific expression of BmGSTs in Bombyx mori, and in developing new pesticide resistant silkworm varieties.