RESUMEN
The use of DNA aggregates could be a promising strategy for the molecular imaging of biological functions. Herein, phosphorescent oligodeoxynucleotides were designed with the aim of visualizing oxygen fluctuation in tumor cells. DNA-ruthenium conjugates (DRCs) that consisted of oligodeoxynucleotides, a phosphorescent ruthenium complex, a pyrene unit for high oxygen responsiveness, and a nitroimidazole unit as a tumor-targeting unit were prepared. In general, oligonucleotides have low cell permeability because of their own negative charges; however, the DRC formed aggregates in aqueous solution due to the hydrophobic pyrene and nitroimidazole groups, and smoothly penetrated the cellular membrane to accumulate in tumor cells in a hypoxia-selective manner. The oxygen-dependent phosphorescence of DRC in cells was also observed. In vivo experiments revealed that aggregates of DRC accumulated in hypoxic tumor tissue that was transplanted into the left leg of mice, and showed that oxygen fluctuations in tumor tissue could be monitored by tracking of the phosphorescence emission of DRC.
Asunto(s)
Sustancias Luminiscentes/química , Oligodesoxirribonucleótidos/química , Oxígeno/análisis , Células A549 , Animales , Complejos de Coordinación/síntesis química , Complejos de Coordinación/química , Complejos de Coordinación/efectos de la radiación , Colorantes Fluorescentes/química , Humanos , Concentración de Iones de Hidrógeno , Luz , Sustancias Luminiscentes/síntesis química , Sustancias Luminiscentes/efectos de la radiación , Ratones Endogámicos BALB C , Imagen Molecular , Nitroimidazoles/síntesis química , Nitroimidazoles/química , Oligodesoxirribonucleótidos/síntesis química , Oligodesoxirribonucleótidos/efectos de la radiación , Oxazinas/química , Oxígeno/química , Pirenos/síntesis química , Pirenos/química , Rutenio , Hipoxia TumoralRESUMEN
The steroid hormones synthesized by the male gonads play diverse roles in biological processes. Androgens, the primary hormones produced by the male gonads, are key regulators of fat homeostasis, hence androgen-deprivation therapies often induce obesity. However, the molecular mechanism by which male gonadal dysfunction leads to obesity remains unclear, because results from animal studies regarding fat accumulation in the context of gonadal defects do not reflect clinical findings. Here, we investigated the mechanism underlying the development of obesity in animals with male gonadal dysfunction by analyzing the long-term physiological changes in adult male mice with surgical castration. Nine weeks after surgery, white adipose tissue (WAT) mass was higher in the castrated (Cas) mice than in sham-operated (Sham) mice. In addition, castration induced hyperlipidemia and hyperglycemia. However, genes involved in lipid metabolism, including hormone-sensitive lipase, were unchanged in the adipose tissue of the Cas mice, despite the increase in WAT. In contrast, a hepatic gluconeogenesis gene, glucose-6-phosphatase, was significantly upregulated in the Cas mice than in Sham mice. Our findings suggest that long-term hypogonadism in mice mimics the effects in humans, and a potential molecular basis for the induction of obesity in this model is impairment of hepatic gluconeogenesis.
Asunto(s)
Gluconeogénesis , Hipogonadismo/complicaciones , Obesidad/etiología , Tejido Adiposo Blanco/patología , Animales , Glucemia/análisis , Peso Corporal , Ingestión de Alimentos , Ácidos Grasos no Esterificados/sangre , Hiperglucemia/sangre , Hiperglucemia/etiología , Hiperlipidemias/sangre , Hiperlipidemias/etiología , Hipogonadismo/sangre , Hígado/metabolismo , Hígado/patología , Masculino , Ratones Endogámicos C57BL , Obesidad/sangre , Orquiectomía , Tamaño de los Órganos , Triglicéridos/sangreRESUMEN
Bofutsushosan (BTS; fangfengtongshengsan in Chinese) is a formula in traditional Japanese Kampo and Chinese medicine comprising 18 crude drugs and used to treat obesity and metabolic syndrome. In our previous study, BTS boiling water extract inhibited the uptake of fructose absorbed via glucose transporter 5 into cultured cells. In this study, the inhibitory effect of BTS extract on the absorption of fructose from the intestine was investigated in vivo. The extract of BTS was orally administered to rats at doses equivalent to 25-fold of the daily dose for humans. One minute after sample administration, fructose was orally administered and blood samples were collected from the jugular vein 0.5, 1, 1.5, 2, and 4 h after the administration of fructose. The absorption of fructose from the intestine was significantly reduced by treatment with BTS extract, and this in vivo study reproduced previous in vitro results. Subsequently, the blood samples were collected from the portal vein 30 min after the oral administration of fructose in mice. BTS extract significantly reduced fructose absorption in mice, and compared the effect of modified BTS samples by removing one to several crude drugs from BTS. We found that the dried rhizome of Rheum palmatum (RR) significantly contributed to the inhibitory effect of BTS on fructose absorption. We found sennoside A to be the active ingredient of RR for the inhibition of fructose absorption, and that its effect almost saturated at a dose of 3 mg/kg. These results support the action mechanisms of BTS when used for the treatment of obesity in clinics and drug stores.
Asunto(s)
Medicamentos Herbarios Chinos , Fructosa , Humanos , Ratones , Ratas , Animales , Medicamentos Herbarios Chinos/farmacología , Medicamentos Herbarios Chinos/uso terapéutico , Obesidad , Senósidos/uso terapéuticoRESUMEN
The incidence of sensitive skin with stinging and itch following chemical exposure in products such as cosmetics is increasing, but molecular mechanisms underlying this pathophysiology remain understudied. Here we performed transcriptional analysis of reconstructed human epidermis (RHE) 1, 6, and 24â¯h following topical lactic acid (LA) application, a known inducer of the sensitive skin reaction. Since little is known about the specific role of keratinocyte transcriptional changes in mediating stinging and itch, we performed pathway analysis using several publically available databases and then focused on significantly changed transcripts involved in stress responses and itch signaling using the Comparative Toxicogenomics Database. LA treatment induced damage-associated genes HSPA1A, DDIT3, IL1A, and HMGB2. Neurotrophic factors including BDNF, ARTN, PGE2, and chemokines were also upregulated. Stimulation of the RHE with 5% LA did not reduce cell viability, but reduced the trans-epidermal electric resistance, suggesting barrier dysfunction. Accordingly, skin barrier formation genes such as filaggrins (FLG, FLG2) and corneodesmosin (CDSN) were downregulated. To our knowledge, this is the first study focusing on transcriptional changes underlying the stinging response of keratinocytes upon LA stimulation. While follow-up research is needed, this study provides new insight into the mechanisms underlying the sensitive skin reaction.