RESUMEN
GNE myopathy, caused by biallelic mutations in the GNE gene, is characterized by initial ankle dorsiflexor weakness and rimmed vacuoles in the muscle histopathology, resulting in reduced sialic acid production. Sialyllactose is a source of sialic acid. We performed a pilot clinical trial to analyze the pharmacokinetic properties of 6'-sialyllactose (6SL) and evaluated the safety, and efficacy of oral 6SL in patients with GNE myopathy. Ten participants were in the pharmacokinetic study, and 20 in the subsequent clinical trial. For the pharmacokinetic study, participants were administered either 3 g (low-dose) or 6 g (high-dose) of 6SL in a single dose. Plasma concentrations of 6SL, sialic acid, and sialic acid levels on the surface of red blood cells were periodically assessed in blood samples. Patients were randomly allocated to test (low- and high-dose groups) or placebo groups for the trial. Motor function, ambulation, plasma 6SL and sialic acid concentrations, GNE myopathy-functional activity scale scores, and MRI findings were assessed. 6SL was well tolerated, except for self-limited gastrointestinal discomfort. Free sialic acid in both low- and high-dose groups significantly increased at 6 and 12 weeks, but not in the placebo group. In the high-dose group, proximal limb powers improved with daily 6SL. Considering the fat fraction on muscle MRI, results in the high-dose group were superior to those in the low-dose group. 6SL may be a good candidate for GNE myopathy therapeutics as it induces an increase or reduces the decrease in limb muscle power, attenuates muscle degeneration, and improves the biochemical properties of sialic acid.
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Miopatías Distales , Ácido N-Acetilneuramínico , Humanos , Ácido N-Acetilneuramínico/uso terapéutico , Proyectos Piloto , Miopatías Distales/tratamiento farmacológico , Miopatías Distales/genética , Miopatías Distales/patología , Resultado del Tratamiento , Músculo Esquelético/patología , MutaciónRESUMEN
BACKGROUND: Gamisoyosan (GSS) is an herbal formula which has been used to treat women's diseases for several hundred years in Korea. GSS is one of the three most common prescriptions among women and is used to treat menopausal symptoms. Fatty liver disease is also common in postmenopausal women and can precede more severe diseases, such as steatohepatitis. The present study compared the effects of GSS on fatty liver using three different formulae, Dongui-Bogam (KIOM A), Korean Pharmacopeia (KIOM B) and Korean National Health Insurance (KIOM C). METHODS: In oleic acid-induced HepG2 fatty liver cells, cellular lipid accumulation, triglycerides and total cholesterol were measured after treatment with three GSS formulae and simvastatin as a positive control. To investigate the phytoestrogen activity of GSS, MCF-7 cells were treated with GSS, and hormone levels were quantified. Also, qualitative analysis was performed with UPLC. RESULTS: All types of GSS decreased cellular lipid accumulation. KIOM A was slightly less effective than the other two GSS formulae. KIOM B and KIOM C decreased cellular triglycerides more effective than simvastatin, but KIOM A did not affect cellular triglycerides. Cellular total cholesterol was decreased by all GSS and simvastatin. GSS showed phytoestrogen activity in MCF-7 cells. From the UPLC analysis data, geniposide, paeoniflorin and glycyrrhizin were detected form three GSS formulae. CONCLUSION: These results suggest that all GSS formulae have a beneficial effect on fatty liver disease during menopause and that differences of formula have no effect on the efficacy of the prescription.
RESUMEN
Curcumae Radix (CR) and Glycyrrhizae Radix et Rhizoma (GR) extracts have been used as health supplements in traditional medicine. This study was performed to evaluate the effects of combined CR and GR extracts (CR+GR) on metabolic complications related menopausal symptoms. We found a significant results that CR+GR extracted using ethanol stimulated the growth of MCF-7 cells in estrogen activity and was attenuated in lipid deposition of HepG2 cells treated with MßCD compared to CR and GR treatments each. To investigate the situation, an experimental menopause rat model with dyslipidemia was induced by surgical bilateral ovariectomy (OVX) and high fat high cholesterol (HFHC) diet in female rats. OVX rats fed HFHC (OVX-HFHC) showed a shift in weight gain, elevated serum cholesterol, altered liver enzymatic parameters and enhanced liver injury compared to the NC and HFHC groups. However, administration of CR+GR, in particular 200 or 450 mg/kg/day, inhibited the increase in body weight gain and lipid metabolic disturbances, lowering total cholesterol (TC), triglycerides (TG) and low density lipoprotein cholesterol (LDL-C) compared to the OVX-HFHC group. Furthermore, CR+GR (200 or 450 mg/kg/day) ameliorated the serum levels of the liver enzymes aspartate aminotransferase (AST) and alanine transaminase (ALT) compared to the OVX-HFHC group. Moreover, CR+GR (200 or 450 mg/kg/day) attenuated not only hepatic steatosis but also larger adipocytes. Our study demonstrated that combined treatment with CR and GR attenuated metabolic complications induced by OVX and HFHC diet, suggesting that this effect may regulate and prevent the acceleration of cardiovascular disease (CVD) after menopause.
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The present study was conducted to investigate the effect of Sagunja-tang on the lipid related disease in a rat model of menopausal hyperlipidemia and lipid accumulation in methyl-ß-cyclodextrin-induced HepG2 cells. In in vivo study using menopausal hyperlipidemia rats, Sagunja-tang reduced retroperitoneal and perirenal fat, serum lipids, atherogenic index, cardiac risk factor, media thickness, and nonalcoholic steatohepatitis score, when compared to menopausal hyperlipidemia control rats. In HepG2 cells, Sagunja-tang significantly decreased the lipid accumulation, total cholesterol levels, and low-density/very-low-density lipoprotein levels. Moreover, Sagunja-tang reversed the methyl-ß-cyclodextrin-induced decrease in the protein levels of critical molecule involved in cholesterol synthesis, sterol regulatory element binding protein-2, and low-density lipoprotein receptor and inhibited protein levels of 3-hydroxy-3-methylglutaryl coenzyme A reductase as well as activity. Phosphorylation level of AMP-activated protein kinase was stimulated by Sagunja-tang. These results suggest that Sagunja-tang has effect on inhibiting hepatic lipid accumulation through regulation of cholesterol synthesis and AMPK activity in vitro. These observations support the idea that Sagunja-tang is bioavailable both in vivo and in vitro and could be developed as a preventive and therapeutic agent of hyperlipidemia in postmenopausal females.
RESUMEN
OBJECTIVE: We examined the phytoestrogenic effects of palmiwon on breast carcinoma, lipid accumulation in methyl-ß-cyclodextrin-induced HepG2 cells, and lipid-related diseases in a rat model of menopausal hyperlipidemia. METHODS: E-Screen assay was used to screen for phytoestrogens, especially those with antiestrogenic activity, in MCF-7 cells. Oil Red O staining and intracellular cholesterol analyses were used to quantify cellular cholesterol levels. 3-Hydroxy-3-methyl glutaryl coenzyme A reductase assay was used to measure enzyme activity. The levels of phosphorylated adenosine monophosphate-activated protein kinases and products of genes involved in cholesterol synthesis were measured by Western blot analysis. Thirty rats were either ovariectomized or sham-operated and randomly assigned to four groups (n = 5)-Sham, OVX, OVX-SV, or OVX-PMW (50, 150, or 450 mg/kg) group-for 8 weeks. A number of targets associated with lipid-related diseases were examined to confirm the estrogenic effects of palmiwon. RESULTS: Palmiwon showed antiestrogenic activity in MCF-7 cells. Palmiwon decreased lipid accumulation, total cholesterol levels, and low-density lipoprotein/very-low-density lipoprotein levels in HepG2 cells. Moreover, palmiwon reversed the effects of methyl-ß-cyclodextrin on cholesterol synthesis regulators and inhibited the activity of 3-hydroxy-3-methyl glutaryl coenzyme A reductase. Phosphorylation of adenosine monophosphate-activated protein kinase was stimulated by palmiwon. In ovariectomized rats, palmiwon reduced retroperitoneal and perirenal fat accumulation, serum lipids, atherogenic index, cardiac risk factor score, intima-media thickness, and nonalcoholic steatohepatitis scores. CONCLUSIONS: These results indicate that palmiwon inhibits lipid accumulation without estrogenic activity in the breast. Therefore, palmiwon may have potential as a therapeutic agent for the treatment of hyperlipidemia in postmenopausal women.
Asunto(s)
Medicamentos Herbarios Chinos/uso terapéutico , Estrógenos/farmacología , Hiperlipidemias/tratamiento farmacológico , Metabolismo de los Lípidos/efectos de los fármacos , Menopausia , Fitoestrógenos/farmacología , Animales , Neoplasias de la Mama/tratamiento farmacológico , Grosor Intima-Media Carotídeo , Colesterol/metabolismo , Proteínas Quinasas Dependientes de AMP Cíclico/metabolismo , Femenino , Células Hep G2/metabolismo , Humanos , Hidroximetilglutaril-CoA Reductasas/efectos de los fármacos , Inhibidores de Hidroximetilglutaril-CoA Reductasas/farmacología , Hiperlipidemias/etiología , Lípidos/sangre , Lipoproteínas LDL/metabolismo , Células MCF-7/metabolismo , Modelos Animales , Fosforilación/efectos de los fármacos , Ratas , Ratas Sprague-Dawley , beta-CiclodextrinasRESUMEN
The regeneration of mitochondria by regulated biogenesis plays an important homeostatic role in cells and tissues and furthermore may provide an adaptive mechanism in certain diseases such as sepsis. The heme oxygenase (HO-1)/carbon monoxide (CO) system is an inducible cytoprotective mechanism in mammalian cells. Natural antioxidants can provide therapeutic benefit, in part, by inducing the HO-1/CO system. This study focused on the mechanism by which the natural antioxidant quercetin can induce mitochondrial biogenesis in HepG2 cells. We found that quercetin treatment induced expression of mitochondrial biogenesis activators (PGC-1 α , NRF-1, TFAM), mitochondrial DNA (mtDNA), and proteins (COX IV) in HepG2 cells. The HO inhibitor SnPP and the CO scavenger hemoglobin reversed the effects of quercetin on mitochondrial biogenesis in HepG2 cells. The stimulatory effects of quercetin on mitochondrial biogenesis could be recapitulated in vivo in liver tissue and antagonized by SnPP. Finally, quercetin conferred an anti-inflammatory effect in the liver of mice treated with LPS and prevented impairment of mitochondrial biogenesis by LPS in vivo. These salutary effects of quercetin in vivo were also antagonized by SnPP. Thus, our results suggest that quercetin enhances mitochondrial biogenesis mainly via the HO-1/CO system in vitro and in vivo. The beneficial effects of quercetin may provide a therapeutic basis in inflammatory diseases and sepsis.
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Hemo-Oxigenasa 1/metabolismo , Mitocondrias/enzimología , Recambio Mitocondrial/efectos de los fármacos , Quercetina/farmacología , Animales , Monóxido de Carbono/metabolismo , Activación Enzimática/efectos de los fármacos , Células Hep G2 , Humanos , Lipopolisacáridos/farmacología , Masculino , Ratones , Ratones Endogámicos C57BL , Mitocondrias/efectos de los fármacos , Proteínas Mitocondriales/metabolismo , Factores de TiempoRESUMEN
Seaweed extracts show diverse bioactivities, such as antioxidant and antitumor activity. Capsosiphon fulvescens is a green alga that is abundant along the southwest coast of South Korea. Although it is consumed for its purported health-enhancing properties, particularly as a treatment for stomach disorders and hangovers, the health effects of dietary C. fulvescens remain unclear. We extracted polysaccharides from C. fulvescens (Cf-PS), investigated their effects on the proliferation of rat small intestinal epithelial IEC-6 cells, and determined the signaling cascade involved. We cultured IEC-6 cells in the presence of Cf-PS, which stimulated cell proliferation in a dose-dependent manner, and analyzed the Wnt and MAPK signaling pathways, which are related to cell proliferation. Cf-PS treatment induced the translocation of ß-catenin, an effector of the Wnt signaling pathway, from the cytosol to the nucleus and increased the expression of cyclinD1 and c-myc. Cf-PS also induced ERK1/2 phosphorylation, which is activated by mitogenic and proliferative stimuli such as growth factors, but the phosphorylation of JNK and p38 was not enhanced. Our results show that Cf-PS regulates proliferation via stimulating the nuclear translocation of ß-catenin and ERK1/2 activation in intestinal epithelial cells.
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Proliferación Celular/efectos de los fármacos , Chlorophyta/química , Proteínas Quinasas Activadas por Mitógenos/metabolismo , Extractos Vegetales/farmacología , Polisacáridos/farmacología , Transducción de Señal/efectos de los fármacos , Análisis de Varianza , Animales , Western Blotting , Línea Celular , Relación Dosis-Respuesta a Droga , Electroforesis en Gel de Agar , Microscopía , Extractos Vegetales/aislamiento & purificación , Polisacáridos/aislamiento & purificación , Ratas , República de Corea , beta Catenina/metabolismoRESUMEN
We isolated a novel glycoprotein from the brown alga Laminaria japonica that has antiproliferative effects on HT-29 colon cancer cells. We also identified the mechanism by which this glycoprotein, named LJGP, induces apoptosis. MTS assays showed that LJGP inhibited the proliferation of several cancer cell lines (AGS, HepG2, HT-29) in a dose-dependent manner. Especially in HT-29 cells, proliferation was significantly decreased. LJGP treatment on HT-29 displayed several apoptotic features, such as DNA fragmentation, sub-G1 arrest, caspase-3 activation, and PARP degradation. Consistent with sub-G1 arrest, LJGP decreased the expression of Cdk2, cyclin E, cyclin D1, PCNA, E2F-1, and phosphorylated pRb. Furthermore, the increase of p27 expression was observed. We also determined that LJGP-induced apoptosis leads to the formation of a death-induced signaling complex of Fas, FADD, and procaspase-8. LJGP induced the reduction of mitochondrial membrane potential with activation of the Bcl-2 family of proteins and caspase-9. These findings suggest that LJGP inhibits HT-29 cell proliferation by inducing apoptosis, which may be mediated via multiple pathways, including the Fas signaling pathway, the mitochondrial pathway, and cell cycle arrest. Therefore, LJGP can be a useful treatment option for colon cancer in humans.
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Antineoplásicos/farmacología , Apoptosis/efectos de los fármacos , Neoplasias del Colon/tratamiento farmacológico , Glicoproteínas/farmacología , Laminaria/química , Antineoplásicos/análisis , Ciclo Celular/efectos de los fármacos , Ciclo Celular/fisiología , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Glicoproteínas/análisis , Células HT29 , Humanos , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Extractos Vegetales/farmacología , Ensayo de Tumor de Célula MadreRESUMEN
The brown alga Laminaria japonica is frequently consumed in Korea, Japan and China, and has been used for more than a thousand years as a drug in traditional Chinese medicine. In this study, we isolated a novel glycoprotein from L. japonica that stimulates the growth of the IEC-6 normal murine intestinal epithelial cells. We also identified the mechanism by which this glycoprotein, referred to as LJGP, stimulates cell growth. After 24 h of exposure to LJGP, cell proliferation increased in a dose-dependent manner. To further explore the mechanism associated with LJGP-induced cell proliferation, we treated cells for various times with LJGP. We focused on the epidermal growth factor receptor (EGFR) signaling pathway, which is involved in the regulation of cellular proliferation and differentiation, during LJGP-induced cell growth. The results showed that LJGP induced EGFR and Akt activation. Furthermore, LJGP stimulated Shc/Grb2 binding and ERK activation, but inhibited JNK phosphorylation. These results indicate that LJGP stimulates gastrointestinal cell growth by activating the EGFR signaling pathway.