RESUMEN

Orostachys japonicus is a popular traditional medicinal herb used in Asian countries. This study is focused on evaluating its role in lipid and glucose metabolism in cell and animal models to establish the plant as an anti-obesity and antidiabetic herb. A butanol fraction of O. japonicus was used in the study. The lipid production was evaluated by the Oil Red O technique while the expression of adipogenic markers by Western blotting and RT-PCR using 3T3-L1 preadipocyte. The effect on glucose uptake activity was evaluated in C2C12 myoblast cells. The animal study was carried out in C57BL mice to evaluate anti-obesity activity using the high-fat diet model. The evaluation of serum lipid, blood glucose, adipogenic and fibrosis markers in the liver, and fat deposition in the liver and adipose tissue (by histology) of mice was conducted. Butanol fraction of O. japonicus significantly inhibited the lipid production in the 3T3-L1 cells and reduced the expression of PPARγ, C/EBPα, SREBP-1c and aP2. It enhanced glucose uptake in insulin-resistant C2C12 myoblast cells. It reduced body weight, triglycerides, and blood glucose in the obese mice. It significantly inhibited lipid accumulation in the liver and adipose tissue of obese mice along with suppression of expression of adipogenic and fibrosis markers in the liver. In summary, supporting the previous results, this study helped to establish the potent anti-obesity, antidiabetic, and liver-protecting effect of the butanol fraction of O. japonicus.

RESUMEN

Acne vulgaris is a common skin disease characterized by increased sebum production, inflammation, and Cutibacterium acnes (CA: formerly Propionibacterium acnes) hyperproliferation in pilosebaceous follicles. This study evaluated the efficacy of FRO, a formula composed of fermented Rhus verniciflua Stokes and Orostachys japonicus, against acne pathogenesis via antimicrobial assessment and an in vitro analysis. Stimulated model cells treated with hormones, CA, or lipopolysaccharide (LPS) were designed based on the characteristics of acne pathogenesis, including inflammation and sebum hypersecretion. High-performance liquid chromatography, disc diffusion, MTS, and western blotting assays were used to examine potential anti-acne effects. FRO was determined to contain phenolics such as gallic acid, fisetin, quercetin, and kaempferol. FRO exerted antimicrobial activity against CA and inhibited reactive oxygen species production that was otherwise increased by LPS or CA in HaCaT cells. Additionally, FRO exerted anti-inflammatory effects by inhibiting iNOS, TNF-α, IL-6, p-STAT-3, and p-NF-κB, which were previously upregulated by LPS or CA in THP-1 and HaCaT cells. FRO inhibited lipogenesis induced by steroid hormones and CA by decreasing FAS and SREBP-1 levels in sebocytes. Additionally, FRO down-regulated the androgen receptor, 5α-reductase, SREBP-1, and FAS levels, which were upregulated by steroid hormone in LNCaP cells. Taken together, our findings suggest that FRO alleviates acne by inhibiting the growth of CA, inflammation, and excess sebum and could be used for functional cosmetics or acne treatments.

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The risk of atopic dermatitis (AD)-like skin lesions has increased due to the elevated levels of allergens worldwide. Natural-origin agents, which are effective and safe, show promise for the prevention and treatment of inflammatory conditions. Orostachys japonicus (OJ) A. Berger is an ingredient of traditional herbal medicines for fever, gingivitis, and cancer in Korea, China, and Japan. However, the effect of OJ on AD-like skin lesions is unknown. Therefore, we investigated the effect of OJ ethanol extract (OJEE) on AD-like skin symptoms in mice and cells. OJEE reduced the 2,4-dinitrochlorobenzene-induced AD severity, serum levels of IgE and TARC, and mRNA levels of TARC, TNF-α, and IL-4 in NC/Nga mice. Histopathological analysis showed that OJEE reduced the thickness of the epidermis/dermis and dermal infiltration of inflammatory cells in ear tissue. Furthermore, OJEE suppressed the TNF-α/IFN-γ-increased TARC mRNA level by inhibiting NF-κB and STAT1 activation in HaCaT cells. Taken together, our findings show that OJEE reduced the risk of AD-like skin symptoms by decreasing TARC expression via inhibiting NF-κB and STAT1 activation in skin keratinocytes and thus shows promise as an alternative therapy for AD-like skin lesions.

RESUMEN

Orostachys japonicus (O. japonicus) was extracted with ethanol (EtOH) and sequentially separated with organic solvents, including n-hexane (Hex), dichloromethane (DCM), ethyl acetate (EtOAc), n-butanol (BuOH), and water (H2O). All the fractions were confirmed for anti-inflammatory activity in an inflammatory condition. The DCM fraction showed the highest anti-inflammatory ability. Here, we examined the effect of DCM fraction and investigated the intracellular signaling pathways in LPS-stimulated RAW 264.7 macrophage cells. The DCM fraction significantly inhibited the mRNA levels of pro-inflammatory mediators and cytokines including iNOS, COX-2, IL-1ß, IL-2, IL-6, and IP-10 in LPS-stimulated cells. Also, the treatment of DCM fraction excellently reduced the expression of the proteins of AP-1 (phospho-c-Jun and phospho-c-Fos) and phospho-IRF3 as transcription factors. As a result, it suppressed LPS-induced inflammatory mediator and cytokines via inhibition of transcription factors. In conclusion, our data demonstrated that DCM fraction has a strong anti-inflammatory activity that improves the inflammatory state.

RESUMEN

ETHNOPHARMACOLOGICAL RELEVANCE: Orostachys japonicus A. Berger (O. japonicus), so-called Wa-song in Korea, a traditional food and medicine that grows on mountain rocks and roof tiles. Wa-song containing various phenolic compounds have been reported as a medicinal plant for prevention of fibrosis, cancer, inflammation, and oxidative damage. AIM OF THE STUDY: The present study was designed to examine the anti-angiogenic effects of cultivated Orostachys japonicus 70% ethanol extract (CE) in vascular endothelial growth factor (VEGF)-stimulated human umbilical vein endothelial cells (HUVECs). MATERIALS AND METHODS: CE was prepared with 70% ethanol. HUVECs, rat aortic rings, and matrigel plug in mice were treated with CE (10-20 µg/mL) and VEGF (20-50 ng/mL), and the anti-angiogenic activities of CE were analyzed by SRB, wound healing, trans-well invasion, capillary-like tubule formation, rat aortas, Western blot, and matrigel plug assay. Phenolic compounds in CE were analyzed using a high-performance liquid chromatography (HPLC)-PDA system. RESULTS: Treatment of CE (10-20 µg/mL) markedly suppressed proliferation of HUVECs in the presence (from 136.5% to 112.2%) or absence of VEGF (from 100.0% to 92.1%). The proliferation inhibitory effect of CE was caused by G0/G1 cell cycle arrest, and the decrease of CDK-2, CDK-4, Cyclin D1 and Cyclin E1. Furthermore, CE treatment showed significant angiogenesis inhibitory effects on motility, invasion and micro-vessel formation of HUVECs, rat aortic rings and subcutaneous matrigels under VEGF-stimulation condition. In HUVECs, CE-induced anti-angiogenic effect was regulated by inhibition of the PI3K/AKT/mTOR, MAPK/p38, MAPK/ERK, FAK-Src, and VEGF-VEGFR2 signaling pathways. CONCLUSION: This study demonstrated that CE might be used as a potential natural substance, multi-targeted angiogenesis inhibitor, functional food material.

Asunto(s)

Inhibidores de la Angiogénesis/farmacología , Crassulaceae/química , Neovascularización Patológica/tratamiento farmacológico , Extractos Vegetales/farmacología , Transducción de Señal/efectos de los fármacos , Factor A de Crecimiento Endotelial Vascular/metabolismo , Receptor 2 de Factores de Crecimiento Endotelial Vascular/metabolismo , Inductores de la Angiogénesis/farmacología , Animales , Puntos de Control del Ciclo Celular/efectos de los fármacos , Línea Celular , Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Colágeno/efectos de los fármacos , Colágeno/metabolismo , Combinación de Medicamentos , Fase G1/efectos de los fármacos , Células Endoteliales de la Vena Umbilical Humana , Humanos , Laminina/efectos de los fármacos , Laminina/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Neovascularización Patológica/metabolismo , Proteoglicanos/efectos de los fármacos , Proteoglicanos/metabolismo , Ratas , Ratas Sprague-Dawley , Fase de Descanso del Ciclo Celular/efectos de los fármacos

RESUMEN

Objective: To determine the bioactive compounds of Orostachys japonicus (O. japonicus) (Maxim.) A. Berger extraction by different solvents (70％ ethanol or water) and to evaluate the in vitro antioxidant and anti-inflammatory activities.Methods: Total polyphenol, flavonoid, and anthocyanin contents in O. japonicus extract were measured. Antioxidant activities were determined by DPPH, ABTS, and superoxide radical-scavenging ability assays. Anti-inflammatory activities were evaluated by nitric oxide production, tumor necrosis factor-α, and interleukin-6 expression techniques. Results: Extraction with 70％ ethanol yielded the highest total polyphenol (60.03 mg/g dry weight) and flavonoid (55.50 mg/g dry weight) contents. The total anthocyanin contents in 70％ ethanol and water extracts were 57.25 and 91.71 mg/g dry weight, respectively. The 70％ ethanol extract also showed stronger antioxidant activity than the water extract. Antioxidant activity and reducing power increased with the increasing concentration of O. japonicus extract. O. japonicus extract at 0-400 μg/mL did not affect the growth of RAW 264.7 cells, whereas dose-dependent inhibition of nitric oxide, tumor necrosis factor-α, and interleukin-6 production was observed. Conclusions: O. japonicus inhibits oxidative and inflammatory reactions with potentially positive health-related effects.

RESUMEN

To investigate the effect of Orostachys (O.) japonicus, a perennial herbaceous plant of the Family Crassulaceae, on biofilm formed by methicillin-resistant Staphylococcus aureus (MRSA). Methods: Powdered O. japonicus was extracted by 95% methanol, concentrated, and then, systematically fractionated with n-heane, dichloromethane (DCM), ethyl acetate (EtOAc), n-butanol, and H2O according to polarity. Among them, the flavonoid-rich EtOAc fraction demonstrated the highest antibacterial activity and was used in this study. Using the biofilm inhibition assay, cell-surface attachment assay, confocal laser scanning microscopy, latex agglutination assay, and real time qRT-PCR, we examined whether the EtOAc fraction inhibited the formation of MRSA biofilm. Results: The EtOAc fraction exhibited distinct activity against biofilm formation and cell-surface attachment of MRSA up to 1 mg/mL through down-regulating the expression of mecA gene and the production and agglutination of penicillin-binding protein 2a as solidly observed in biofilm inhibition assay, cell-suface attachment assay, confocal laser scanning microscopy, latex agglutination assay, and real time qRT-PCR analysis. Conclusions: These results suggest that O. japonicus could be utilized as a potential resource for the development of new antibiofilm formation of MRSA and antibacterial agents in the future.

RESUMEN

To determine the bioactive compounds of Orostachys japonicus (O. japonicus) (Maxim.) A. Berger extraction by different solvents (70% ethanol or water) and to evaluate the in vitro antioxidant and anti-inflammatory activities. Methods: Total polyphenol, flavonoid, and anthocyanin contents in O. japonicus extract were measured. Antioxidant activities were etermined by DPPH, ABTS, and superoxide radical-scavenging ability assays. Anti-inflammatory activities were evaluated by nitric oxide production, tumor necrosis factor-a, and interleukin-6 expression techniques. Results: Extraction with 70% ethanol yielded the highest total polyphenol (60.03 mg/g dry weight) and flavonoid (55.50 mg/g dry weight) contents. The total anthocyanin contents in 70% ethanol and water extracts were 57.25 and 91.71 mg/g dry weight, respectively. The 70% ethanol extract also showed stronger antioxidant activity than the water extract. Antioxidant activity and reducing power increased with the increasing concentration of O. japonicus extract. O. japonicus extract at 0-400 ug/mL did not affect the growth of RAW 264.7 cells, whereas dose-dependent inhibition of nitric oxide, tumor necrosis factor-a, and interleukin-6 production was observed. Conclusions: O. japonicus inhibits oxidative and inflammatory reactions with potentially positive health-related effects.

RESUMEN

To investigate the effect of Orostachys (O.) japonicus, a perennial herbaceous plant of the Family Crassulaceae, on biofilm formed by methicillin-resistant Staphylococcus aureus (MRSA). Methods: Powdered O. japonicus was extracted by 95% methanol, concentrated, and then, systematically fractionated with n-heane, dichloromethane (DCM), ethyl acetate (EtOAc), n-butanol, and H2O according to polarity. Among them, the flavonoid-rich EtOAc fraction demonstrated the highest antibacterial activity and was used in this study. Using the biofilm inhibition assay, cell-surface attachment assay, confocal laser scanning microscopy, latex agglutination assay, and real time qRT-PCR, we examined whether the EtOAc fraction inhibited the formation of MRSA biofilm. Results: The EtOAc fraction exhibited distinct activity against biofilm formation and cell-surface attachment of MRSA up to 1 mg/mL through down-regulating the expression of mecA gene and the production and agglutination of penicillin-binding protein 2a as solidly observed in biofilm inhibition assay, cell-suface attachment assay, confocal laser scanning microscopy, latex agglutination assay, and real time qRT-PCR analysis. Conclusions: These results suggest that O. japonicus could be utilized as a potential resource for the development of new antibiofilm formation of MRSA and antibacterial agents in the future.

RESUMEN

Targeted therapy at the molecular level is important for pancreatic cancer treatment. This study looked over the anticancer activity of Orostachys japonicus in a human pancreatic cancer cell line, PANC-1. An ethyl acetate fraction containing quercetin, kaempferol, and flavonol glycosides from O. japonicus (OJE) exhibited significant anticancer activity against the PANC-1. OJE activated caspase-3, caspase-8, and caspase-9, leading to the induction of both intrinsic and extrinsic apoptosis pathways. It also inhibited cyclin D1, cyclin B1, and cyclin-dependent kinase 4, representing cell cycle arrest at both G1/S and G2/M phases. In addition, OJE phosphorylated MAPKs such as p38, JNK, and ERK, which are important upstream signaling factors in apoptosis and arrest of cell cycle inducing system. In conclusion, OJE effectively exerted antipancreatic cancer activity via induction of apoptosis directed by both intrinsic and extrinsic pathways and arrest of cell cycle regulated at both G1/S and G2/M stages, which is activated by MAPKs, p38, JNK, and ERK.

RESUMEN

In this study, Orostachys japonicus was extracted with ethyl alcohol and fractionated by a serial of organic solvents. The ethyl acetate fraction was found to be the most effective among the tested five fractions. High-performance liquid chromatography and mass spectrometry analysis of the ethyl acetate fraction presented epicatechin gallate, quercetin-3-O-glucoside, and kaempferol-3-O-rutinoside. Treatment with O. japonicus inhibited reactive oxygen species (ROS) generation and lipid accumulation during adipogenesis. The gene expression of enzymes involved in the antioxidant system increased in O. japonicus-treated cells. messeanger RNA (mRNA) and protein expression of the pro-oxidant enzymes such as nicotinamide adenine dinucleotide phosphate hydrogen oxidase4 and glucose-6-phosphate dehydrogenase suppressed in O. japonicus-treated cells. O. japonicus also inhibited the mRNA and protein levels of adipogenic transcription factors (including proliferator activated receptor-γ and CCAAT/enhancer-binding protein-α) and their target gene (adipocyte protein 2). These results suggest that O. japonicus inhibits adipogenesis by controlling pro-/anti-oxidant enzyme responses and adipogenic transcription factors. PRACTICAL APPLICATIONS: ROS generation is markedly related to the pathogenesis and development of metabolic disorders. Treatment with O. japonicus inhibited ROS generation and lipid accumulation during adipogenesis. This result indicates that O. japonicus inhibit adipogenesis by controlling pro-/anti-oxidant enzyme responses and adipogenic mediators.

Asunto(s)

Crassulaceae/química , Metabolismo de los Lípidos/efectos de los fármacos , Fenoles/química , Fenoles/farmacología , Extractos Vegetales/farmacología , Células 3T3-L1 , Acetatos , Adipogénesis/efectos de los fármacos , Adipogénesis/fisiología , Animales , Diferenciación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Depuradores de Radicales Libres/metabolismo , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Ratones , Capacidad de Absorbancia de Radicales de Oxígeno , Extractos Vegetales/química , Especies Reactivas de Oxígeno

RESUMEN

Orostachys japonicus has traditionally been used as a food product and a fork medicine in Asia to treat various diseases. Angiogenesis is a critical process that contributes to various chronic diseases via excessive delivery of oxygen and nutrients. Common anti-angiogenic drugs have serious problems related to high costs and side effects; thus, natural products with low costs and no cytotoxicity have garnered increasing interest. In this study, we evaluated and compared the anti-angiogenic effects and phenolic compound contents between wild (WOEs) and cultivated O. japonicus extracts (COEs) prepared under various extract conditions. WOEs and COEs suppressed cell proliferation of human umbilical vein endothelial cells (HUVECs) and inhibited vascular endothelial growth factor-induced chemotactic migration, invasion, and capillary-like tube formation in HUVECs. Among COEs, that prepared by 70% EtOH (70% CE) showed the most effective anti-angiogenic activity in HUVECs. When compared to WOEs, total polyphenol and total flavonoid contents were 1.28 to 4.38 times higher in COEs, and 70% CE contained the greatest flavonoid contents (28.28 ± 0.93 mg%), as well as the highest levels of major phenolic compounds including gallic acid (21.84 µg/mL), epicatechin-gallate (6.58 µg/mL), kaempferol (6.32 µg/mL), and quercetin (8.55 µg/mL). Although further studies are required to identify the molecular mechanisms behind these anti-angiogenic effects, 70% CE could be used as an herbal medicine, functional food ingredient, and potent angiogenesis inhibitor. PRACTICAL APPLICATION: Environmental factors such as altitude, nutrients, exposure to sunlight, and temperature can influence the type and quantity of bioactive components in plants. The advantage of cultivated plants is that the above-mentioned factors can be artificially adjusted compared to wild plants. Based on economic efficiency, productivity, and consistent quality, anti-angiogenesis activity of cultivated O. japonicus is of greater commercial value as a functional food than wild O. japonicus.

Asunto(s)

Inhibidores de la Angiogénesis/farmacología , Crassulaceae/química , Células Endoteliales de la Vena Umbilical Humana/efectos de los fármacos , Extractos Vegetales/farmacología , Plantas Medicinales/química , Inhibidores de la Angiogénesis/química , Animales , Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Crassulaceae/crecimiento & desarrollo , Flavonoides/química , Flavonoides/farmacología , Ácido Gálico/química , Ácido Gálico/farmacología , Células Endoteliales de la Vena Umbilical Humana/citología , Células Endoteliales de la Vena Umbilical Humana/metabolismo , Humanos , Fenoles/química , Fenoles/farmacología , Extractos Vegetales/química , Plantas Medicinales/crecimiento & desarrollo , Factor A de Crecimiento Endotelial Vascular/metabolismo

RESUMEN

Orostachys japonicus (O. japonicus) is utilized as a traditional medicine for patients with various diseases. This study investigated the effect of the ethyl acetate fraction from O. japonicus extract (OJE) on the growth inhibition of OVCAR-3 human ovarian cancer cells demonstrated to inhibit cell growth and arrest the cell cycle in OVCAR-3 cells by blocking the sub-G1 phase and decreasing cyclin E1/CDK2 expression. Cell cycle arrest was connected to the increased expression of the cell cycle regulating factors p53 and p21. Apoptosis was initiated through the intrinsic pathway by up-regulating the expression of the Bcl-2/Bax ratio and down-regulating the expression of pro-caspase-3. Furthermore, OJE treatment elicited p-p38 activation and p-ERK1/2 inhibition. In conclusion, our results demonstrated that OJE reduced the growth of OVCAR-3 human ovarian cancer cells mediated by arrest of the cell cycle and regulation of MAPK signaling pathways.

RESUMEN

Objective:To investigate the anti-colon cancer effects of ethylacetate fraction from Orostachys japonicus (O. japonicus) on HT-29 cancer cells.Methods:The viability of HT-29 cells was assayed by the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) method. Apoptosis induction and cell cycle inhibition were confirmed by fluorescein isothiocyanate and propidium iodide staining using flow cytometry. Morphological changes in the nucleus were observed, using a fluorescence microscope with 4′,6-diamidino-2-phenylindole (DAPI) nuclear staining. The expression levels of the upstream and downstream proteins involved in the anti-cancer mechanism were confirmed by Western blotting.Results:After treating HT-29 cells with different concentrations of ethylacetate fraction from O. japonicus, the viability of cells decreased in a concentration-dependent manner, while apoptosis induction and apoptotic body formation increased. Cell cycle analysis showed that the arrest occurred at the sub-GConclusions:Combining the above results, it is thought that the survival of HT-29 cells is suppressed by ethylacetate fraction from O. japonicus through mitochondrial regulation-induced caspase cascade activation, induction of apoptosis and cell cycle arrest.

RESUMEN

Objective: To investigate the anti-colon cancer effects of ethylacetate fraction from Orostachys japonicus (O. japonicus) on HT-29 cancer cells. Methods: The viability of HT-29 cells was assayed by the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) method. Apoptosis induction and cell cycle inhibition were confirmed by fluorescein isothiocyanate and propidium iodide staining using flow cytometry. Morphological changes in the nucleus were observed, using a fluorescence microscope with 4′,6-diamidino-2-phenylindole (DAPI) nuclear staining. The expression levels of the upstream and downstream proteins involved in the anti-cancer mechanism were confirmed by Western blotting. Results: After treating HT-29 cells with different concentrations of ethylacetate fraction from O. japonicus, the viability of cells decreased in a concentration-dependent manner, while apoptosis induction and apoptotic body formation increased. Cell cycle analysis showed that the arrest occurred at the sub-G

RESUMEN

Orostachys japonicus A. Berger (), known as Wa-song in Korea, has been reported to exert various biological effects, such as anti-tumor, anti-oxidant, and anti-febrile effects. However, the anti-angiogenic effects of O.japonicus extracts remain to be investigated. In the present study, we demonstrated the anti-angiogenic effects of bioconverted O. japonicus extract (BOE) in Ms-1 mouse endothelial cells and compared them with the bioactivities of O. japonicus extract (OE). BOE, but not OE, were found to exert anti-angiogenic effects, including inhibition of cell migration, cell adhesion, tube formation of Ms-1 cells, and blood vessel formation of matrigel plug assay in vivo. Furthermore, protein levels of phosphorylated Src kinase were lower in BOE-treated cells than in OE-treated cells. Treatment with OE or BOE did not influence cell viability during the experimental period. Bioconverted extract of O.japonicus have anti-angiogenic effects in vitro and vivo, but non-bioconverted extract do not. We suggest that these observed anti-angiogenic effects are caused by the changes in the composition of bioactive compounds in the extracts as a result of biological conversion.

Asunto(s)

Crassulaceae/química , Células Endoteliales/citología , Extractos Vegetales/química , Extractos Vegetales/farmacología , Inhibidores de la Angiogénesis/química , Inhibidores de la Angiogénesis/farmacología , Animales , Western Blotting , Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Células Endoteliales/efectos de los fármacos , Masculino , Ratones , Ratones Endogámicos C57BL , Ensayo de Radioinmunoprecipitación

RESUMEN

Orostachys japonicus is an herb that contains several functional components and has traditionally been used to treat various diseases in Asia. In this study, bioactive components from different parts of the O. japonicus plant were investigated, and the contents of the functional components in ethanol extracts of O. japonicus cultivated in Korea and China were compared. The antioxidant effects of O. japonicus ethanol extracts were investigated using Raw 264.7 cells. It was found that 2,2-diphenyl-1-picrylhydrazyl radical-scavenging activity was significantly decreased in the cells treated with the extracts. Moreover, the novel inhibitory functions of O. japonicus extracts on collagenase, elastase, and tyrosinase were established. We also found that O. japonicus extracts strongly inhibited melanin synthesis in B16F10 melanoma cells by decreasing MITF protein levels and activating the Erk and Akt signaling pathways. Thus, these findings would be useful for developing new cosmetic and pharmaceutical formulations based on O. japonicus extracts.

Asunto(s)

Colagenasas/metabolismo , Crassulaceae/química , Quinasas MAP Reguladas por Señal Extracelular/metabolismo , Extractos Vegetales/farmacología , Proteínas Proto-Oncogénicas c-akt/metabolismo , Animales , Línea Celular Tumoral , China , Etanol/química , Melaninas/antagonistas & inhibidores , Melaninas/biosíntesis , Melanoma Experimental/metabolismo , Melanoma Experimental/patología , Ratones , Monofenol Monooxigenasa/antagonistas & inhibidores , Monofenol Monooxigenasa/metabolismo , Elastasa Pancreática/antagonistas & inhibidores , Elastasa Pancreática/metabolismo , Extractos Vegetales/química , Células RAW 264.7 , República de Corea , Transducción de Señal/efectos de los fármacos

RESUMEN

Cholinesterase inhibitors block the bioconversion of neurotransmitters by cholinesterase in the nervous system. Epicatechin derivatives (1, 3 and 5), polyphenols (6 and 7) from Orostachys japonicus, and catechin derivatives (2 and 4) from our in-house library were evaluated for their inhibitory activity on cholinesterase. Compound 5 exhibited IC50 values of 58.3±2.4 and 17.8±3.8µg/mL on AChE and BuChE, respectively. Compound 5 inhibited BuChE more strongly than AChE through a competitive behavior. In silico binding positions of 5 in the active site were predicted using Autodock 4.2 and processed in a 10000-ps molecular dynamics simulation to assess the stability of compound 5 binding.

Asunto(s)

Butirilcolinesterasa/metabolismo , Catequina/análogos & derivados , Catequina/farmacología , Inhibidores de la Colinesterasa/química , Inhibidores de la Colinesterasa/farmacología , Crassulaceae/química , Antioxidantes/química , Antioxidantes/metabolismo , Antioxidantes/farmacología , Butirilcolinesterasa/química , Catequina/metabolismo , Inhibidores de la Colinesterasa/metabolismo , Humanos , Cinética , Simulación del Acoplamiento Molecular , Simulación de Dinámica Molecular , Conformación Proteica , Especies Reactivas de Oxígeno/metabolismo

RESUMEN

ETHNOPHARMACOLOGICAL RELEVANCE: Orostachys japonicus A. Berger (Crassulaceae) (OJ), well-known as Wa-song in Korea is a medicinal plant with immunoregulatory, anti-febrile, antidote, and anti-cancer activities. This study was aimed at evaluating the immunostimulatory effect of O. japonicus A. Berger and its possible mechanisms of action. MATERIALS AND METHODS: To evaluate the effect of OJ aqueous extract on macrophage activity, we evaluated the modulation of macrophage activation state by observing structural (phagocytic activities) and the production of nitric oxide increase. The effect of OJ aqueous extract on RAW264.7 cell viability were assessed using Cell Counting Kit (CCK)-8 assay. HPLC analysis was performed to identify potential active compounds of this extract. RESULTS: The biological investigations indicated that OJ aqueous extract, among others, possessed the highest macrophage activation as indicated by NO production yield. The results showed that OJ aqueous extract exhibited antioxidant effects, which included scavenging activities against DPPH radicals. OJ aqueous extract increased the phagocytic activity of RAW 264.7 cells against IgG-opsonized red blood cells (RBC). The level of phosphorylated Syk kinase was increased in OJ aqueous extract-treated group as compared to control. Phosphorylation of PLC-γ was increased in the OJ aqueous extract-treated groups. Quercetin-3-O-rhamnose and kaempferol-3-O-rhamnose was detected in OJ aqueous extract by HPLC analysis. CONCLUSIONS: OJ aqueous extract might play a pivotal ethnopharmacologic role as an immunostimulatory agent by promoting Fc gamma receptor (FcγR)-mediated phagocytosis of IgG-opsonized RBCs. On the basis of our results, OJ aqueous extract can enhance innate immunity and may serve as an adjuvant for tumor treatment.

Asunto(s)

Adyuvantes Inmunológicos/farmacología , Crassulaceae/química , Macrófagos/efectos de los fármacos , Extractos Vegetales/farmacología , Adyuvantes Inmunológicos/aislamiento & purificación , Animales , Antioxidantes/aislamiento & purificación , Antioxidantes/farmacología , Línea Celular , Supervivencia Celular/efectos de los fármacos , Cromatografía Líquida de Alta Presión/métodos , Eritrocitos/efectos de los fármacos , Eritrocitos/metabolismo , Depuradores de Radicales Libres/aislamiento & purificación , Depuradores de Radicales Libres/farmacología , Inmunidad Innata/efectos de los fármacos , Activación de Macrófagos/efectos de los fármacos , Macrófagos/inmunología , Ratones , Óxido Nítrico/metabolismo , Fagocitosis/efectos de los fármacos , Ovinos

RESUMEN

The hypolipidemic and hypoglycemic effects of two dietary dosages (0.1% and 0.5%) of water and 80% ethanol extracts from hot-air dried Orostachys japonicus A. Berger were evaluated in the serum and organ tissues of streptozotocin-induced diabetic rats. The STZ-induced diabetic groups supplemented with the O. japonicus extracts showed significantly higher body weight compared to a diabetic control group at the end of experiment. The extracts exhibited substantial hypoglycemic effects by significant reductions of fasting blood glucose levels at all time points tested compared to the initial stage before treatment of the extracts. Declines of serum and hepatic triglyceride levels were greater than declines of total cholesterol in the groups treated with the 0.5% O. japonicus extract (DBW2 and DBE2) when compared to the DBC group. Hepatic glycogen content was higher in the groups treated with O. japonicus extract, while lipid peroxide content was decreased in these treated groups compared to the DBC group. Hepatic antioxidant activity was significantly increased in the groups supplemented with the O. japonicus ethanol extract. The hypolipidemic and hypoglycemic effects of the O. japonicus ethanol extract were significantly greater than the effects of the water extract. Based on this study, it seems that O. japonicus ethanol extract, due to its higher phenolic and flavonoid components than the water extract, may control blood glucose and alleviate hyperlipidemia in diabetes.