Avaliação das respostas de biomineralização pelo Rubus coreanus frente à deficiência de estrógeno: um estudo translacional / Evaluation of biomineralization responses by Rubus coreanus to estrogen deficiency: a translational study

Caracterização da ação local do Rubus coreanus através da funcionalização da superfície dos implantes a serem instalados. Realizado em 2 etapas. Na primeira etapa, foi realizada a caracterização da superfície de implantes funcionalizadas pelos fármacos, a partir de teste físico químico e testes biológicos em culturas de células. Para a cultura de células foram utilizadas células mesenquimais indiferenciadas, isoladas de fêmures de ratas. Foram avaliadas a atividade e a diferenciação celular. Posteriormente, foi realizada a segunda etapa onde foram executados os experimentos in vivo. Para tanto, foram utilizadas 80 ratas Wistar adultas jovens, fêmeas, sendo 32 ratas para o primeiro período de eutanásia (14 dias), e 48 ratas para o segundo período de eutanásia (28 dias). Os grupos experimentais são: SHAM CONV, SHAM DMSO, SHAM RC 200, SHAM RC 400, OVX CONV, OVX DMSO, OVX RC 200 e OVX RC 400, sendo que os animais OVX foram submetidos a ovariectomia bilateral, enquanto que os animais SHAM foram submetidos à cirurgia fictícia de ovariectomia bilateral. Passados 30 dias, todos animais foram submetidos à cirurgia de instalação dos implantes nas metáfises tibiais. Os implantes tiveram as superfícies tratadas com DMSO ou RC 200 ou RC 400, e houve o controle negativo realizado através dos implantes convencionais. A eutanásia dos animais do primeiro período de eutanásia aconteceu aos 14 dias após a instalação dos implantes e as tíbias foram destinadas para a análise biomecânica (contra-torque). Para os animais do segundo período de eutanásia, aos 28 dias após a instalação dos implantes foi realizada a eutanásia dos animais, e as tíbias foram destinadas para as seguintes análises: biomecânica (contra-torque), enquanto do lado contralateral foi realizada a microtomografia computadorizada para avaliação morfométrica do osso formado ao redor dos implantes instalados. Os dados quantitativos foram submetidos ao teste de homocedasticidade para a seleção do teste estatístico apropriado (paramétrico ou não paramétrico), com nível de significância de 5%. Através das análises biomecânica e microtomográfica foi possível verificar resultados semelhantes entre os grupos experimentais. Contudo, o rubus coreanus não foi capaz de atuar de forma positiva no processo de reparo ósseo perimplantar(AU)

Characterization of the local action of Rubus coreanus through the functionalization of the surface of the implants to be installed. Carried out in 2 steps. In the first stage, the characterization of the surface of implants functionalized by drugs was carried out, based on physical-chemical tests and biological tests on cell cultures. For cell culture, undifferentiated mesenchymal cells, isolated from femurs of rats, were used. Cell activity and differentiation were evaluated. Subsequently, the second stage was carried out, where the in vivo experiments were performed. For that, 80 young adult female Wistar rats were used, 32 rats for the first period of euthanasia (14 days), and 48 rats for the second period of euthanasia (28 days). The experimental groups are: SHAM CONV, SHAM DMSO, SHAM RC 200, SHAM RC 400, OVX CONV, OVX DMSO, OVX RC 200 and OVX RC 400, with the OVX animals undergoing bilateral ovariectomy, while the SHAM animals underwent submitted to sham surgery of bilateral ovariectomy. After 30 days, all animals underwent surgery to install the implants in the tibial metaphysis. The implants had their surfaces treated with DMSO or RC 200 or RC 400, and there was a negative control performed using conventional implants. The animals from the first period of euthanasia were euthanized 14 days after implant placement and the tibiae were destined for biomechanical analysis (counter-torque). For animals in the second period of euthanasia, 28 days after implant placement, the animals were euthanized, and the tibiae were destined for the following analyses: biomechanics (counter-torque), while on the contralateral side, computerized microtomography was performed for morphometric evaluation of the bone formed around the installed implants. Quantitative data were submitted to the homoscedasticity test to select the appropriate statistical test (parametric or non-parametric), with a significance level of 5%. Through biomechanical and microtomographic analyzes it was possible to verify similar results between the experimental groups. However, rubus coreanus was not able to act positively in the peri-implant bone repair process(AU)

Estrogenic Activity of Sanguiin H-6 through Activation of Estrogen Receptor α Coactivator-binding Site

A popular approach for the study of estrogen receptor α inhibition is to investigate the protein-protein interaction between the estrogen receptor (ER) and the coactivator surface. In our study, we investigated phytochemicals from Rubus coreanus that were able to disrupt ERα and coactivator interaction with an ERα antagonist. The E-screen assay and molecular docking analysis were performed to evaluate the effects of the estrogenic activity of R. coreanus extract and its constituents on the MCF-7 human breast cancer cell line. At 100 µg/mL, R. coreanus extract significantly stimulated cell proliferation (574.57 ± 8.56%). Sanguiin H6, which was isolated from R. coreanus, demonstrated the strongest affinity for the ERα coactivator-binding site in molecular docking analysis, with a binding energy of

Anti-Obesity Effects on Unripe Rubus coreanus Miquel Extract in High Fat Diet-Induced Obese Mice.

Aims: The objective of the present study was to evaluate the anti-obesity effects of unripe Rubus coreanus Miquel (uRC) in 3T3-L1 adipocytes and body weight, epididymal fat and perirenal fat weight, and lipid profiles in diet-induced obese (DIO) C57BL/6 mice. Methodology: The lipid accumulation in 3T3-L1 adipocytes was carried out Oil Red O staining. And uRC (50 and 100 mg/kg/day) were orally administered for 90 days from the day of feeding with high fat diet (HFD). The serum total cholesterol (TC), triglyceride (TG), high density lipoprotein (HDL)-cholesterol and low density lipoprotein(LDL)-cholesterol and glucose levels were measured using Alere cholesterol LDXⓇ system. And the serum glutamic oxaloacetic transaminase (GOT), glutamic pyruvic transaminase (GPT), blood urea nitrogen(BUN) and creatinine levels were measured using the respective kits. Results: Our results indicated that treatment with uRC dose-dependently inhibited lipid accumulation in 3T3-L1 adipocytes. Moreover, after oral administration for 12 weeks, uRC (50 and 100mg/kg/day) extract produced a significant decrease in the serum total cholesterol (TC), lowdensity lipoprotein (LDL) cholesterol, glucose and glutamic-oxaloacetic transaminase (GOT) levels of HFD-induced obese mice. Similarly, uRC extract elevated serum high density lipoprotein (HDL) cholesterol. These results suggest that uRC extract may be a useful resource for the management of obesity. Conclusion: These results suggest that uRC extract may be a useful resource for the management of obesity.

Growth inhibition effect of Rubus coreanus Miquel on Candida albicans / 대한구강보건학회지

OBJECTIVES: In this study, the growth inhibition effect of Rubus coreanus Miquel on Candida albicans (C. albicans) was observed. METHODS: The Rubus coreanus Miquel was extracted with 70% methanol and concentrated with a rotary evaporator. Antifungal effect of Rubus coreanus Miquel extract on C. albicans was determined by paper disc diffusion method and standard plate count method. Seven different concentrations (2, 4, 8, 15, 30, 60, 120 mg/ml) of the extract were tested by paper disc diffusion method. Two kinds of concentration (8, 60 mg/ml) of the extract were tested using standard plate count method on C. albicans with different incubating time (for 6, 12, 24 hours immediately after the cultivation). Morphological changes of C. albicans cells after exposure to the extract were observed with scanning electron microscopy (SEM) images. RESULTS: The Rubus coreanus Miquel extract showed an antifungal effect on C. albicans in 8, 15, 30, 60, 120 mg/ml of concentrations (P<0.05). The extract with 8 mg/ml of concentration showed about 30% of growth inhibition at 6 h and with 60 mg/ml it showed about 90% of growth inhibition at 24 h. SEM analysis showed damaged surfaces of C. albicans cells when treated with Rubus coreanus Miquel extract. CONCLUSIONS: The Rubus coreanus Miquel might have the potential as a nobel growth inhibitory agent against C. albicans that causes oral infection.

Hepatoprotective effects of Rubus coreanus miquel concentrates on liver injuries induced by carbon tetrachloride in rats

As well-being foods pursuing healthy life are becoming popular, interest in Rubus coreanus Miquel (RCM) fruit, a type of Korean blackberry, is increasing due to its medicinal actions including protecting the liver, brightening the eyes, and alleviating diabetes. This study was carried out to evaluate the hepatoprotective effects of RCM concentrates on liver injuries induced by carbon tetrachloride (CCl4) in rats. RCM, produced in June ~ July 2008 at Chunbook, Gochang (South Korea), was finely mashed. The seeds were removed and the juices were condensed. Thirty-two Sprague-Dawley rats were divided into four groups according to treatment: normal (eight rats), CCl4, 1% RCM, and 2% RCM. Experimental diets were provided to the experimental animals for 4 weeks. We measure total cholesterol, high density lipoprotein-cholesterol (HDL-C), aspartate amino transferase (AST), alanine amino transferase (ALT), and alkaline phosphatase (ALP) levels. Part of the livers was isolated for histopathological evaluation, and analyzed for lipid peroxide (TBARS), superoxide dismutase (SOD) and liver proteins. The activities of serum AST, ALT, and ALP were elevated following CCl4 administration. Levels of hepatic TBARS were also significantly increased in the CCl4 groups. However, hepatic TBARS levels and the activities of serum enzymes were markedly reduced by supplementation with the RCM concentrates (P < 0.05). Hepatic SOD activity increased in the RCM concentrates group versus CCl4 groups. Histopathological examination revealed massive necrosis in the centrilobular area and degenerative changes caused by CCl4 were ameliorated by dietary supplementation with RCM concentrates. These results suggest that RCM concentrates have hepatoprotective effects and may improve the symptoms of liver injuries.

Anti-inflammatory effects of Rubus coreanus Miquel through inhibition of NF-kappaB and MAP Kinase

BACKGROUND/OBJECTIVES: Rubus Coreanus Miquel (RCM), used as a traditional Korean medicine, reduces chronic inflammatory diseases such as cancer and rheumatoid arthritis. However, its mechanism has not been elucidated. In this study, we examine the anti-inflammatory effects of RCM and their possible mechanisms using RAW 264.7 cells. MATERIALS/METHODS: Unripe RCM ethanol extract (UE), unripe RCM water extract (UH), ripe RCM ethanol extract (RE), and ripe RCM water extract (RH) were prepared. Inflammatory response was induced with LPS treatment, and expression of pro-inflammatory mediators (iNOS, COX-2, TNF-alpha, IL-1beta, and IL-6) and NO and PGE2 productions were assessed. To determine the anti-inflammatory mechanism of RCM, we measured NF-kappaB and MAPK activities. RESULTS: UE and UH treatment significantly reduced NF-kappaB activation and JNK and p38 phosphorylation and reduced transcriptional activities decreased iNOS, COX-2, and pro-inflammatory cytokines expressions, and NO and PGE2 productions. RE and RH treatments reduced IL-1beta and IL-6 expressions through suppressions of JNK and p38 phosphorylation. CONCLUSIONS: In this study, we showed that RCM had anti-inflammatory effects by suppression of pro-inflammatory mediator expressions. Especially, unripe RCM showed strong anti-inflammatory effects through suppression of NF-kappaB and MAPK activation. These findings suggest that unripe RCM might be used as a potential functional material to reduce chronic inflammatory responses.

Physiological Functionalities of Vitis hybrid (Sheridan)-Rubus coreanus Red Wine Made by Saccharomyces cerevisiae

Vitis hybrid (Sheridan)-Robus coreanus red wine was vinified by fermentation of a mixture of Vitis hybrid and Robus coreanus must at 25degrees C for 10 days. The Vitis hybrid-Robus coreanus red wine had ethanol contents of 10.9%. It had high antihypertensive angiotensin I-converting enzyme (ACE) inhibitory activity of 57.8% and antioxidant activity of 64.8%. Changes in the physicochemical properties and functionality of the Vitis hybrid-Robus coreanus red wine was investigated during a post-fermentation period of three months. The ACE inhibitory activity of the red wine increased as the post-fermentation period prolonged, and showed the highest ACE inhibitory activity of 70.4% 60 days post-fermentation. However, the antioxidant activity declined significantly to 47.2% during the post-fermentation period of 60 days. In terms of sensory evaluation, the Vitis hybrid-Robus coreanus red wine had the best acceptability 60 days post-fermentation.

Effects of a Rubus coreanus Miquel supplement on plasma antioxidant capacity in healthy Korean men

Korean raspberry, Rubus coreanus Miquel (RCM), contains high concentrations of phenolic compounds, which prevent oxidative stress. To determine the effect of RCM on antioxidant capacity in humans, we assessed in vivo lipid oxidation and antioxidant enzyme activities from plasma in 15 healthy men. The subjects ingested 30 g of freeze-dried RCM daily for 4 weeks. Blood was taken at baseline and at the end of the study to determine blood lipid profiles, fasting plasma glucose, liver function, lipid peroxidation, and antioxidant enzyme activities. RCM supplementation had no effect on blood lipid or fasting plasma glucose concentrations but decreased alkaline phosphatase activity. RCM supplementation increased glutathione peroxidase activities (P < 0.05) but had no effect on lipid peroxidation. These results suggest that short-term RCM supplementation may offer health benefits by enhancing antioxidant capacity in a healthy population.

A Study of Potential Application of Rubus coreanus Miquel Extract for Seborrheic Dermatitis Treatment / 대한의진균학회지

BACKGROUND: Seborrheic dermatitis is chronic relapsing inflammatory skin disorder. Bokbunja (Rubus coreanus Miquel) is a wild berry to Rosaceae genus and also known to have an anti-inflammation effect. OBJECTIVE: We were to determine the effect of Rubus coreanus Miquel extract for seborrheic dermatitis in vivo and in vitro. METHODS: Seven patients with mild seborrheic dermatitis were enrolled in this study. PCR and culture were performed to identify subtypes of six Malassezia species (M. restricta, M. globosa, M. furfur, M. slooffiae, M. sympodialis, M. obtusa). Topical application of Rubus coreanus Miquel Extract was applied twice daily for 2 weeks. Clinical improvement and safety assessment were performed initially and 2 weeks later. Minimum inhibitory concentration (MIC) was evaluated on Malassezia globosa comparing with ketoconazole and itraconazole. Sebum production was also checked prior the experiment and 2 weeks later. RESULTS: Five of seven patients showed improvement. No significant adverse effects were found during the clinical trial. Mild dryness was reported in 2 patients but they resolved spontaneously without any treatment. Rubus coreanus Miquel Extract didn't show antimicrobial effect to Malassezia globosa. However, Rubus coreanus Miquel Extract showed anti-inflammatory effect. CONCLUSION: In this study, we were verified that Rubus coreanus Miquel Extract can be applied for seborrheic dermatitis treatment. And this action mechanism is not related with antimicrobial effect.

Antioxidative and aldose reductase-inhibitory effects of a fermentation filtrate of Rubus coreanus / 한국실험동물학회지

Antioxidative and aldose reductase (AR)-inhibitory effects of a fermentation filtrate of Rubus coreanus (FRC) were investigated using corneal/retinal homogenate and lens cytosol, respectively. Rat corneal/retinal homogenate was treated with 50 microM FeCl3 in the presence of FRC (3.2-100 microg/mL) for 30 min at 37degrees C, and thiobarbituric acid-reactive substances (TBARS) was quantified as a lipid peroxidation parameter. FRC markedly suppressed the TBARS production in a concentration-dependent manner, leading to 50% (IC50) and 100% (IC100) inhibitory concentrations of 20 and 95 microg/mL, respectively, which was similar to the effect of butylated hydroxyanisole. Activity of AR from rat lens was assayed in the presence of FRC (1-31.6 microg/mL) at 25degrees C using glyceraldehyde as a substrate. FRC inhibited lens AR by 50% (IC50) and 90% (IC90) at approximately 2 and 31.6 microg/mL, respectively, comparable to the effect of quercetin. The results indicate that ERC could be a promising candidate for the improvement of eye injury and visual dysfunction of dry eye and diabetic patients.

Treatment for Two Cases of Acne Vulgaris with Rubus Coreanus Miquel Extract / 대한의진균학회지

Bokbunja (Rubus Coreanus Miquel) is a wild berry to Rosaceae genus of which application areas have been expanded due to many health effects. Bokbunja contains carbohydrates, protein, fats and dietary fibers as major components and various flavonoids such as tannins, volatile components including organic acids, alcohols and hydrocarbons as minor ones. Those active compounds attribute to increase in immune function, antioxidant and anti-inflammatory activity. Recently pure compounds have been isolated from the extracts for the development of medicines. We report here on two cases of acne vulgaris that was successfully treated with Rubus Coreanus Miquel extracts.

Protective actions of Rubus coreanus ethanol extract on collagenous extracellular matrix in ultraviolet-B irradiation-induced human dermal fibroblasts

Solar ultraviolet (UV) irradiation leads to distinct changes in the skin connective tissues by degradation of collagen, which is a major structural component in the extracellular matrix. UV irradiation induces the production of matrix metalloproteinases (MMP) capable of attacking native fibrillar collagen and responsible for inhibiting the construction of collagenous extracellular matrix. In this study, we attempted to investigate the protective actions of Rubus coreanus ethanol extract (RCE) on the MMP production and the consequent procollagen/collagen degradation in UV-B-irradiated human dermal fibroblasts. The analytical data showed that Rubus coreanus ethanol extract was mostly comprised of cyanidin 3-rutinoside. Pre-treatment of fibroblasts with this extract inhibited UV-B-induced production of MMP-1, MMP-8 and MMP-13 in dose-dependent manners. In addition, Western blot analysis and immunocytochemical staining assay revealed that RCE markedly augmented the cellular levels of procollagen/collagen declined in UV-B-exposed dermal fibroblasts. These results demonstrate that RCE blocks UV-B-induced increase of the collagen degradation by inhibiting MMP production. Thus, RCE may act as an agent inhibiting excessive dermal collagen degradation leading to the skin photoaging.

Inhibitory Effect of Rubus coreanus on Compound 48/80-induced Anaphylactic Shock and Vascular Permeability in Vivo / 체질인류학회지

As an attempt to search for antiallergic and antiinflammatory activities, we examined he potency of Rubus coreanus water extract (RCWE). The effect of RCWE on systemic anaphylactic shock and vascular permeability was investigated. Results are 1) compound 48/40-induced mortality rate in mice were significantly inhibited by pretreatment of RCWE and compound 48/40-induced mean death time in mice were delayed by pretreatment of RCWE, 2) compound 48/40-induced mesenteric mast cell degranulation in mice were significantly inhibited by pretreatment of RCWE, 3) the increase of vascular permeability induced by compound 48/80 or anti-DNP IgE in rat was significantly inhibited by pretreatment of RCWE. These results suggest that RCWE contains substances with an activity to inhibit the systemic anaphylaxis and vascular permeability.

Inhibitory Effect of Rubus Coreanus on Compound 48/80- or Anti-DNP IgE-Induced Mast Cell Activation

BACKGROUND: The fruit of Rubus coreanus (RC), a perennial herb, has been cultivated for a long time as a popular vegetable. The anti-allergy mechanism of RC is unknown. The purpose of this study is to investigate the inhibitory effect of RC on compound 48/80- or anti-DNP IgE-induced mast cell activation. METHODS: For this, influences of RC on the compound 48/80-induced degranulation, histamine release, calcium influx and the change of the intracellular cAMP (cyclic adenosine-3',5' monophosphate) levels of rat peritoneal mast cells (RPMC) and on the anti-DNP IgE-induced histamine release of RPMC were observed. RESULTS: The pretreatment of RC inhibited compound 48/ 80-induced degranulation, histamine release and intracelluar calcium uptake of RPMC. The anti-DNP IgE-induced histamine release of RPMC was significantly inhibited by pretreatment of RC. The RC increased the level of intracellular cAMP of RPMC, and the pretreatment of RC inhibited compound 48/80-induced decrement of intracellular cAMP of RPMC. CONCLUSION: These results suggest that RC contains some substances with an activity to inhibit the compound 48/80- or anti-DNP IgE-induced mast cell activitation. The inhibitory effects of RC are likely due to the stabilization of mast cells by blocking the calcium uptake and enhancing the level of intracellular cAMP.