Monascus pilosus-fermented black soybean inhibits lipid accumulation in adipocytes and in high-fat diet-induced obese mice

Objective:To explore the anti-obesity effects and the mechanism of action of Monascus pilosus(M.pilosus)-fermented black soybean(MFBS)extracts(MFBSE)and MFBS powders(MFBSP)in adipocytes and high-fat diet(HFD)-induced obese mice,respectively.Methods:Black soybean was fermented with M.pilosus,and the main constituents in MFBS were analyzed by HPLC analysis.In vitro,MFBSE were examined for anti-adipogenic effects using Oil-Red O staining.In vivo,mice were fed a normal-fat diet(NFD)control,HFD control or HFD containing 1 g/kg MFBSP for 12 weeks,and then body weight gain and tissues weight measured.Real-time PCR and western blot assay were used to determine the mechanism of anti-adipogenic effects.Results:MFBSE inhibited lipid accumulation in 3T3-L1 adipocytes without exerting cell cytotoxicity.MFBSP treatment in HFD-fed mice significantly decreased the body weight gain compared with the HFD control mice.MFBSE and MFBSP treatment resulted in significantly lower mRNA levels of adipogenesis-related genes,such as peroxisome proliferator-activated receptorγ(PPARγ),fatty acid-binding protein 4(FABP4),and fatty acid synthase(FAS),in adipocytes and in white adipose tissue(WAT)of HFD-induced obese mice.Conclusions:These results suggest that the anti-obesity effects of MFBS are elicited by regulating the expression of adipogenesis-related genes in adipocytes and WAT of HFDinduced obese mice.

Green coffee bean extract improves obesity by decreasing body fat in high-fat diet-induced obese mice

Objective:To evaluate possible lipid catabolism and body fat regulation effects of 3-caffeoylquinic acid in Green coffee bean extract(GCBE) in high-fat diet(HFD)-induced obese mice.Methods:Obesity was induced in mice using a HFD for four weeks.Then,mice were fed only HFD or HFD with GCBE at 50,100,and 200 mg/kg.Fatty acid synthesis mechanism regulation of body fat was investigated through real-time PCR and Western blot assay.Body fat reduction was measured through dual-energy X-ray absorptiometry.Results:In HFD-induced obese mice,GCBE treatment significantly decreased body weight gain,liver weight and white adipose tissue weights with regulation of adipose tissue lipolysis hormones,like adiponectin and leptin.GCBE treatment decreased mR NA expression levels of adipogenesis and adipocyte metabolism related genes in adipose tissues and the liver,and decreased the corresponding protein expression.Dual energy X-ray absorptiometry measurements were used to compare body fat between mice on high-fat and those treated with GCBE.GCBE treated mice had a lower fat mass compared to HFD alone fed mice and relative body weight and fat mass were markedly decreased.Conclusions:GCBE has a potential anti-obesity effect with lowering body fat accumulation by regulating adipogenesis and lipid metabolism-related genes and proteins in WAT and liver.

Protective effects of a polymethoxy flavonoids-rich Citrus aurantium peel extract on liver fibrosis induced by bile duct ligation in mice

Objective: To evaluate the possible protective effect of Citrus aurantium peel extract(CAE) against apoptosis in cholestatic liver fibrosis induced by bile duct ligation in mice. Methods: Male ICR mice were divided to 5 groups: 1) Control group(Sham-operated mice), 2) Cholestatic liver injury group induced by bile duct ligation(BDL), 3) BDL mice treated with silymarin(200 mg/kg) for 4 weeks, 4) BDL mice treated with 50 mg/kg CAE for 4 weeks, 5) BDL mice treated with 200 mg/kg CAE for 4 weeks. Mice were sacrificed and liver fibrosis was evaluated by serum and hepatic tissue biochemistry tests and liver histopathological examination. Effects of CAE on inflammation and apoptosis gene regulation were investigated through real-time PCR. CAE effect on lipid metabolism related signaling was determined by western blot analysis. Results: In BDL mice, administration of CAE for 4 weeks markedly attenuated liver fibrosis based on histopathological alteration. Serum and hepatic tissue biochemistry results revealed that CAE(50 and 200 mg/kg) decreased the levels of alanine transaminase, aspartate transaminase, gamma glutamyl transferase, total bilirubin, nitric oxide, and thiobarbituric acid reactive substances. Real-time PCR and western blot analysis showed that CAE regulated inflammation, apoptosis, and lipid metabolism factors increased by BDL. Interleukin family, tumor necrosis factor α, and related apoptosis factors m RNA levels were increased by BDL treatment. However, these increases were suppressed by CAE administration. In addition, CAE effectively increased phosphorylation of AMPactivated protein kinase, nuclear factor E2-related factor 2, and related cytoprotective proteins. Conclusions: CAE can efficiently regulate BDL-induced liver injury with antioxidant, antiinflammatory, and antiapoptotic activities.

Antioxidant activity and free radical scavenging activities of Streptomyces sp.strain MJM 10778

Objective:To investigate the antioxidant activity of soil-borne aetinobacteria.Methods:The total phenolic contents,the level of antioxidant potential by DPPH radical scavenging activity,MO scavenging activity,and ABTS radical scavenging activity in ethyl acelale extract were determined.Results:The 16 S rDNA sequencing analysis revealed that Streptomyces sp.strain MJM 10778.which was isolated from Hambak Mountain.Korea,has 99.9% similarity to Streptomyces misionensis(S.misionenis) NBRC 13063.The physiological and the morphological test revealed that the strain MJM 10778 has different characteristics from the strain NBRC.13063.The entire antioxidant assay with the ethyl acelale extract displayed good radical scavenging activity.The IC_(50) values of the strain MJM 10778 extract on DPPH,.NO.and ABTS radicals were identified to he 92.8 μg/mL,0.02 μg/ml,and 134.9 μg/mL,respectively.The ethyl acetate extract of the strain MJM 10778 showed an 81.500% of cell viability at 100 μg/mL in Raw264.7cell viability assay.Conclusions:The results obtained suggesl that the ethyl acetate extract of Streptomyces sp.strain MJM 10778 could be considered as a potential source of drug for the diseases that is caused by free radicals with its anti-oxidant activities and low cytotoxicity.

Roots extracts of Adenophora triphylla var.japonica improve obesity in 3T3-L1 adipocytes and high-fat diet-induced obese mice

Objective:To investigate the anti-obesity activity and the action mechanism of the roots of Adenophora triphylla var.japonica extract(ATE) in high-fat diet(HFD)-induced obese mice and 3T3-LI adipocytes.Methods:The roots of Ademption triphylla were extracted with 70%ethanol.To demonstrate the compounds,linoleic acid was analyzed by using gas chromatography:and the anti-obesity effects and possible mechanisms of ATE were examined in 3T3-L1 adipocytes and HFD-induced obese mice.Results:Treatment with ATE inhibited the lipid accumulation without cytotoxicity in 3T3-L1 adipocytes.Furthermore,200 and 400 mg/kg ATE treatment significantly decreased the body weight gain,white adipose tissues(WATs) weight and plasma triglyceride level,while 100 and 200 mg/kg ATE treatment increased the plasma high-density lipoprotein cholesterol level in the HFD-induced obese mice,as compared with the HFD group.Treatment with 200 and 400 mg/kg ATE also lowered the size of adipocytes in adipose tissue and reduced the lipid accumulation in liver.ATE treatment showed significantly lower expression level of adipogenesis-relatcd proteins,such as peroxisome proliferator-activated receptor y,fatty acid binding protein(aP2),fatty acid synthase in 3T3-L1 adipocytes;and furthermore,decreased peroxisome proliferatoractivated receptor γ,aP2,fatty acid synthase,sterol regulatory element binding protein-1c,and lipoprotein lipase mRNA expression levels in WAT of the HFD-induced obese mice.Conclusions:These results suggested that the ATE has an anti-obesity effect,which may be elicited by regulating the expression of adipogenesis and lipogenesis-related genes and proteins in adipocytes and WAT of the HFD-induced obese mice.

An easy PCR-based genome-walking method for getting the unknown 5’flanking region of a Scenedesmus sp.

Objective:To develop the current single primer PCR-based genome-walking method with Scenedesmus sp.Methods:The unknown 5’and/or 3’flanking regions for a specific conserved sequence were optimized and the current single primer PCR-based genome-walking method were developed.Alignment was between the related species of microalga and Scenedesmus sp.For 18S rDNA,we selected the species Scenedesmus sp.,Chlorella sp.,and Chlamydomonas sp.For the rbcL gene from the chloroplast genome,alignment was done between Scenedesmus sp.,and Chlamydomonas sp.Results:Obtaining a small conserved sequence for any gene family is something that can be achieved quite easily.However,identifying the whole gene is often difficult.After investigating and testing,some of the current protocols using to get the unknown 5’and/or 3’flanking regions for a specific conserved sequence,we developed the current single primer PCR-based genome-walking method.We performed two consecutive PCR reactions;band extraction and the PCR product were sequenced.We got our results by testing the method on three genes from the total DNA of Scenedesmus sp.;two genes had a fully known sequence in gene bank(18S rDNA and rbcL),but the third one has not yet been identified(rbcS).We designed our primers based on the alignment between the related species and to each other.We also tested two different DNA polymerases Ex Taq and TLA polymerase.Conclusions:Results from our study suggest that Ex Taq is the most suitable polymerase for the current protocol.

Effect of fermentation with Monascus pilosus on the antioxidant activities and phenolic acid contents of adzuki bean(Vigna angularis)

Objective:To enhance physiological activities of adzuki bean(Vigna angularis)via fermentation with Monascus pilosus(M.pilosus).Methods:The adzuki bean fermentation conditions with M.pilosus were optimized,and the effect of Monascus-fermentation on the antioxidant capacity and phenolic acid contents of adzuki bean was investigated.Results:Optimal fermentation conditions were determined by the production of monacolin K.The highest monacolin K production was observed in 5%inoculum sized on day 15 in fermentation.Free and bound phenolic acids were isolated from native and fermented adzuki bean.A 1.9-fold decrease was observed in bound p-coumaric acid content,whereas the contents of bound ferulic and sinapic acids were increased by 28-and 1.7-fold,respectively.However,the contents of free phenolic acids such as p-coumaric,ferulic,and sinapic acids were increased by 2.6-,5.2-,and 7.2-fold,respectively.The fermentation of adzuki bean by M.pilosus enhanced the activities of DPPH■radical scavenging,ferrous ion-chelating,nitric oxide scavenging,and ferric antioxidant reducing activities 2.2-,1.7-,1.2-,and 1.8-fold,respectively.Conclusions:Results from our study suggest that the contents of p-coumaric,ferulic,and sinapic acids in adzuki bean were highly increased by fermentation with M.pilosus,resulting in enhanced various antioxidant activities.