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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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Background: Orostachys japonicus has traditionally been used for antifever, anti-inflammation, and anticancer. O. japonicus has not yet been studied for cosmetic effects. Objectives: In this study, O. japonicus extracts were investigated to verify the possibility of them as anti-aging cosmetic agents. Materials and Methods: O. japonicus were used to prepare experimental extracts using 90% methanol (MeOH) or ethanol (EtOH) as solvent. Safety and efficacy tests applied in this work are as follows: Folin-Denis' method for total phenolic contents, liquid chromatography for component analysis, 3-[4,5-dimethyl thiazol-2-yl]-5-(3-carboxymethoxyphenyl) -2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay for cytotoxicity, 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), and ferric reducing antioxidant power (FRAP) assay for antioxidant activity, biochemical inhibition assays of elastase activity, collagenase activity, matrix metalloproteinase-1 (MMP-1) and MMP-3 mRNA expression), and MMP-1 protein production for anti-wrinkle activity, and tyrosinase activity inhibition assay for whitening activity. Results: This research revealed that O. japonicus extracts contained various flavonoids, showed no adverse cytotoxicity up to 300 or 500 μg/mL in HS68 and B16F10, exhibited significant antioxidant activity, exerted remarkable anti-wrinkle activity, and represented useful whitening activity. Conclusion: This study suggested that O. japonicus extracts could be utilized to develop anti-aging cosmetic agents exerting useful whitening activity as well as excellent antioxidant and anti-wrinkle activities. [ABSTRACT FROM AUTHOR]

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-a, and interleukin-6 production was observed. Conclusions: O. japonicus inhibits oxidative and inflammatory reactions with potentially positive health-related effects.

Objective: 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-hexane, 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 anti-biofilm formation of MRSA and antibacterial agents in the future.

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‐1β, 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. [ABSTRACT FROM AUTHOR]

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-G1 and S phase. Among the upstream and downstream proteins involved in anti-cancer activity, the level of B cell lymphoma-2 decreased, and the bcl-2-associated x protein increased. The level of pro-caspase-3, pro-caspase-8, and pro-caspase-9 decreased, while the level of cleaved-caspase-3, cleaved-caspase-8, and cleaved-caspase-9 increased. Moreover, the phosphorylation, that is, activation of extracellular signal regulated kinase 1/2, Jun-N-terminal kinase, and p38 increased. Conclusions: 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.

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. [ABSTRACT FROM AUTHOR]

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. Orostachys japonicus (Crassulaceae), a perennial herb as a medicinal plant, has been used as a treatment of folk medication. In this study, we investigated the effect of OJE (ethyl acetate fraction from O. japonicus) on cell proliferation as well as its apoptotic pathway and cell cycle progression in the OVCAR‐3 human ovarian cancer cell line. [ABSTRACT FROM AUTHOR]

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-G1 and S phase. Among the upstream and downstream proteins involved in anti-cancer activity, the level of B cell lymphoma-2 decreased, and the bcl-2-associated x protein increased. The level of pro-caspase-3, pro-caspase-8, and pro-caspase-9 decreased, while the level of cleaved-caspase-3, cleaved-caspase-8, and cleaved-caspase-9 increased. Moreover, the phosphorylation, that is, activation of extracellular signal regulated kinase 1/2, Jun-N-terminal kinase, and p38 increased. Conclusions: 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. [ABSTRACT FROM AUTHOR]

We investigated the anti-inflammatory effect of the hexane fraction from Orostachys japonicus (OJH) in LPS (lipopolysaccharide)-stimulated RAW 264.7 cells. Pretreatment with OJH dose-dependently reduced the cellular NO (nitric oxide) concentration and also inhibited expression of iNOS (inducible nitric oxide synthase) protein and mRNA. By the prevention of IκBα (inhibitory factor kappa B alpha) phosphorylation and degradation, OJH inhibited LPS-induced NF-κB (nuclear factor-kappa B) activation. OJH had no effect on the LPS-induced phosphorylation of ERK (extracellular signal-regulated kinase), JNK (c-Jun N-terminal kinase), or p38, whereas it attenuated the phosphorylation of Akt in a dose-dependent manner. In addition, OJH suppressed the LPS-induced expression of LPS receptors CD14 and TLR4 (toll like receptor 4). These results suggest that OJH may interrupt LPS-induced pro-inflammatory cascades through inhibition of NF-κB and Akt activation.

The herb Orostachys japonicus has been traditionally used to treat chronic diseases, such as hepatitis, hemorrhoids, and cancer, in Asia. In this study, we investigated the effect of Orostachys japonicus water extract (OJWE) on the receptor activator of nuclear factor-κB ligand (RANKL)-induced osteoclast differentiation and bone loss. We found that OJWE inhibited RANKL-induced osteoclast differentiation in a dose-dependent manner without affecting bone resorption in bone marrow-derived macrophage cells. Interestingly, OJWE significantly reduced serum levels of C-terminal telopeptide of type 1 collagen and tartrate-resistant acid phosphatase (TRAP) 5b, markers of bone resorption and osteoclast number, respectively, in an animal model of bone loss. Furthermore, OJWE suppressed the RANKL-induced up-regulation of nuclear factor of activated T cells cytoplasmic 1 (NFATc1) expression, and activation of the p38 signaling pathway, but prevented the RANKL-mediated down-regulation of interferon regulatory factor-8 (IRF-8), which is known to be an anti-osteoclastogenic factor that represses NFATc1 expression. We also identified gallic acid and quercetin-3-O-β-D-glucoside as the OJWE components that inhibit RANKL-induced osteoclast differentiation. These results suggest that OJWE inhibits osteoclast differentiation by inhibiting RANKL-induced NFATc1 expression, which prevents osteoclast differentiation and bone loss. The present study elucidated a mechanism of action underlying the inhibitory effect of OJWE on osteoclast differentiation. Our findings suggest that O. japonicus has therapeutic potential for use in the treatment of bone diseases. [ABSTRACT FROM AUTHOR]

Orostachys japonicus has been used in traditional medicine as an anticancer agent. The present study aimed to investigate the mechanism by which O. japonicus extract affects the expression of matrix metalloproteinase (MMP)-2 and MMP-9, its association with the expression of the induc-ible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) genes in phorbol myristate acetate-differentiated THP-1 human monocytic leukemia cells and how it mediates the nuclear factor (NF)-κB and mitogen-activated protein kinase (MAPK) pathways. Cell proliferation was analyzed by MTT assay, mRNA expression was detected by quantitative polymerase chain reaction and protein expression was measured by western blot analysis. It was demonstrated that O. japonicus suppressed the mRNA expression of MMP-2 and MMP-9. In addition, O. japonicus was found to downregulate iNOS and COX-2 transcription and translocation. Furthermore, O. japonicus inhibited NF-κ;B p65 activity as well as the phosphorylation of p38 MAPK, MAPK kinase (MEK) and extracellular signal regulated kinase (ERK). The present results suggested that O. japonicus inhibited not only MMP-2 and MMP-9 mRNA expression, but also iNOS and COX-2 gene expression, suppressed NF-κB activation and reduced phosphorylation of p38 MAPK, MEK and ERK. The present results therefore indicated that O. japonicus was able to inhibit the expression of MMP-2 and MMP-9 and suppress the transcription and translocation of iNOS and COX-2 by directly inhibiting the activation of NF-κB and the phosphorylation of the MAPK pathway in THP-1 cells. [ABSTRACT FROM AUTHOR]

Tight junctions (TJs) are a mode of cell-to-cell adhesion in epithelial or endothelial cells, and serve as a physical barrier to maintenance of homeostasis in body by controlling paracellular transport. Claudins are the most important molecules of the TJs, but paradoxically these proteins are frequently over-expressed in cancers and their overexpression is implicated in the invasive potential of cancer. Hence, we investigated the effects of flavonoids extracted from Orostachys japonicus A. Berger (FEOJ) on TJs and the expression of claudins as well as cancer invasion along with in LnCaP human prostate cancer. FEOJ suppressed cancer cell motility and invasiveness at the concentrations where FEOJ did not show anti-proliferative activity. FEOJ increased transepithelial electrical resistance (TER) associated with tightening TJs, and suppressed expression of claudin proteins. Furthermore, FEOJ suppressed the activities of MMP-2 and -9 in a dose-dependent manner, which came from the activation of tissue inhibitor of metalloproteinases (TIMPs) by FEOJ. FEOJ suppressed migration and invasion by suppressing PI3K/Akt signaling pathway. Taken together, this study suggest that FEOJ suppresses cancer migration and invasion by tightening TJs through the suppression of claudin expression, and by suppressing MMPs in LnCaP human prostate cancer cells, which at least in part results from the suppression of PI3K/Akt signaling pathway. [ABSTRACT FROM AUTHOR]

Highlights: [•] OJH inhibits LPS-induced pro-inflammatory molecules. [•] OJH inhibits LPS-induced activation of NF-κB and PI3K signaling. [•] OJH is an interesting target for the treatment of inflammatory disease. [Copyright &y& Elsevier]

Abstract: Inhibition of fatty acid synthase (FAS) has been proposed as a new therapeutic target for the treatment of cancer. In our preliminary study, a MeOH extract of the aerial parts of Orostachys japonicus A. Berger (Crassulaceae) inhibited FAS activity (>95% inhibition at 62.5μg/mL). Through bioassay-guided fractionation, we isolated three cerebrosides (1–3), including one new compound (1), and seven glyceroglycolipids (4–10) from a FAS inhibitory hexane-soluble fraction of the MeOH extract of O. japonicus. On the basis of physicochemical and spectroscopic analyses including 1D and 2D NMR, HRFABMS, and ESIMS/MS, the structure of the new compound 1 was 1-O-(β-d-glucopyranosyl)-(2S,3S,4R,6Z,10E)-2-[(2′R,6′Z,9′Z)-2-hydroxyeicosa-6′,9′-dienoylamino)]-1,3,4-trihydroxytetracosa-6,10-diene, and it was named orostachyscerebroside A. Compounds 1–10 were tested for their in vitro FAS inhibitory activity, and compounds 1, 4, 5, 6, 7, and 8 inhibited the enzyme with micro-molar IC50 values both in the incorporation of [3H] acetyl CoA (from 52.2 to 108.0μM) and in the oxidation of NADPH (from 19.5 to 30.3μM). Interestingly, compounds 1, 4, 5, 6, 7, and 8 with FAS inhibitory activity showed cytotoxic activity against HL-60 (human leukemia) and A549 (human lung adenocarcinoma) cancer cells, while none of them displayed cytotoxicity against MCF-7 (human breast adenocarcinoma) cells. This is the first report on the inhibition of FAS by glycolipids from O. japonicus and their cytotoxic activity and may provide a scientific basis for the folk remedy using the plant to treat cancer. [Copyright &y& Elsevier]

Abstract: Ethnopharmacological relevance: Orostachys japonicus A. Berger (O. japonicus) is known to reduce the risk of many diseases. Aim of the study: We investigated the anti-inflammatory effects of the dichloromethane (DCM) fraction from O. japonicus (OJD) in LPS-stimulated RAW 264.7 cells. Materials and methods: NO was measured using the Griess method. Key pro-inflammatory cytokines and mediators including IL-1β, TLR4, iNOS, and COX-2; 2 important pro-inflammatory transcription factors, NF-κB p65 and IκBα; and MAPKs such as ERK1/2, JNK, and p38 were analyzed by Western blotting. Results: OJD significantly inhibited NO production, IL-1β, TLR4, iNOS, and COX-2 expression in LPS-stimulated cells. Additionally, it inhibited LPS-induced NF-κB p65 activation via inhibition of IκBα phosphorylation. Furthermore, phosphorylation of p38 and JNK was suppressed by OJD in a dose-dependent manner in the LPS-stimulated RAW 264.7 cells. Conclusions: Our data suggest that OJD inhibits the inflammatory response via suppression of NF-κB activation and MAPK signaling. [Copyright &y& Elsevier]

Abstract: Orostachys japonicus cultivated in the Republic of Korea was analysed for flavonoid content via HPLC coupled to MS/MS. Amongst 16 compounds that were characterised, eight flavonoids and one alkaloid were characterised for the first time: two procyanidin dimer gallate isomers (1 and 2), epigallochatechin-3-gallate (3), two procyanidin dimer digallate isomers (4 and 9), quercetin 3-O-rhamnosyl-7-O-glucoside (6), myricetin 3-O-glucoside (10), kaempferol (16) and N 1,N 5,N 10-tri-p-(E,E,E)-coumaroylspermidine (15). The identified compounds were quantified by HPLC–UV/DAD. The antioxidant activity of the O. japonicus flavonoids was determined via 2,2-diphenyl-1-picrylhydrazyl radical (DPPH•), 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulphonic acid) radical cation (ABTS•+) and nitric oxide radical (NO•) scavenging assays. [ABSTRACT FROM AUTHOR]

The n-butanol ( n-BuOH) fraction of Orostachys japonicus A. Berger (Crassulaceae) significantly inhibited calpain activity. Through the activity-guided isolation from the n-BuOH fraction, herbacetin 8- O-α-D-ribopyranoside ( 1), kaempferol ( 2), quercetin ( 3), afzelin ( 4), astragalin ( 5), isoquercetin ( 6) and quercitrin ( 7) were obtained. Their structures were determined by spectroscopic techniques. Among them, compound 3 and 5 had significant calpain inhibitory activities. [ABSTRACT FROM AUTHOR]

Abstract: Calpains are calcium-dependent proteases that cleave a variety of intracellular substrates. The overactivation of μ-calpain is associated with a wide range of disease conditions. To search for calpain inhibitors from natural products, the phytochemical constituents of the ethyl acetate fraction of the whole plant of Orostachys japonicus (Crassulaceae) were studied. The various chromatographic separation of this fraction led to the isolation of a new tannin, (−)-epicatechin 5-gallate (1) along with 9 known compounds. Their structures were elucidated by spectroscopic and chemical analyses. Among them, (−)-epicatechin 5-gallate (1) and kaempferol (9) exhibited moderate inhibitory activity against μ-calpain with IC50 values of 18.0±2.9 and 15.4±2.0 µg/ml, respectively. [Copyright &y& Elsevier]

Abstract: The effects of methanol extract and gallic acid (3,4,5-trihydroxybenzoic acid) of Orostachys japonicus A. Berger on hepatic drug metabolizing enzymes and lipid peroxidation were investigated in rats treated with bromobenzene. The methanol extract of Orostachys japonicus reduced the activities of phase I enzymes, aminopyrine N-demethylase and aniline hydroxylase, that had been increased by i.p. injection of bromobenzene. Gallic acid isolated from Orostachys japonicus also reduced the aniline hydroxylase activity, while it did not affect the aminopyrine N-demethylase activity. The methanol extract and gallic acid restored the activity of epoxide hydrolase which had been decreased by bromobenzene. Hepatic glutathione content was lowered, along with increase in hepatic lipid peroxide, by bromobenzene administration. The hepatic lipid peroxidation induced by bromobenzene was prevented with the methanol extract and gallic acid of Orostachys japonicus. However, the decrease in glutathione was not altered by gallic acid. The present results suggest that the methanol extract and gallic acid of Orostachys japonicus may protect liver from bromobenzene toxicity through, at least in part, inhibiting the cytochrome P450-dependent monooxygenase activities and enhancing the activity of epoxide hydrolase. Antioxidant effect also may contribute to the protection of Orostachys japonicus against the bromobenzene-induced hepatotoxicity. [Copyright &y& Elsevier]

From the whole plants of Orostachys japonicus, kaempferol, quercetin, astragalin, quercitrin, isoquercitrin, cynaroside, afzelin, 3- O-α-L-rhamnosyl-7- O-β-D-glucosyl kaempferol, and 3,7-di- O-β-D-glucosyl kaempferol were isolated and characterized by spectral data. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]