ABSTRACT
In the present study, to identify the effective components of Chinese traditional herbs, Euphorbia hylonoma Hand.-Mazz. (Euphorbiaceae), a folk herb that has been used among the Qinling mountain area for hundreds of years, was investigated. 3,3'-Di-O-methyl ellagic acid-4'-O-ß-d-xylopyranoside (JNE2), an ellagic acid derivative, was isolated from the acetone extract of the herb and its antitumor activity against human hepatoma HepG2 cells was detected in vitro. The results showed that JNE2 inhibited the proliferation of HepG2 cells in a dose- and time-dependent manner and blocked the cell cycle at the G1/S phase. A high dosage of JNE2 induced apoptosis of the tumor cells, but no significant differences were identified between the treatment groups. The invasiveness of HepG2 cells was also inhibited by JNE2. The mechanism of the antitumor effect of JNE2 at the molecular level was presumed to be due to the upregulation of the protein expression of Bax and caspase-3, and the downregulation of the protein expression of Bcl-2 and CCND1. The results suggested that JNE2 is a potential antitumor agent that merits further investigation.
ABSTRACT
ETHNOPHARMACOLOGICAL RELEVANCE: Panax ginseng is a well-known traditional Chinese medicine and has been used for treatment of various diseases for more than four thousand years in Asia. Ginseng saponins or ginsenosides, the active constituents are reported to possess antidiabetic activity, but their antihyperglycemic mechanisms are not fully elucidated. In the present study, the mechanisms of action of ginsenoside Re were investigated in vitro models. MATERIALS AND METHODS: 3T3-L1 cells were chosen as the model to investigate the molecular mechanisms of action of ginsenoside Re. Influence of ginsenoside Re on the adipogenesis was examined by determining TG levels in 3T3-L1 adipocytes by the method of TG oxidation enzyme. Glucose uptake in 3T3-L1 cells stimulated by insulin in the absence or presence of ginsenoside Re were quantified by measuring (3)H-2-deoxy-d-glucose levels. Cytokine proteins released into the medium including adiponectin and TNF-α were tested using respective ELISA kits. In addition, real time RT-PCR was conducted to investigate the expression changes of PPAR-γ and its responsive genes, ap2, adiponectin, IRS-1, GLUT4 and TNF-α. And western blot analysis was performed to determine the translocation of GLUT4. Finally, effects of ginsenoside Re on NO production in 3T3-L1 adipocytes and in macrophages were investigated through measurement of nitrite concentration by Griess reagent. RESULTS: Ginsenoside Re induced adipogenesis of 3T3-L1 adipocytes by accumulating TG, increased glucose uptake and up-regulated PPAR-γ2, IRS-1, ap2 and adiponectin genes expressions. Meanwhile, Re also increased production and release of adiponectin. Although having no effects on GLUT4 gene expression, Re facilitated GLUT4 protein translocation to the membranes. In addition, Re inhibited the expression and release of TNF-α. Finally, Re did not show inhibitory effects on NO production both in 3T3-L1 cells stimulated by LPS, TNF-α and IFN-γ and in LPS-stimulated mouse peritoneal macrophages. CONCLUSIONS: Ginsenoside Re exhibited the action of reducing insulin resistance through activation of PPAR-γ pathway by directly increasing the expressions of PPAR-γ2 and its responsive genes, adiponectin, IRS-1, ap2, inhibiting TNF-α production and facilitating the translocation of GLUT4 to promote glucose uptake and disposal in 3T3-L1 adipocytes.
Subject(s)
Ginsenosides/pharmacology , Insulin Resistance , PPAR gamma/genetics , Tumor Necrosis Factor-alpha/antagonists & inhibitors , 3T3-L1 Cells , Adipogenesis/drug effects , Adiponectin/genetics , Adiponectin/metabolism , Animals , Cells, Cultured , Fatty Acid-Binding Proteins/genetics , Gene Expression Regulation/drug effects , Glucose/metabolism , Glucose Transporter Type 4/genetics , Insulin Receptor Substrate Proteins/genetics , Macrophages, Peritoneal/drug effects , Macrophages, Peritoneal/metabolism , Male , Mice , Nitric Oxide/metabolism , Tumor Necrosis Factor-alpha/geneticsABSTRACT
OBJECTIVE: To study the chemical constituents from flowers of Koelreuteria paniculata. METHODS: Column chromatography and spectral analysis were used to isolate and identify the constituents. RESULTS: The EtOAc fraction from flowers of Koelreuteria paniculata was separated and purified. Nine compounds were obtained and identified as:sitosterol glucoside (I), gallic acid (II), kaempferol (III), luteolin (IV), kaempferol-3-O-(6"-acetyl)-beta-D-glucopyranoside (V), hyperoside-2"-O-acetyl (VI), hyperoside-2"-O-galloyl (VII), hyperoside (VIII), kaempferol-3-O-D-glucopyranoside (IX). CONCLUSION: Nine compounds are isolated for the first time from flowers of Koelreuteria paniculata. Compounds IV, V, VI and IX are isolated from this genus for the first time.
