iridoids and pentaacetyl-geniposide

Gardenia, the fruit of Gardenia jasminoides Ellis, has been widely used to treat liver and gall bladder disorders in Chinese medicine. It has been shown recently that geniposide, the main ingredient of Gardenia Fructus, exhibits the anti-tumor effect. In this review, we discuss the anti-tumor effect and possible mechanisms of a derivative from Gardenia Fructus, penta-acetyl geniposide ((Ac)5GP). It has been demonstrated that (Ac)5GP plays more potent roles than geniposide in chemoprevention. (Ac)5GP decreased DNA damage and hepatocarcinogenesis induced by aflatoxin B1 (AFB1) by activating the phase II enzymes glutathione S-transferase (GST) and GSH peroxidase (GSH-Px). It reduced the growth and development of inoculated C6 glioma cells especially in pre-treated rats. In addition to the preventive effect, (Ac)5GP exerts its actions on apoptosis and growth arrest. Treatment of (Ac)5GP caused DNA fragmentation of glioma cells. (Ac)5GP induced sub- G1 peak through the activation of apoptotic cascades PKCdelta/JNK/Fas/caspase8 and caspase 3. Besides, p53/Bax signaling was suggested to be involved in (Ac)5GP-induced apoptosis, though its downstream cascades needs further clarified. (Ac)5GP has also been shown to inhibit DNA synthesis of tumor cells. It arrested cell cycle at G0/ G1 by inducing the expression of p21, thus suppressing the cyclin D1/cdk4 complex formation and the phosphorylation of E2F. The phosphorylation status of p53 on serine 392 correlated with the process of growth arrest. Evidences from the in vivo experiments showed that (Ac)5GP is not harmful to liver, heart and kidney. In conclusion, (Ac)5GP is highly suggested to be an anti-tumor agent for development in the future.

Topics: Animals; Antineoplastic Agents; Fruit; Gardenia; Glucosides; Humans; Iridoid Glucosides; Iridoids; Medicine, Chinese Traditional; Pyrans

Penta-acetyl geniposide [(Ac)(5)GP], an acetylated geniposide product from Gardenia fructus, has been known to have hepatoprotective properties and recent studies have revealed its anti-proliferative and apoptotic effect on C6 glioma cells. In this study, we first report the anti-metastastic effect of (Ac)(5)GP in the rat neuroblastoma line: C6 glioma cells. First (Ac)(5)GP exhibited an inhibitory effect on abilities of adhesion and motility by cell-matrix adhesion assay, wound healing assay and Boyden chamber assay. Second, the decreasing activity of matrix metalloproteinase-2 (MMP-2) was noted by gelatin zymography assay. Further analysis with semi-quantitative RT-PCR showed the mRNA levels of MMP-2 and membrane type I matrix metalloproteinase (MT1-MMP) were significantly reduced, while the tissue inhibitor of matrix metalloproteinase-2 (TIMP-2) was elevated by (Ac)(5)GP treatment. Further (Ac)(5)GP also exerted an inhibitory effect on phosphoinositide 3-kinase (PI3K) protein expression, phosphorylation of extracellular signal-regulated kinases 1 and 2 (ERK1/2) and inhibition of activation of transcription factor nuclear factor kappa B (NF-kappaB), c-Fos, c-Jun. These findings proved (Ac)(5)GP is highly likely to be a inhibiting cancer migration agent to be further developed in the future.

Topics: Animals; Base Sequence; Cell Line, Tumor; DNA Primers; Electrophoresis, Polyacrylamide Gel; Extracellular Signal-Regulated MAP Kinases; Glioma; Glucosides; Iridoid Glucosides; Iridoids; Matrix Metalloproteinase Inhibitors; Neoplasm Metastasis; Phosphatidylinositol 3-Kinases; Protease Inhibitors; Rats; Reverse Transcriptase Polymerase Chain Reaction; Signal Transduction

We have demonstrated the herbal derivative penta-acetyl geniposide ((Ac)(5)GP) induces C6 glioma cell apoptosis through the critical sphingomyelinase (SMase)/nerve growth factor (NGF)/p75 and its downstream signals. It has been reported mitogen-activated protein kinase (MAPK) mediates NGF synthesis induced by SMase activation. In this study, ERK, p38 and JNK are shown to mediate (Ac)(5)GP-induced glioma cell apoptosis and elevation of NGF and p75. Treatment of PD98059 (ERK-specific inhibitor), SB203580 (p38 MAPK inhibitor) and SP600125 (JNK inhibitor) decreases the elevation of NGF and p75 mRNA induced by (Ac)(5)GP, indicating possible transcription regulation via MAPKs. The results of nuclear extract blotting and EMSA further confirm (Ac)(5)GP maximally increases AP-1 and NF-kappaB DNA binding at 6h. Inhibition of ERK, p38 and JNK block the activation of AP-1 and NF-kappaB, suggesting these MAPKs are involved in (Ac)(5)GP-induced transcription regulation. We thereby used RT-PCR to analyze cells treated with (Ac)(5)GP, with or without AP-1 or NF-kappaB inhibitors. AP-1 inhibitor NDGA decreases NGF/p75 and expression of FasL and caspase 3 induced by (Ac)(5)GP, suggesting the importance of AP-1 in mediating NGF/p75 and their downstream apoptotic signals. However, FasL and caspase 3 do not change with the NF-kappaB inhibitor PDTC; NF-kappaB might be linked to other cellular events. Overall, we demonstrate that MAPK mediates (Ac)(5)GP-induced activation of AP-1, promoting the transcription of NGF/p75 and downstream apoptotic signals. These results further highlight the potential therapeutic effects of (Ac)(5)GP in chemoprevention or as an anti-tumor agent.

Topics: Animals; Anthracenes; Anti-Inflammatory Agents, Non-Steroidal; Antioxidants; Apoptosis; Cell Line, Tumor; Electrophoretic Mobility Shift Assay; Flavonoids; Gardenia; Glioma; Glucosides; Imidazoles; Immunoblotting; Iridoid Glucosides; Iridoids; Keratolytic Agents; Masoprocol; Mitogen-Activated Protein Kinases; Molecular Structure; Nerve Growth Factor; NF-kappa B; Peptide Fragments; Pyridines; Rats; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Transcription Factor AP-1; Transcriptional Activation

In our previous study, penta-acetyl geniposide ((AC)(5)GP) is suggested to induce tumor cell apoptosis through the specific activation of PKCdelta. However, the downstream signal pathway of PKCdelta has not yet been investigated. It was shown that JNK may play an important role in the regulation of apoptosis and could be a possible downstream signal of PKCdelta isoforms. In the present study, we investigate whether JNK is involved in (AC)(5)GP induced apoptosis. The result reveals that (AC)(5)GP induces JNK activation and c-Jun phosphorylation thus stimulating the expression of Fas-L and Fas. Using SP600125 to block JNK activation shows that (AC)(5)GP-mediated apoptosis and related proteins expression are attenuated. Furthermore, we find that the (AC)(5)GP induces apoptosis through the activation of JNK/Jun/Fas L/Fas/caspase 8/caspase 3, a mitochondria-independent pathway. The JNK pathway is suggested to be the downstream signal of PKCdelta, since rottlerin impedes (AC)(5)GP-induced JNK activation. Therefore, (AC)(5)GP mediates cell death via activation of PKCdelta/JNK/FasL cascade signaling.

Topics: Animals; Apoptosis; Caspase 3; Caspase 8; Caspase Inhibitors; Caspases; Cell Line, Tumor; Drug Screening Assays, Antitumor; Drugs, Chinese Herbal; Fas Ligand Protein; Gardenia; Glioma; Glucosides; Iridoid Glucosides; Iridoids; JNK Mitogen-Activated Protein Kinases; Membrane Glycoproteins; Mitochondria; Phosphorylation; Protein Kinase C; Pyrans; Rats; Signal Transduction; Taiwan; Up-Regulation

Penta-acetyl geniposide, (Ac)5-GP, the acetylated compound of geniposide, is able to inhibit the growth of rat C6 glioma cells in culture and in the bearing rats. Our recent data indicated that the induction of cell apoptosis and cell cycle arrest at G0/gap phase 1 (G1) by (Ac)5-GP might be associated with the induction of p53 and c-Myc, and mediated via the apoptosis-related bcl-2 family proteins. In this report, we further investigated the mechanism involved in the cell cycle arrest induced by (Ac)5-GP in C6 glioma cells. The inhibitory effect of (Ac)5-GP on the cell cycle progression of C6 glioma cells which arrested cells at the G0/G1 phase was associated with a marked decrease in the protein expression of cyclin D1, and an induction in the content of cyclin-dependent kinase (cdk) inhibitor p21 protein. This effect was correlated with the elevation in p53 levels. Further immunoprecipitation studies found that, in response to the treatment, the formation of cyclin D1/cdk 4 complex declined, preventing the phosphorylation of retinoblastoma (Rb) and the subsequent dissociation of Rb/E2F complex. These results illustrated that the apoptotic effect of (Ac)5-GP, arresting cells at the G0/G1 phase, was exerted by inducing the expression of p21 that, in turn, repressed the activity of cyclin D1/cdk 4 and the phosphorylation of Rb.

Topics: Animals; Antineoplastic Agents; Brain Neoplasms; Cell Cycle; Cell Line, Tumor; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p21; Cyclin-Dependent Kinases; Cyclins; Drugs, Chinese Herbal; Glioma; Glucosides; Iridoid Glucosides; Iridoids; Phosphorylation; Plant Extracts; Pyrans; Rats; Retinoblastoma Protein; Tumor Suppressor Protein p53

The suppressive effects of penta-acetyl geniposide, (Ac)(5)-GP, on the hepatotoxic lesions-induced by aflatoxin B(1) (AFB(1)) were investigated in male Wistar rats. Rats were divided into six groups: groups I and II served as normal and solvent control, respectively; group III was given AFB(1) (2 mg/kg body weight) alone; group IV was given (Ac)(5)-GP (2 mg/kg) alone; and groups V and VI received both AFB(1) (2 mg/kg body weight) and (Ac)(5)-GP (1 mg and 2 mg/kg body weight, respectively). Rats received treatments for 8 weeks, then were maintained on basal diet for 32 weeks. At the end of the experiment (week 40), the liver lesions (e.g. fatty change, eosinophilic and bile duct dilation) and preneoplastic changes in rats of groups V and VI were reduced when they were compared with group III. There were no liver lesions and preneoplastic changes in rats treated with (Ac)(5)-GP alone. Although no differences in the total number of gamma-glutamyl transpeptidase (GGT)-positive foci was observed between the groups treated with AFB(1) along with or without (Ac)(5)-GP, the treatment of (Ac)(5)-GP significantly reduced the number of AFB(1)-induced GGT positive foci (with diameter larger than 0.3 mm). These results indicated that the protective effect of (Ac)(5)-GP on early hepatocarcinogenesis-induced by AFB(1) was associated with the inhibition of GGT foci development.

Topics: Aflatoxin B1; Animals; Anticarcinogenic Agents; Drugs, Chinese Herbal; gamma-Glutamyltransferase; Glucosides; Iridoid Glucosides; Iridoids; Liver; Liver Neoplasms, Experimental; Male; Precancerous Conditions; Pyrans; Rats; Rats, Wistar

Experimental data are provided which demonstrate the inhibitory effects of penta-acetyl geniposide, (Ac)5-GP, on the growth and development of C-6 glioma cells inoculated into rats. In the pretreatment experiments, (Ac)5-GP prolonged the latency period of T50 (time for 50% tumour incidence). At week 7, the growth inhibition was 41% with 5 mg/kg and 75% with 10 mg/kg of (Ac)5-GP. In the post-treatment experiments, growth inhibition was less. No significant hepatotoxic effects were observed in the treated group, indicated by the constant levels of serum enzymes (e.g. asparate aminotransferase). We suggest that (Ac)5-GP is a potent chemopreventive agent on glioma cells.

Topics: Animals; Cell Division; Cells, Cultured; Drugs, Chinese Herbal; Glioma; Glucosides; Iridoid Glucosides; Iridoids; Liver; Male; Pyrans; Rats; Rats, Wistar

A new compound, penta-acetyl geniposide ((Ac)5-GP), was obtained from modified extract of Gardenia fructus (San-Jee-Chee in Chinese). The structure of the compound was identified as 1-(beta-D-2',3',4',6'-tetraacetyl-glucopyrannosyloxyl)-1,4a, 5,7a-tetrahydro-7-(acetomethyl)-cyclopentapyran-4-carboxylic acid methyl ester, according to the spectral data. The inhibitory effects of (Ac)5-GP on aflatoxin B1 (AFB1)-induced cytotoxicity and DNA damage were studied. In the investigation of the inhibitory effect of (Ac)5-GP on AFB1-cytotoxicity, the plating efficiency of C3H10T1/2 cells in S-9 activation system was increased. In addition, (Ac)5-GP inhibited the DNA damage of AFB1-treated C3H10T1/2 cells, and it interfered with the inhibitory effect of DNA synthesis caused by AFB1. These results suggest that the reduced DNA damage and the increased DNA synthesis from cultured C3H10T1/2 cells are important mechanisms for the inhibition of AFB1-cytotoxicity by (Ac)5-GP.

Topics: Aflatoxin B1; Animals; Antimutagenic Agents; Cell Line; DNA Damage; DNA Replication; Drugs, Chinese Herbal; Glucosides; Iridoid Glucosides; Iridoids; Mice; Mice, Inbred C3H; Plants, Medicinal; Pyrans