Search

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-β-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.

Phlorotannins have received much attention due to their ecophysiological importance and potential applications in the biotechnology and food industries. Antioxidant activity studies in seaweeds have mainly focused on in vitro assays; however, there is a paucity of data regarding the effect of brown algal phlorotannins on living cultured cells. The aim of the present study was to investigate both direct and protective effects of phlorotannin-rich extracts on cell viability and the cellular oxidative status of cultured liver cells HepG2 against oxidative stress induced by tert-butyl hydroperoxide (t-BOOH). Extracts of the Phaeophyceae Ascophyllum nodosum (Fucaceae) and Himanthalia elongata (Himanthaliaceae) were submitted to gastrointestinal digestion prior to incubation for 20 h in a HepG2 culture at physiological concentrations (0.5-50 οg mL-1). Various markers of cellular oxidative stress were then assessed, such as the generation of reactive oxygen species (ROS), antioxidant defences (concentration of reduced glutathione and activities of glutathione peroxidase, reductase and glutathione-S-transferase) and the levels of malondialdehyde as a marker for lipid peroxidation. The direct effect on cellular markers was assessed immediately after the incubation period, whereas for the protective effect, the incubation period was followed by a 3-h treatment with t-BOOH. The results indicated no effect on cell viability, and both extracts showed reduced levels of ROS and increased antioxidant defences in the direct treatment. Moreover, the extracts showed a significant protective effect against chemically induced oxidative stress in HepG2 cells by reducing ROS generation and enhancing antioxidant defences, hence supporting the utility of including brown algal extracts in functional food products. © 2012 Springer Science+Business Media B.V., This work was supported by grant AGL2007-64042/ALI and project CSD 2007-00063 from Programa Consolider-Ingenio from the Spanish Ministry of Science and Innovation (CICYT), an EU Erasmus grant under the Lifelong Learning Programme and a Mobility/Training grant from the Sea Change Networking Initiative of the Marine Institute, Ireland.

Phloroglucinol is an ubiquitous secondary metabolite encountered in a free state or polymerised as phlorotannins in brown macroalgae, and present in higher plants. FRAP and TEAC assays measured the antioxidant properties of phloroglucinol in non-biological conditions. Additionally, the biological effects of phloroglucinol (4-400 μM) were scrutinised using cellular oxidative stress markers, such as the generation of ROS, antioxidant defences (concentration of GSH and activities of GPx, GR and GST), and levels of MDA as a marker for lipid peroxidation. The direct effect was assessed immediately after an incubation period, whereas for the protective effect, the incubation period was followed by 3-h treatment with the pro-oxidant t-BOOH. The results indicated that despite having a higher radical scavenging capacity than Trolox after 30. min, phloroglucinol was not a suitable antioxidant standard for phlorotannins. Regarding the biological effects, phloroglucinol had no impact on cell viability, reduced levels of ROS and increased antioxidant defences in the direct treatment for most concentrations. The results of the protective effect were mitigated as phloroglucinol failed to protect from ROS generation but evoked a significant recovery of the stress-altered cellular antioxidant defences to restful conditions. Additionally, MDA levels were greatly reduced, preventing a radical chain oxidation., This work was supported by the Grant AGL2007-64042/ALI and Project CSD 2007-00063 from Programa Consolider-Ingenio from the Spanish Ministry of Science and Innovation (CICYT), an EU Erasmus Grant under the Lifelong Learning Programme and a Mobility/Training Grant from the Sea Change Networking Initiative of the Marine Institute, Ireland.

Interleukin-1 and interleukin-6 are principal cytokines involved in regulation of expression of acute-phase proteins. In the joint action of both cytokines IL-1 can suppress or enhance the IL-6-dependent induction of gene expression. Here, we report changes in the transcriptome profile of HepG2 cells exposed to IL-6 alone, or IL-1 and IL-6. Cytokine-responsive genes were identified by differential display analysis. Validation of observed changes in the transcript level was carried out using the slot blot method. Out of 88 cDNA species modulated by IL-6, only 38 represent different known genes whereas 18 clones match genomic clones in NCBI data with hypothetical cDNA sequences (the remaining 32 clones showed no homology with the database or represented several clones of the same gene). In the experiments with HepG2 cells prestimulated for 3 h with IL-1 and then stimulated with IL-6, 43 cDNA fragments were amplified. Twenty-three of them represent known genes while 10 clones have inserts matching hypothetical cDNA sequences in NCBI data. The identified transcripts modulated by IL-6 or both cytokines in HepG2 cells code for intracellular proteins of various function. The largest groups represent genes engaged in metabolism, protein synthesis and signaling pathways. Among all genes identified as differentially regulated under stimulation by IL-6, or IL-1/IL-6, six were detected in both types of stimulation. None of the typical genes coding for plasma acute phase proteins was identified in our experiments. This indicates that differential display cannot be used to characterize the profile of a given transcriptome. On the other hand, it is a useful technique for detection of new genes responding to IL-6 alone or IL-6 in combination with IL-1.