Chemical differentiation of two taste variants of Gynostemma pentaphyllum by using UPLC-Q-TOF-MS and HPLC-ELSD.

To differentiate the sweet and bitter taste variants of a Chinese medicinal tea Gynostemma pentaphyllum (GP), a method for the quantitative analysis of ginsenosides Rb(1), Rb(3), Rd, and F(2) in GP by using UPLC-Q-TOF-MS was developed. According to the different contents of the four ginsenosides, chemical differentiation of the two taste variants of GP was achieved by principal component analysis (PCA). A supplementary quantitative analysis method of using HPLC-ELSD for determination of 20(S)-panaxadiol in the hydrolysates of GP was also developed. Similarly, chemical differentiation based on different amounts of 20(S)-panaxadiol was established and the result was well consistent with that based on the analysis of the four ginsenosides. It was found that the amounts of the four ginsenosides and 20(S)-panaxadiol in the sweet taste variant were significantly higher than those in the bitter one. The significant difference between the sweet and bitter taste variants of GP was easily visualized in 3D-PCA score plots. The PCA loading plot also indicated the contributions among the four ginsenosides (Rd > Rb(3) > F(2) > Rb(1)) for distinguishing the two taste variants. This is the first report to describe the use of these two quantitative methods (UPLC-Q-TOF-MS and HPLC-ELSD) for the accurate authentication and quality control of GP.

Proteomic identification of proteins involved in the anticancer activities of oridonin in HepG2 cells.

Oridonin is the main bioactive constituent of the Chinese medicinal herb Isodon rubescens and has been shown to have anti-neoplastic effects against a number of cancers in vitro and in vivo. Here we report the proteomic identification of proteins involved in the anticancer properties of oridonin in hepatocarcinoma HepG2 cells. Cell viability assay showed that oridonin dose-dependently inhibited cell growth with an IC(50) of 41.77µM. Treatment with oridonin at 44µM for 24h induced apoptosis and G2/M cell cycle arrest, which were associated with nine differentially expressed proteins identified by proteomic analysis. The proteomic expression patterns of Hsp70.1, Sti1 and hnRNP-E1 were confirmed by quantitative real-time PCR and/or immunoblotting. Eight of the nine identified proteins are shown, for the first time, to be involved in the anticancer activities of oridonin. Up-regulation of Hsp70.1, STRAP, TCTP, Sti1 and PPase, as well as the down-regulation of hnRNP-E1 could be responsible for the apoptotic and G2/M-arresting effects of oridonin observed in this study. Up-regulation of HP1 beta and GlyRS might contribute to inhibitory effects of oridonin on telomerase and tyrosine kinase, respectively. These findings shed new insights into the molecular mechanisms underlying the anticancer properties of oridonin in liver cancer cells.

Oridonin induces G2/M cell cycle arrest and apoptosis through MAPK and p53 signaling pathways in HepG2 cells.

Oridonin, the main active constituent of Isodon rubescen, has antihepatocarcinoma activity in experimental and clinical settings. The aims of the study were to explore the anticancer effect of oridonin in HepG2 cells and to investigate the underlying mechanisms. Results showed that oridonin treatment for 24 or 48 h resulted in a marked decrease in cell viability time- and dose-dependently. IC50 values were determined as 38.86 microM and 24.90 microM for 24-h and 48-h treatments, respectively. Flow cytometric analysis showed that a 24-h treatment of 40 microM oridonin induced G2/M cell cycle arrest and apoptosis. Typical apoptotic nucleus alterations were observed with fluorescence microscope after DAPI staining. Immunoblot analysis demonstrated that oridonin treatment increased expression levels of p-JNK, p-p38, p-p53 and p21, elevated the level of cyclin B1/p-Cdc2 (Tyr15) complex, and inhibited the expression of p-ERK. Moreover, oridonin treatment activated caspase-9 and caspase-3. In conclusion, oridonin induced G2/M cell cycle arrest and apoptosis in HepG2 cells through MAPK and p53 pathways, which advances our understanding on the molecular mechanisms of oridonin in hepatocarcinoma management.

Liposomes modulate docetaxel-induced lipid oxidization and membrane damage in human hepatoma cells.

The aim of this study was to investigate the effect of liposomes on docetaxel-induced lipid oxidization and membrane damage in human hepatoma cells. Cytotoxicity of free docetaxel and docetaxel-containing liposomes was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium (MTT) assay in human hepatoma cell lines HepG2 and SMMC-7721. To the cell lines, blank liposomes prepared with soybean phosphatidylcholine (SPC), dimyristoylphosphocholine (DMPC), and dioleoylphosphocholine (DOPC) did not show any significant toxicity below a 0.02-mg/mL phospholipid concentration. On the other hand, free docetaxel showed IC(50) values of 9.13 x 10(-6) +/- 1.54 x 10(-5) and 1.58 x 10(-2) +/- 2.71 x 10(-2) mg/mL in HepG2 cells and SMMC-7721 cells, respectively, after of 24 hours of incubation. IC(50) values of docetaxel-encapsulating liposomes, measured in terms of total docetaxel concentration, were at least 1.5-fold higher than those of free docetaxel. SPC liposomes reduced cellular damage caused by free docetaxel, as evidenced by the attenuation of docetaxel-induced lactate dehydrogenase (LDH) leakage by over 11% after liposome encapsulation at each dosage. Docetaxel-induced oxidative membrane damage was monitored by the formation of the lipid peroxidation product, malondialdehyde (MDA), and the antioxidative property of SPC liposome was monitored by the suppression of superoxide dismutase (SOD). These data demonstrated that free docetaxel facilitated MDA formation and suppressed SOD, and that these membrane-damaging effects were reduced by SPC liposomes.

Anti-inflammatory and analgesic effects of the ethanol extract of Rosa multiflora Thunb. hips.

This study aimed to assess the anti-inflammatory and analgesic effects of Fructus Rosae Multiflorae (FRM, hips of Rosa multiflora Thunb.). FRM was extracted with 75% ethanol and the dried extract (FRME) was administered intragastrically (i.g.) at 100, 200 and 400mg/kg. The anti-inflammatory effect was evaluated in four experimental animal models and analgesic effect in two animal models. Pretreatment with a single dose of FRME produced significant dose-dependent anti-inflammatory effects on carrageenin-induced rat hind paw edema, xylene-induced mouse ear edema and acetic acid-induced mouse vascular permeation. In a 7-day study, daily administration of FRME suppressed cotton pellet-induced rat granuloma formation. Pretreatment with a single dose of FRME also produced dose-dependent anti-nociceptive effects in thermally- and chemically induced mouse pain models. In addition, a single dose of FRME at 2.4g/kg body weight (equivalent to 87.6g of dried hips per kg body weight) produced no observable acute toxicity in mice within seven days. These results demonstrate that FRME possesses anti-inflammatory and analgesic effects and has no obvious acute toxicity, which advanced our understanding of the folk use of FRM in treating various inflammatory disorders.