In vitro simulated digestion and in vivo metabolism of chlorogenic acid dimer from Gynura procumbens (Lour.) Merr.: Enhanced antioxidant activity and different metabolites of blood and urine.

Gynura procumbens (Lour.) Merr. is an evergreen edible vine in southern China. The antioxidant activity and metabolites of chlorogenic acid dimer from Gynura procumbens (Lour.) Merr. were evaluated by the model of in vitro digestion and in vivo metabolomics approach, respectively. Moreover, metabolites of chlorogenic acid dimer in blood and urine of Sprague-Dawley rats were determined by HPLC-ESI-QTOF-MS/MS. In vitro digestion results suggested the antioxidant activity of the purified chlorogenic acid dimer was significantly enhanced after simulated digestion. Meanwhile, in vivo metabolism results showed that 7 and 20 new metabolites were observed in blood and urine, respectively, suggesting that hydrolysis along with methylation, glucuronidation and other reactions may all happen when the chlorogenic acid dimer entered the digestive and metabolic systems, which inducing and exhibiting various biological activities through metabolism. PRACTICAL APPLICATIONS: Gynura procumbens (Lour.) Merr. (GPM) is an evergreen edible vine with the effects of anticancer, anti-inflammatory, antiviral, depressurization, and antioxidation. As a health care vegetable, it is not usually eaten in daily life. Our current study shows that chlorogenic acid dimer extracted from GPM has a significant enhanced antioxidant ability after gastro-intestinal digestion in vitro, and their metabolites in vivo of urine is far more than that of blood, which may indicate that the chlorogenic acid dimer can be fully absorbed and decomposed through the gastro-intestinal digestion and metabolism. Thus, GPM could be used as a functional food ingredient for antioxidant enhancement to promote the economic value. The research also provides theoretical data for the intensive processing and utilization of GPM, as well as for the relative research on digestion and metabolism of edible plants.

The Octyl Ester of Ginsenoside Rh2 Induces Lysosomal Membrane Permeabilization via Bax Translocation.

Ginsenoside Rh2 is a potential pharmacologically active metabolite of ginseng. Previously, we have reported that an octyl ester derivative of ginsenoside Rh2 (Rh2-O), has been confirmed to possess higher bioavailability and anticancer effect than Rh2 in vitro. In order to better assess the possibility that Rh2-O could be used as an anticancer compound, the underlying mechanism was investigated in this study. The present results revealed that lysosomal destabilization was involved in the early stage of cell apoptosis in HepG2 cells induced by Rh2-O. Rh2-O could induce an early lysosomal membrane permeabilization with the release of lysosomal protease cathepsins to the cytosol in HepG2 cells. The Cat B inhibitor (leu) and Cat D inhibitor (pepA) inhibited Rh2-O-induced HepG2 apoptosis as well as tBid production and Δφm depolarization, indicating that lysosomal permeabilization occurred upstream of mitochondrial dysfunction. In addition, Rh2-O induced a significant increase in the protein levels of DRAM1 and Bax (p < 0.05) in lysosomes of HepG2 cells. Knockdown of Bax partially inhibited Rh2-O-induced Cat D release from lysosomes. Thus it was concluded that Rh2-O induced apoptosis of HepG2 cells through activation of the lysosomal-mitochondrial apoptotic pathway involving the translocation of Bax to the lysosome.

Esterification of Ginsenoside Rh2 Enhanced Its Cellular Uptake and Antitumor Activity in Human HepG2 Cells.

Our previous research had indicated that the octyl ester derivative of ginsenoside Rh2 (Rh2-O) might have a higher bioavailability than Rh2 in the Caco-2 cell line. The aim of this study was to investigate the cellular uptake and antitumor effects of Rh2-O in human HepG2 cells as well as its underlying mechanism compared with Rh2. Results showed that Rh2-O exhibited a higher cellular uptake (63.24%) than Rh2 (36.76%) when incubated with HepG2 cells for 24 h. Rh2-O possessed a dose- and time-dependent inhibitory effect against the proliferation of HepG2 cells. The IC50 value of Rh2-O for inhibition of HepG2 cell proliferation was 20.15 µM, which was roughly half the value of Rh2. Rh2-O induced apoptosis of HepG2 cells through a mitochondrial-mediated intrinsic pathway. In addition, the accumulation of ROS was detected in Rh2-O-treated HepG2 cells, which participated in the apoptosis of HepG2 cells. Conclusively, the findings above all suggested that Rh2-O as well as Rh2 inducing HepG2 cells apoptosis might involve similar mechanisms; however, Rh2-O had better antitumor activities than Rh2, probably due to its higher cellular uptake.

A ROS-mediated lysosomal-mitochondrial pathway is induced by ginsenoside Rh2 in hepatoma HepG2 cells.

Ginsenoside Rh2 (GRh2), isolated from Panax ginseng C. A. Meyer, has been proven as an anticancer compound both in vitro and in vivo. In the present study, we investigated the role of the lysosomes during the apoptosis of HepG2 cells induced by GRh2. The results showed that GRh2 significantly induced intracellular reactive oxygen species (ROS) generation in the HepG2 cells, which consequently resulted in early lysosomal membrane permeabilization with the release of cathepsin B (Cat B) to the cytosol. Western blot analysis showed that the released Cat B in the cytosol contributed to Bid cleavage. Subsequently mitochondrial damage was observed in the HepG2 cells. Interestingly, when the HepG2 cells were pre-treated with N-Acetyl-L-Cysteine (NAC) for 1 h, which inhibited ROS generation before being exposed to GRh2, the permeabilization of lysosomal membranes and the levels of Cat B in the cytosol were down-regulated. Moreover, mitochondrial damage was alleviated when the HepG2 cells were pre-treated with leupeptin (Leu). From the above results, it could be concluded that GRh2 induced apoptosis of the HepG2 cells through accumulation of ROS and activation of the lysosomal-mitochondrial apoptotic pathway involving the release of Cat B.