Involvement of NF-kappaB activation in the apoptosis induced by extracellular adenosine in human hepatocellular carcinoma HepG2 cells.

Adenosine can exhibit cytotoxic activity in vivo and in vitro, though its mechanisms are still uncertain. In this study, we investigated the adenosine-mediated apoptotic signaling pathway and the role of NF-kappaB in human hepatocellular carcinoma HepG2 cells. HepG2 cells were treated with different concentrations of adenosine for 12-48 h, and the effect of adenosine on cell proliferation was evaluated by MTT assay. The cytotoxicity of adenosine alone or in combination with an NF-kappaB inhibitor, pyrrolidine dithiocarbamate (PDTC), was also evaluated by MTT assay and the mode of cell death was detected by Hoechst 33342 staining. Cell cycle progress was performed by flow cytometry with PI staining. The protein expressions of Bcl-2, p53, NF-kappaB subunit p65, and caspase-3 were assayed by Western blot. Caspase-3 activity was measured by spectrophotomteric assay. The results showed that adenosine significantly reduced the viability of HepG2 cells in a dose- and time-dependent manner, with IC 50 (24 and 48 h) of 2.52 and 1.89 mmol x L(-1), respectively. The apoptotic index (percentage of sub-G1 phase) of HepG2 cells in adenosine treatment alone for 12 and 24 h or in combination with PDTC were 8.30%, 22.32% and 20.18%, 30.89%, respectively. All of them were higher than that in the control group (0.81%, p < 0.01). The characteristic changes of cell apoptosis (chromatin condensation and sub-G1 peak) were observed under fluorescent microscopy and flow cytometry. We also found that the apoptotic process triggered by adenosine was involved in G0-G1 cell-cycle arrest, enhanced the activity of caspase-3, upregulated p53 and NF-kappaB p65 expression, and downregulated Bcl-2 expression. Inhibition of NF-kappaB by PDTC decreased NF-kappaB p65 expression, enhanced cell apoptosis ratio, and increased caspase-3 activity. NF-kappaB may play an anti-apoptosis role in adenosine-induced HepG2 cytotoxicity.

A new saponin from Koelreuteria paniculata / 药学学报

To investigate the chemical constituents of seeds of Koelreuteria paniculata Laxm. , components were separated by means of solvent extraction and chromatography on C18, the structure of compound was determined by spectral analysis and chemical evidences. One saponin was obtained and identified as 28-O-isopentyryl-3beta, 16alpha, 22beta, 28-tetrahydroxyl-oleanane-3-O-[ alpha-L-rhamnopyranosyl- ( 1-->3) -betaD-galactopyranosyl-( 1--->4' )]-3betaD-galacturonopyranoside, named paniculata saponin C. This saponin is a new compound.

Establishment of Animal Model of Coronary Heart Disease with the Blending of Phlegm and Blood Stasis / 广州中医药大学学报

【Objective】To explore a method for the establishment of animal model of coronary heart disease(CHD) with the blending of phlegm and blood stasis.【Methods】Methods of TCM-WM combination(traditional Chinese medicine integrated with western medicine) and disease-syndrome combination were used to establish a model.The model was evaluated by the changes of symptoms,signs and laboratory indexes and by the effect of blood-stasis-removing and phlegm-resolving herbs on the model.Sixty Wistar rats were randomized into normal group,model group,low-dose herbal medicine group and high-dose herbal medicine group.The normal group was fed with routine forage and normal saline,and the other three groups with high-fat forage and fat milk(10mL?kg~(-1)?d~(-1)).Low-and high-dose herbal medicine groups were given Xuefu Zhuyu Decoction combined with Gualou Xiebai Banxia Decoction in the dosages of 12 and 24 10g?kg~(-1)?d~(-1)respectively by gastric gavage,and the model group given the same volume of normal saline for 7 consecutive weeks.Coronary artery ligation was performed in the 35~(th) day.The changes of electrocardiogram(ECG) were observed before and after the modeling,and blood lipid,myocardial enzymes and hemorrheological parameters were determined.The heart weight was examined to calculate heart index.【Results】ECG in the model group indicated severe myocardial ischemia,which may be relieved by herbal medicine.Serum contents of cholesteril,tryglyceride and creatine kinase(CK),whole blood viscosity,hematocrit and heart index were obviously increased in the model group,while the above indexes decreased to different degrees in herbal medicine groups (P