Hyperoside Suppresses Lipopolysaccharide-induced Inflammation and Apoptosis in Human Umbilical Vein Endothelial Cells

Finding the novel drug from the effective components of traditional Chinese herbal medicine is a hotspot of the modem pharmacological research. Hyperoside （HYP） belongs to flavonoid glycosides, and it has various properties, such as anti-inflammation, anti-spasm, anti-diuretic, antitussive, lowering blood pressure, and lowering cholesterol effects as well as protective effects for the cardiac and cerebral blood vessels. The purpose of this study was to investigate the effects of HYP on inflammatory and apoptotic responses in vascular endothelial cells stimulated by lipopolysaccharide （LPS） and further to identify the possible mechanisms underlying these effects. In our study, human umbilical vein endothelial cells （HUVECs） were stimulated with 1 Bg/mL LPS in the presence or absence of HYP （10, 20 and 50 μmol/L）. Our results indicated that HYP alone exerted no cytotoxicity on HUVECs, while it had an upregulatory effect on the viability of HUVECs induced by LPS in a dose-dependent manner; increased mRNA expression of IL-1β, IL-6, TNFα and iNOS induced by LPS was attenuated after treatment with HYP both in a dose- and time-dependent manner; LPS-induced HUVECs apoptosis and cleaved-caspase 8, 9, 3 were all significantly reduced by HYP. Furthermore, the possible pathway involved in apoptosis and inflammation by HYP was detected, and the results showed that when treated with HYP, LPS-induced mitochondrial membrane instability was significantly inhibited through up-regulation of Bcl-2 and down-regulation of Bax. Furthermore, the expression of TLR4 and the phosphorylation of IκBα and p65 in LPS-treated cells were blocked by HYP. Our results suggested that HYP treatment prevented HUVECs from LPS- induced inflammation and apoptosis responses, which might be mediated by inhibiting TLR4/ NFκB pathway.

Bear Bile Powder Inhibits Angiogenesis In Vivo and In Vitro

Objective： To evaluate the effect of bear bile powder （BBP） on angiogenesis, and investigate the underlying molecular mechanisms. Methods： A chick embryo chorioallantoic membrane （CAM） assay was used to evaluate the angiogensis in vivo. Human umbilical vein endothelial cells （HUVECs） were treated with 0, 0.25, 0.5, 0.75, and 1.0 mg/mL of BBP for 24, 48 and 72 h, respectively. The 3-（4, 5-dimethylthiazol-2-yl）-2,5- diphenyltetrazolium bromide assay was performed to determine the viability of HUVECs. Ceil cycle progression of HUVECs was examined by fluorescence-activated cell sorting （FACS） analysis with propidium iodide staining. HUVEC migration was determined by wound healing method. An ECMatrix gel system was used to evaluate the tube formation of HUVECs. The mRNA and protein expression of vascular endothelial growth factor （VEGF）-A in both HUVECs and HepG2 human cells were examined by reverse transcription-polymerase chain reaction and enzyme linked immunosorbent assay, respectively. ]Results： Compared with the untreated group, BBP inhibited angiogenesis in vivo in the CAM model （P〈0.01）. In addition, treatment with 0.25-1 mg/mL of BBP for 24, 48, or 72 h respectively reduced cell viability by 14%-27%, 29%-69% and 33%-91%, compared with the untreated control cells （P〈0.01）. Additionally, BBP inhibited the proliferation of HUVECs via blocking the cell cycle G1 to S progression, compared with the S phase of untreated cells 48.05% ± 5.00%, 0.25-0.75 mg/mL BBP reduced S phase to 40.38% ± 5.30%, 36.54 ±4.50% and 32.13 ± 3.50%, respectively （P〈0.05）. Moreover, BBP inhibited the migration and tube formation of HUVECs, compared with the tube length of untreated cells 100%± 12%, 0.25-0.75 mg/mL BBP reduced the tube length to 62% ± 9%, 43% ± 5% and 17% ± 3%, respectively （P〈0.01）. Furthermore, BBP treatment down-regulated the mRNA and protein expression levels of VEGF-A in both HepG2 cells and HUVECs. Conclusion： BBP could inhibit the angiogenesis by reducing VEGF-A expression, which may, in part, explain its anti-tumor activity.