ABSTRACT
AIM: To investigate the protective effects of preconditioning human umbilical vein endothelial cells (HUVECs) with Polygonum multiflorum stilbeneglycoside (PMS) under anoxia/reoxygenation (A/R), and the mechanism of protection. METHODS: Prior to A/R, HUVECs were incubated with PMS (0.6 x 10(-11), 1.2 x 10(-11), or 2.4 x 10(-11) mol/L) for 3 h. Cell injury was subsequently evaluated by measuring cell viability with an MTT assay and lactate dehydrogenase (LDH) release, whereas lipid peroxidation was assayed by measuring malondialdehyde (MDA) content. Antioxidant capacity was quantified by superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activity. Nitric oxide (NO) production was determined by nitrite accumulation. Endothelial NO synthase (eNOS) and inducible NOS (iNOS) protein expression was detected by Western blotting. Guanylate cyclase activity and cyclic GMP (cGMP) activity were assessed by an enzyme immunoassay kit. RESULTS: PMS incubation attenuated A/R-induced injury in a concentration-dependent manner, as evidenced by a decrease in LDH activity and an increase in cell viability. PMS exerted its protective effect by inhibiting the A/R-mediated elevation of MDA content, as well as by promoting the recovery of SOD and GSH-Px activities. Additionally, PMS incubation enhanced NO and cGMP formation by increasing iNOS expression and guanylate cyclase activity. The protective effects of PMS were markedly attenuated by NOS inhibitor L-NAME, soluble guanylate cyclase inhibitor ODQ or PKG inhibitor KT5823. CONCLUSION: PMS preincubation resulted in the enhancement of antioxidant activity and anti-lipid peroxidation. The NO/cGMP/cGMP-dependent protein kinase (PKG) signaling pathway was involved in the effect of PMS on HUVECs.
Subject(s)
Antioxidants/therapeutic use , Drugs, Chinese Herbal/therapeutic use , Endothelial Cells/drug effects , Glycosides/therapeutic use , Polygonum/chemistry , Reperfusion Injury/drug therapy , Stilbenes/therapeutic use , Cell Survival/drug effects , Cells, Cultured , Endothelial Cells/cytology , Endothelial Cells/metabolism , Glutathione Peroxidase/metabolism , Guanylate Cyclase/metabolism , Humans , L-Lactate Dehydrogenase/metabolism , Lipid Peroxidation/drug effects , Nitric Oxide/metabolism , Nitric Oxide Synthase/metabolism , Superoxide Dismutase/metabolism , Umbilical Veins/cytologyABSTRACT
AIM: To study the effect of Eupolyphaga Sinensis Walker(ESW) on red blood cell immune adherence (RCIA) and serum anticardiolipin antibody level in rat model of Yin-deficiency Huo-excess with chronic blood stasis. METHODS: The model was made by i.m. injection of dexamethasone (0.5 mg/kg) and adrenaline (0.84 mg/kg). The serum anti-cardiolipin antibodies (ACA) ACA-IgG, ACA-IgA and ACA-IgM and the plasma D-dimmer levels were measured by using enzyme-linked immunosorbent assay (ELISA). The body and organ weight of the rats were measured. RESULTS: For rats of Yin-deficiency Huo-excess, rosette rates of red blood cell C3b receptors (RBC-C3b RR) and red blood cell cancer (RBC-CaR) decreased, the serum ACA-IgG, ACA-IgA, ACA-IgM and the plasma D-dimmer levels markedly increased, body weight and the weight of spleen and thymus all decreased. ESW (5 mg/kg, 10 mg/kg) increased RBC-C3bRR and RBC-CaR, reduced serum ACA-IgG, ACA-IgA, ACA-IgM and plasma D-dimmer levels, and increased spleen weight of the rats. CONCLUSION: ESW can boost the immune function in rats of Yin-deficiency Huo-excess with chronic blood stasis.
