endothelin-1 and 3-4-dihydroxyphenyllactic-acid

To investigate the effects of β-(3,4-dihydroxyphenyl)lactic acid on oxidative stress stimulated by high glucose levels in human peritoneal mesothelial cells (HPMCs) in vitro.. HPMCs were incubated with 100 mol/l glucose followed by 0.625-20 mg/ml β-(3,4-dihydroxyphenyl)lactic acid. Reactive oxygen species (ROS) were quantified by flow cytometry. Relative levels of fibronectin-1 (FN1), collagen-I α(1) (COL1A1), endothelin-1 (EDN1) and haem oxygenase-1 (HMOX1) mRNA and protein were quantified by real-time reverse transcription-polymerase chain reaction and Western blotting, respectively. Absolute levels of FN1 and COLIA1 were quantified by enzyme-linked immunosorbent assay.. β-(3,4-Dihydroxyphenyl)lactic acid significantly decreased ROS levels, and EDN1 mRNA and protein levels, in dose- and time-dependent manners. HMOX1 mRNA and protein levels were significantly increased by β-(3,4-dihydroxyphenyl)lactic acid in dose-dependent manners. COL1A1 and FN1 mRNA and protein levels were significantly decreased by β-(3,4-dihydroxyphenyl)lactic acid in dose- and time-dependent manners.. β-(3,4-Dihydroxyphenyl)lactic acid inhibited oxidative stress and reversed increases in FN1 and COLIA1 induced by high glucose levels in HPMCs. This may contribute to a protective role in peritoneal fibrosis.

Topics: Base Sequence; Blotting, Western; Cells, Cultured; Collagen Type I; DNA Primers; Dose-Response Relationship, Drug; Endothelin-1; Enzyme-Linked Immunosorbent Assay; Epithelial Cells; Fibronectins; Flow Cytometry; Glucose; Heme Oxygenase-1; Humans; Lactates; Oxidative Stress; Peritoneum; Reactive Oxygen Species; Real-Time Polymerase Chain Reaction

Danxiongfang (DF) is a new Chinese medicine formula used to treat atherosclerosis and vascular restenosis. The active ingredients in DF are danshensu (DSS), tanshinones (cryptotanshinone, CT) and ferulic acid (FA). The aim of present study was to evaluate the inhibitory effects of DF and its active ingredients on cell proliferation and protection against hydrogen peroxide (H(2)O(2))-induced injury in rat vascular smooth muscle cells (VSMC) in vitro.. VSMC proliferation was assayed by cell counting and measurement of cell viability using the 3-(4, 5-dimethylthiazol -2yl)-2, 5-diphenyltetrazolium bromide (MTT) method and protein content was measured by the Bradford method. The nitric oxide (NO) level was detected by an assay kit. The endothelin-1 (ET-1) level was measured by ELISA. The protective effects of DF and its active ingredients on H(2)O(2)-induced cell injury was evaluated in terms of cell viability (MTT assay), superoxide dismutase (SOD) activity and malondialdehyde (MDA) levels. Hydroxyl free radicals generated by the Fenton reaction was detected with the spin-trapping technique on an electron spin resonance spectrometer.. The results suggest that DSS, CT, FA and DF inhibited VSMC proliferation by increasing the NO level and decreasing the ET-1 content. In rat VSMCs exposed to H(2)O(2), FA, DSS, CT and the six formulations of DF increased cell viability and SOD activity, and reduced the levels of MDA and hydroxyl free radicals. These effects of FA, DSS and CT occurred in a dose-dependent manner. Of the six formulas, DF 4 and DF 5 had the more significant activities. The effects of DF were much greater than those of the individual ingredients, even though the concentrations of these ingredients in the DF formulas were much lower than the doses of the individual ingredients used in each study, indicating markedly synergistic effects of DSS, CT and FA in DF on rat VSMCs.. these findings provide a pharmacological foundation for the clinical use of DF in the prevention and treatment of hyperlipidemia and atherosclerosis relevant to endothelial cell proliferation and damage.

Topics: Animals; Antioxidants; Aorta; Cardiotonic Agents; Cell Proliferation; Cell Survival; Cells, Cultured; Coumaric Acids; Dose-Response Relationship, Drug; Drug Synergism; Drugs, Chinese Herbal; Endothelin-1; Hydrogen Peroxide; Hydroxyl Radical; Lactates; Male; Malondialdehyde; Muscle, Smooth, Vascular; Myocytes, Smooth Muscle; Nitric Oxide; Phenanthrenes; Rats; Rats, Sprague-Dawley; Superoxide Dismutase