Oxidatively modified low-density lipoprotein (OxLDL) is present in atherosclerotic lesions and has been proposed to play an important role in atherogenesis. In the present study, in order to clarify the structure-binding activity relationship of Asp-hemolysin-related peptides to OxLDL, we investigated the interaction between Asphemolysin-related peptides consisting of 4 to 29 amino acid residues and OxLDL. The incubation of OxLDL with each Asp-hemolysin-related peptide resulted in the formation of an Asp-hemolysin/OxLDL complex. In particular, the tetrapeptide, YKDG (P-4), bound to OxLDL and inhibited the OxLDL-induced macrophage proliferation in a dose-dependent manner. Furthermore, we demonstrated that lysophosphatidylcholine (LysoPC) extracted from OxLDL inhibited the binding of P-21 to OxLDL in a dose-dependent manner and synthetic [ 14 C]LysoPC bound to P-21. We propose here that the YKDG region is one of the important sites for the binding of these peptides to OxLDL, and LysoPC as a typical lipid moiety of OxLDL is attributed to the binding of OxLDL to these peptides.
Macrophage-derived foam cells play an important role in atherosclerotic lesions. Oxidized low-density lipoprotein (OxLDL) induces macrophage proliferation via the specific uptake of lysophosphatidylcholine (LysoPC) of OxLDL by class A, type I and type II macrophage scavenger receptors. We have previously shown that Asp-hemolysin from Aspergillus fumigatus binds to LysoPC as a typical lipid moiety of OxLDL. This study investigated the effect of the Asp-hemolysin-related peptide (P-21), a synthetic peptide derived from a region of Asp-hemolysin that is rich in positive charges, on macrophage proliferation induced by OxLDL. Mouse peritoneal macrophages were used for proliferation study. OxLDL induced macrophage proliferation in an oxidation time-dependent manner, and P-21 inhibited OxLDL-induced macrophage proliferation in a dose-dependent manner. Furthermore, the binding analysis of P-21 to OxLDL by dissociation-enhanced lanthanide fluorometric immunoassay indicated that P-21 binds to OxLDL. These results indicate that P-21 inhibits the OxLDL-induced macrophage proliferation through binding of P-21 to OxLDL.
Lysophosphatidylcholine (LPC), formed during low-density lipoprotein (LDL) oxidation and located within atherosclerotic plaques, regulates a variety of cellular functions, some of which could be construed to promote atherosclerotic lesion development, including vascular muscle cell proliferation, monocyte attraction, and endothelial cell apoptosis. We have previously reported that the synthetic peptide derived from Asp-hemolysin, named P-21, inhibits oxidized LDL (OxLDL)-induced macrophage proliferation through binding of P-21 to OxLDL. In this study, to clarify the interaction between P-21 and LPC as a typical lipid moiety of OxLDL, we examined the influence of P-21 on LPC-induced apoptosis in human umbilical vein endothelial cells (HUVECs). Based on flow cytometric analysis, using annexine V-fluorescein isothiocianate and propidium iodide as probes to assess apoptosis, LPC induced the apoptosis of HUVECs, and P-21 significantly inhibited this activity by 82.4%. Furthermore, dissociation-enhanced lanthanide fluorometric immunoassay indicated that LPC inhibited the binding of P-21 to OxLDL in a dose-dependent manner. A 50% inhibition dose was estimated to be 4.65 m mM of LPC. These results suggest that P-21 inhibits LPC-induced HUVEC apoptosis through binding of P-21 to LPC.
The new desiccator system with measures for the prevention of dew drops and the processing of the formaldehyde (FA) gas discharged from the final desiccator was produced, and the FA removal rate for various adsorbents was examined. For the prevention of dew drops in the desiccator, a hygroscopic bottle containing silica gel was used next to the FA gas generator, and humidity was adjusted by adjusting the interval between the FA gas outlet (a) and the desiccant (b). The removal of the harmful FA gas discharged from the final desiccator (n=5) is an important in the environmental preservation. To solve this problem, the FA gas was passed through an oxidation bottle containing KMnO(4)-H(2)SO(4) solution, and it was possible to confirm the complete decomposition of the FA by increase of the CO(2) and elimination of the FA. For the determination of the FA concentration in the desiccator, 100 ml air was beforehand collected using a gas collector into a 100 ml vial bottle containing 2 ml distilled water, and 50 ml of air from each desiccator was injected using a glass syringe. This was left under a slightly reduced pressure for 20 min, and the FA concentration was determined by the AHMT method. The FA removal rate after 1 h for each adsorbent (0.5 g) was 50% or more for chitin, KIMCO and silica gel. The removal efficacy for activated carbon was higher for fine particles than for coarse particles, and a dose-response relationship was established.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.