ABSTRACT
OBJECTIVE:To investigate the inhibitory effects mechanism of scallop skirt glycosaminoglycan(SS-GAG)on inju-ry in human umbilical vein endothelium cells (HUVEC). METHODS:In the test,there was a negative control group,a model group and the groups of SS-GAG at high,middle and low concentrations(mass concentrations of 200,100 and 50 mg/L respective-ly). The cells in latter 3 groups were cultured in SS-GAG at different mass concentrations for 12 h,and then in 50 μmol/L oxidized low-density lipoprotein(OX-LDL)for 24 h. MTT method was used to detect cell viability and the activity of lactic dehydrogenase (LDH),the flow cytometer to determine the level of reactive oxygen species (ROS),real-time fluorescence quantitative poly-merase chain reaction (RT-PCR) to detect mRNA expression of lectin-like oxidized low-density lipoprotein receptor 1 (LOX-1), and Western blot to detect NOX4 protein expression. RESULTS:Compared to the cells in the negative control group,those in the model group demonstrated lower viability,higher activity of LDH,higher level of ROS,and stronger expressions of LOX-1 mRNA and NOX4 protein. There was statistical significance (P<0.01). Compared to the cells in the model group,those in the groups of SS-GAG at high,middle and low concentrations showed higher viability,lower activity of LDH,lower level of ROS and weaker expressions of LOX-1 mRNA and NOX4 protein. There was statistical significance (P<0.01). CONCLUSIONS:SS-GAG can protect HUVEC to some degree by a mechanism which may be related to inhibiting ROS production via LOX-1/NOX4 pathway and relieving oxidative stress injury.
ABSTRACT
AIM To investigate if scallop (Placopecta magellanicus) skirt glycosaminoglycan (SS-GAG) inhibits the proliferation of vascular smooth muscle cell (VSMC) as heparin does so and to clarify its mechanism. METHODS The inhibitory effects of SS-GAG on the proliferation of rat thoracic aorta and abdominal aorta VSMC induced by fetal bovine serum (FBS) or basic fibroblast growth factor (bFGF) were determined by cell counting, crystal violet staining and MTT colorimetry. The effects of SS-GAG on the expression of proliferating cell nuclear antigen (PCNA) and platelet-derived growth factor (PDGF) in VSMC proliferation induced by bFGF were evaluated by immunohistochemical technique (LSAB method) and computer image analysis system. RESULTSSS-GAG exerted antagonistic effects on VSMC proliferation induced by 20% FBS and 50 μg·L-1 bFGF at concentrations ranging from 50 mg·L-1 to 200 mg·L-1 and repressed the increasing expression of PCNA and PDGF. CONCLUSION SS-GAG significantly inhibits the proliferation of VSMC, which may be carried out through repression of PDGF and PCNA expression.