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Effect of Sodium Selenite on Superoxide Anion-Induced Osteoblastic Differentiation of Vascular Smooth Muscle Cells and the Related Mechanisms

Author: XiaoRong
Tutor: LiuHongMei
School: Huazhong University of Science and Technology
Course: Inorganic Chemistry
Keywords: Sodium selenite Vascular smooth muscle cells Superoxide anion Vascular calcification
CLC: R363
Type: Master's thesis
Year: 2011
Downloads: 21
Quote: 0
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Abstract


Cardiovascular disease, including atherosclerosis、coronary artery disease,with highly morbidity and mortality,is a great threaten to human health. Atherosclerosis is frequently associated with vascular calcification. Vascular calcification is common characteristic of cardiovascular disease and bases on osteoblastic differentiation of vascular smooth muscle cells. Calcification of arteries is a complex and regulated process, some aspects of arterial calcification are similar to the biology of forming bone. Increasing evidences suggest that the supplementation of essential micronutrient selenium down-regulates the morbidity and mortality of cancer, suggesting selenium has positive effect against cancer. But whether selenium prevent cardiovascular disease remains unknown, as well as the possible signaling pathway. Selenium plays a crucial role in scavenging the oxygen radical. However, exogenous ROS（reactive oxygen species, ROS） including superoxide anion and hydrogen dioxide induce osteoblastic differentiation of VSMCs.The present research mimics the calcifying model of VSMCs in vitro, in accord with the relevant literatures. Xanthine oxidase（XO） and xanthine（X） system generates superoxide anion. The relevant trials try to reflect the effect of selenium on superoxide anion-induced osteoblastic differentiation of vascular smooth muscle cells.⒈The effect of different concentrations of X/XO on osteoblastic differentiation of VSMCs was investigated by the measurement of MTT assay on cell viability, the activity of alkaline phosphatase （ALP） and quantification of calcium deposition with Atomic Absorption Spectrometer（AAS）. The results indicated that low doses of X（μmol/L）/XO（mU/ml）（100/5,100/10） had no significant effect on osteoblastic differentiation of VSMCs; medium doses of X（μmol/L）/XO（mU/ml） （100/40） inhibits ALP acticity , reduced the quantification of calcium deposition and had no significant damage on cell survival; high doses of X（μmol/L）/XO（mU/ml） （100/80,100/100） resulted in increasing of ALP activity and matrix calcium deposition and marked damage on cell survival. ⒉The effect of Na2SeO3 on superoxide anion-induced osteoblastic differentiation of VSMCs. The calcifying cells were pretreatment with 0.1 or 0.05μmol/L Na2SeO3 for 24h and with PD98059, the special inhibitor of MAPKK, for 2h, before being exposed to X（μmol/L）/XO（mU/ml）（100/40 and 100/80）. Cell viability and the activity of alkaline phosphatase （ALP）, were determined. After treatment with X/XO for 3 days, The cytotoxicity of X/XO was significantly suppressed by pretreatment with 0.1 mM Na2SeO3 for 24 h, but the viability of calcifying VSMCs has no change after 7 days treatment. Data showed 0.1 and 0.05μmol/L Na2SeO3 reduced the ALP activitiy induced by superoxide anion, suggesting that Na2SeO3 had inhibition on superoxide anion-induced osteoblastic differentiation of VSMCs.⒊The potential mechanism of Na2SeO3 on superoxide anion-induced osteoblastic differentiation of VSMCs was researched. Cellular glutathion peroxidase（GPx） activities and intracellular ROS levels were determined. Western blotting was used to analyze the level of ERK phosphorylation. X/XO（100/40） decreased the GPx activity, which was increased significantly by pretreatment with Na2SeO3 for 24h. However, comparison with calcifying control group, intracellular ROS levels of X/XO group alone and X/XO group by pretreatment with Na2SeO3 increased, but Na2SeO3 had no inhibition on ROS levels induced by superoxide anion. The effect of X/XO on ERK phosphorylation in calcifying VSMCs were suppressed by Na2SeO3 pretreatment, which were inhibited remarkably by PD98059 for 2h, the special inhibitor of MAPKK in MAPK signaling pathway. Results above indicated that Na2SeO3 might suppress superoxide anion-induced osteoblastic differentiation of VSMCs through the MAPK transduction signaling pathway.

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