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Cell model of uric acid on vascular endothelial function and related research establishment

Author: WangZuo
Tutor: ZengXueJun
School: Beijing Union Medical College
Course: Internal Medicine
Keywords: Human Umbilical Endothelial Cell Uric Acid endothelial funcion oxidased LDL endothelial NO synthesase ICAM-1
CLC: R589.7
Type: Master's thesis
Year: 2011
Downloads: 96
Quote: 1
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Abstract


Backgrounds As the living conditions were improving, the prevalence of hyperuricemia increased markedly around the world. More and more epidemiological studies suggested that hyperuricemia was related with cardiovascular diseases, but it is not clear yet whether the increase of serum uric acid levels is an independent risk factor for them. Endothelial dysfunction was considered to play the key role for cardiovascular diseases. However, because of using different way to form cells model, there are still lots of debates regarding the effects of uric acid on vascular endothelial dysfunction.Objectives 1) To set up an HUVEC model in vitro which is suitable for the experiment to illustrate the effects of uric acid on vascular endothelial function.2) To explore the effects of uric acid alone or correlated with ox-LDL on the vascular endothelial function and their possible pathogenesis in vitro studies.Contents&Methods 1)To set up a suitable vascular endothelial cells model in vitro: HUVECs cultured in different medium were observed the morphous and their growth. Then, eNOS mRNA expression of HUVECs were determined by real-time PCR after they incubated with different conceration of ox-LDL at 24h.2) To explore the effects of uric acid on endothelial function by using the model determined by the part one: Real-time PCR was used to test the relative gene expression of eNOS and ICAM-1 mRNA in HUVEC stimulated by uric acid alone or uric acid with ox-LDL of stratified concentrations. Western blotting was also used to evaluate eNOS and ICAM-1 protein levels to observe the effects of uric acid alone. NO and s-ICAM contents in medium were detected by Enzymic method or ELISA in the meantime.Results 1)HUVEC cultured in ECM medium or 2:1 medium growed fast with the peak growth within 24h and lasted untill 72h. HUVEC cultured in 1:1 medium growed appropriately according with the growth of endothelial cell in vivo.50μg/ml ox-LDL downregulated eNOS expression on HUVEC in all medium; 20μg/ml ox-LDL downregulated the expression of eNOS on HUVEC cultured in 1:1 or 2:1 medium; 10μg/ml ox-LDL decreased eNOS expression just in 1:1 medium.2) High levels of uric acid(10、15 and 20 mg/dl) alone or with ox-LDL can decreased the expression of eNOS and the production of NO significantly in a time-dependent mode. High levels of uric acid stimulated HUVEC alone also upregulated the expression of ICAM-1. However, with ox-LDL, uric acid downregulated the expression of ICAM-1 mRNA. Uric acid of 5 mg/dl could increase the expression of eNOS and decrease s-ICAM in medium without ox-LDL. But along with ox-LDL, uric acid also downregulated the expression of eNOS just with the concentration of 5 mg/dl.Conclusion 1) HUVEC cultivated in medium, which composed by ECM and DMEM with the ratio of 1:1, was a appropriate model in vitro to examine the effects of uric acid on HUVEC.2)High levels of uric acid could induce the endothelial dysfunction, but uric acid in physiological concentration may protect the endothelial cells in healthy people. However, with high levels of ox-LDL existence, any levels of uric acid maydamaged endothelial function.

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CLC: > Medicine, health > Internal Medicine > Endocrine diseases and metabolic diseases > Metabolic diseases > Metabolic disorder of purine ( Purine )
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