| Background Atherosclerosis(AS) is a disease that mainly involves large and medium arteries. It is characterized by a series of formation of lipid, fatty steaks, atheromatous plaque, plaque rupture hemorrhage and thrombosis,which lead to chronic vascular stenosis and even acute blocking, thus threatening people’s life. With the increasing morbidity of AS and the more and more studies about it by scientists, its pathogenesis are also varied, but two of them are widely accepted, inflammatory theory and endothelial injury theory. It is said that inflammatory factors and immune response play a vital role in the pathogenesis and development of AS.Chinese yam is not only edible but also used as medicine. It has been a cheap and fine supplement since ancient times as it is rich in nutrition. Drug products with dioscorease rhizome extract(DRE) as the main ingredients have been used in clinical for years, mainly in the prevention and treatment of AS related diseases, such as hyperlipidemia and coronary heart disease. However, as the lack of large clinical trials, animal and basic experiments, and the molecular mechanism study, the mechanism of how DRE protects AS is not yet to be investigated.PEST-containing nuclear protein(PCNP), a newly discovered protein located in nucleus and being rich in PEST sequence, is a sort of hydrophilic protein with short half-life and is relatively unstable. Because it is NIRF substrate, being ubiquitin by NIRF easily, and then it will be degraded by protease. Studies showed that PCNP participated in the regulation of cell cycle and the development and progression of kinds of tumor, and it also may play a role of regulation in rheumatoid arthritis. Tumor necrosis factor-α-induced protein 8-like 2(TIPE2) was also a newly discovered protein that regulated cell apoptosis, negatively regulated innate immunity and acquired immunity and participated in the maintenance of body’s immune homeostasis. As the immunosuppression after bacteria infection, the inflammatory response might be aggravated. Meanwhile, TIPE2 was also found to be involved in the development and progression of varieties of tumors. Recent studies suggest that TIPE2 expressed in macrophages inhibite the process of atherosclerosis by negative regulation of immune response.The project is to analyze the relationship between DRE and AS by the study of PCNP and TIPR2 expression in human coronary artery endothelial cells(HCAEC) and human aortic endothelial cell(HAEC), and also the effects of DRE on the expression of PCNP and TIPR2 in HCAEC and HAEC, to provide theoretical basis of the development of new drugs.Objective The project is to analyze the relationship between DRE and AS by the study of PCNP and TIPR2 expression in HCAEC and HAEC.Methods DRE was dissolved in DMEM culture media at the concentration of 10mg/ml as mother solution and was accordingly diluted at the concentration of 1μg/ml,10μg/ml, 100μg/ml, separately. Then cells HCAEC and HAEC were incubated with DRE at the above concentration for 24 h, 48 h, 72 h to study the effect of DRE on the cells growth by MTT, the gene expression of PCNP and TIPE2 by RT-PCR and the protein expression of PCNP and TIPE2 by Western Blotting(WB).Results DRE did not significantly affect the growth of HCAEC and HAEC evaluated by MTT and the expression of PCNP detected by WB and RT-PCR, while DRE positively regulated the expression of TIPE2 both on protein and gene levels.Conclusions 1. DRE did not significantly affect the growth of HCAEC and HAEC; 2. DRE and incubation time did not significantly affect the expression of PCNP, and there was no difference of the PCNP expression in HCAEC and HAEC. 3. DRE and the expression of TIPE2 were positively correlated, but no significant difference was found in the TIPE2 expression in different incubation time or the two kinds of cells. |
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