Effect Of Hsya On Lps-elicited Expression Of Mediators Of Inflammation Included Inos In Vascular Endothelial Cells

The study is aimed to investigate the effect of hydroxysafflor yellow A (HSYA) on lipopolysaccharide (LPS)-elicited the expressions of mediators of inflammation included inducible nitric oxide synthase (iNOS) in vascular endothelia1 cell line HUVECs.The study includes two parts:1. The change of expressions for mediators of inflammation in HUVECs cell line treated by LPS and HSYA. Methods: The HUVECs cell line was treated by different concentration of LPS (0.01 mg/L,0.1 mg/L and 1 mg/L) for 24 h. The mRNA of TNFα, IL-6, IL-8 and iNOS was detected by RT-PCR. Then the HUVECs cell line was treated by 1 mM/L HSYA for 2 h, and added 1 mg/L LPS. 24 h later, the mRNA of TNFα, IL-6 and iNOS was detected by RT-PCR. Results: The expression of IL-8 had no distinctive change with different concentration of LPS, while with higher concentration of LPS, the expression of TNFα, IL-6 and iNOS was increased. The expressions of TNFα, IL-6 and iNOS mRNA have significant change with 1 mg/L LPS. When the HUVECs cell line was treated by both 1 mg/L LPS and 1 mM/L HSYA for 24 h, the expression of TNFα, IL-6 mRNA had no significant change, while the expression of iNOS mRNA was decreased compared with the cells treated with 1 mg/L LPS. Conclusion: The HUVECs cell line has been cultivated successfully. Human umbilical vein endothelial cell damage model has been constructed successfully by 1mg/L LPS. INOS has been chosen for further study.2. To investigate the effect of HSYA on LPS-elicited iNOS expression in vascular endothelia1 cell line HUVECs.Methods: Human umbilical vein endothelial cells (HUVECs ) were induced by 1 mg/L LPS and HSYA(0.01 mmol/L,0.1 mmol/L,1 mmol/L). Cell survival rate was measured by MTT assay. NO level was detected by nitrate reductase method. The expression of iNOS was detected by RT-PCR and western blotting. Results: There was no significant change of LPS-elicited iN0S expression with 0.01 mmol/L and 0.1 mmol/L HSYA, while 1 mmol/L HSYA could obviously inhibited the expression of iNOS induced by LPS.Conclusion: HSYA can inhibit LPS-elicited iN0S expression and NO production induced by LPS in HUVECs cells, which may be used in the prevention and treatment of vascular inflammation diseases.