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Objective:This study observed attenuating effect of hydroxysafflor yellow A(HSYA),an effective ingredient of aqueous extract of Carthamus tinctorius L,on lipopolysaccharide(LPS)-induced endothelium inflammatory injury.Methods:Eahy926 human endothelium cell(EC) line was used;thiazolyl blue tetrazolium bromide(MTT) test was assayed to observe the viability of EC;Luciferase reporter gene assay was applied to measure nuclear factor- κB(NF- κB) p65 subunit nuclear binding activity in EC;Western blot technology was used to monitor mitogen activated protein kinase(MAPKs) and NF- k B activation.Reverse transcription polymerase chain reaction(RT-PCR) method was applied to observe intercellular cell adhesion molecule-1(ICAM-1) and E-selectin mRNA level;EC surface ICAM-1 expression was measured with flow cytometry and leukocyte adhesion to EC was assayed with Rose Bengal spectrophotometry technology.Results:HSYA protected EC viability against LPS-induced injury(P<0.05).LPS-induced NF-κB p65 subunit DNA binding(P<0.01) and nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor α(I κ Bα) phosphorylation was inhibited by HSYA.HSYA attenuated LPS triggered ICAM-1 and E-selectin mRNA levels elevation and phosphorylation of p38 MAPK or c-Jun N-terminal kinase MAPK.HSYA also inhibited LPS-induced cell surface ICAM-1 protein expression(P<0.01) and leukocyte adhesion to EC(P<0.05).Conclusion:HSYA is effective to protect LPS-induced high expression of endothelium adhesive molecule and inflammatory signal transduction.
Objective: This study observed attenuating effect of hydroxysafflor yellow A (HSYA), an effective ingredient of aqueous extract of Carthamus tinctorius L, on lipopolysaccharide (LPS) -induced endothelium inflammatory injury. Methods: Eahy 926 human endothelium cell (EC) line was used; thiazolyl blue tetrazolium bromide (MTT) test was assayed to observe the viability of EC; Luciferase reporter gene assay was applied to measure nuclear factor-κB (p65-subunit nuclear binding activity in EC; Western blot technology was used to monitor mitogen activated protein kinase (MAPKs) and NF-k B activation. Reverse transcription polymerase chain reaction (RT-PCR) method was applied to observe intercellular cell adhesion molecule- 1 (ICAM- 1 expression was measured with flow cytometry and leukocyte adhesion to EC was assayed with Rose Bengal spectrophotometry technology. Results: HSYA protected EC viability against LPS-induced injury (P <0.05). LPS-induced NF-κB p65 su Bunit DNA binding (P <0.01) and nuclear factor of kappa light polypeptide gene enhancer in B-cell inhibitor α (I κ Bα) phosphorylation was inhibited by HSYA. HSYA attenuated LPS triggered ICAM-1 and E-selectin mRNA levels elevation and phosphorylation of p38 MAPK or c-Jun N-terminal kinase MAPK.HSYA also inhibited LPS-induced cell surface ICAM-1 protein expression (P <0.01) and leukocyte adhesion to EC (P <0.05) .Conclusion: HSYA was effective to protect LPS -induced high expression of endothelium adhesive molecule and inflammatory signal transduction.