JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES) ›› 2015, Vol. 53 ›› Issue (11): 10-15.doi: 10.6040/j.issn.1671-7554.0.2015.255

• Preclinical Medicine • Previous Articles     Next Articles

Effects of trans-3,5,4'-trimethoxystilbene on the expressions of NO, ICAM-1 and NF-κB in human umbilical vein endothelial cells induced by lipopolysaccharide in vitro

FU Haiyan1, HU Zhansheng1, DU Hongyang2   

  1. 1. ICU, the First Affiliated Hospital of Liaoning Medical University, Jinzhou 121001, Liaoning, China;
    2. Department of Dermatology, the First Affiliated Hospital of Liaoning Medical University, Jinzhou 121001, Liaoning, China
  • Received:2015-03-10 Online:2015-11-10 Published:2015-11-10

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Abstract: Objective To explore the effects of trans-3,5,4'-trimethoxystilbene (TMS) on the expressions of NO, intercellular adhesion molecule-1 (ICAM-1) and nuclear factor-κB (NF-κB) in human umbilical vein endothelial cells (HUVEs) induced by lipopolysaccharide (LPS) in vitro. Methods The cell viabilities influenced by the different concentrations of TMS were assessed by CCK-8 assay. The cells were divided into the control group (CON group), LPS group, low-concentration TMS plus LPS group, medium-concentration TMS plus LPS group, high-concentration TMS plus LPS group and ammonium pyrrolidine dithiocarbamate (PDTC) plus LPS group. HUVEs were pretreated with the different concentrations of TMS and 10 μmol/L PDTC, and then were stimulated with 0.1 μg/mL LPS. After incubation, the level of NO was determined by Griess assay. The mRNA expressions of ICAM-1 and NF-κB p65 were detected by Real-time PCR, the protein expressions of ICAM-1, NF-κB p65 and IκBα by Western blotting assay, and the protein expressions of ICAM-1 and NF-κB p65 by immunocytochemetry assay. Results There was little effect of low-concentration TMS (5 or 10 μmol/L) on the cell viability, but the cell viability decreased significantly when treated with high-concentration TMS (50 or 100 μmol/L) in time- or concentration-dependent manners. Griess results showed that the level of NO in the low-, medium- and high-concentration TMS plus LPS groups and PDTC plus LPS group decreased compared with that in LPS group (P<0.05). The results of Real-time PCR and Western blotting showed that compared with LPS group and CON group, there were significant difference of mRNA and protein expressions of ICAM-1 and NF-κB p65 in medium-concentration TMS plus LPS group and PDTC plus LPS group (P<0.05; P<0.01). Furthermore, there were nuclear expressions of NF-κB p65 protein in medium-concentration TMS plus LPS group, PDTC plus LPS group and LPS group except for CON group. The protein expression of IκBα decreased significantly in LPS group compared with that in CON group (P<0.01), and those in medium-concentration TMS plus LPS group and PDTC plus LPS group were lower than those in CON group (P<0.05). Immunofluorescence assay showed there were hyperfluorescence of ICAM-1 in LPS group but weakened fluorescence in medium-concentration TMS plus LPS group and PDTC plus LPS group. There were no fluorescence of NF-κB p65 in CON group, cell nucleus hyperfluorescence in LPS group, and weakened fluorescence in medium-concentration TMS plus LPS group and PDTC plus LPS group with no cell nucleus expression. Conclusion TMS can inhibit LPS-mediated NO, ICAM-1 and NF-κB p65 expressions in HUVEs in vitro, and the inhibition of ICAM-1 may be affected through NF-κB cell pathway.

Key words: Intercellular adhesion molecule-1, Trans-3,5,4'-trimethoxystilbene, Lipopolysaccharide, Vein endothelial cells, NO, Nuclear factor-κB

CLC Number: 

  • R965.1
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