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Aim To study the intervention effect of tet- ramethylpyrazine(TMP) on human microvascular endothelial cells (HMECs) inflammatory response induced by activated complement alternative pathway and the possible molecular mechanisms. Methods HMECs were pretreated with different concentrations of TMP, and then exposed to the activated products of the complement alternative pathway which was prepared by cobra venom factor( CVF). The supernatant was removed and assayed for expression of the adhesion molecules (ICAM-1, VCAM-1 and E-selectin) and the inflammatory mediator(IL-6 and TNF-ot) by using ELISA reagent kits. The nucleus transcriptional activity of NF- kB was measured by the dual luciferase reporter assay ? system. Results The adhesion molecules, inflammato ry mediator and nucleus transcriptional activity of NF- kB increased after HMECs were exposed to the products of the activated complement alternative pathway. The up-regulation of ICAM-1, VCAM-1, E-selectin, IL-6, TNF-a and the nucleus transcriptional activity of NF-kB were inhibited by various concentrations of TMP in a dose-dependent manner. Conclusions TMP can effectively reduce inflammatory response of HMECs in-duced by the activated complement alternative pathway , and the mechanism may be highly related to inhibition of nucleus transcriptional activity of NF-kB.
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[ ABSTRACT] AIM:To determine the role of transcription factor Bach1 in the functions of human microvascular en-dothelial cells ( HMVECs ) .METHODS: Bach1 siRNA was transfected into HMVECs to knock down the expression of Bach1.In vitro endothelial cell tube formation assay in Matrigel culture was used as a surrogate assay for angiogenic poten-tial.Migration of HMVECs was determined by using Transwell chambers.Cell proliferation was measured by CCK-8 assay. Real-time PCR, Western blotting, and ELISA were employed to determine mRNA expression and protein level.Reporter as-say was performed to determine vascular endothelial growth factor ( VEGF) transcriptional activity.RESULTS:Knockdown of Bach1 expression in HMVECs led to an increase in the tube formation and increased endothelial cell migration ability, whereas it has little effect on cell proliferation.Bach1 silencing increased the mRNA and protein expression of heme oxygen-ase-1 (HO-1), and enhanced VEGF transcriptional activation, and mRNA and protein expression.CONCLUSION:Bach1 silencing increases HO-1 and VEGF expression, thus promoting the cell migration and tube formation of HMVECs, indicating that Bach1 is a repressor for angiogenesis.
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0.990).The intra-assay and inter-assay variation of the method was 3.10 % and 4.93 %,respectively.The all-trans ratinoic acid(ATRA) up-regulated t-PA mRNA expression in a dose-dependent manner(1.25~20.00 ?mol?L~(-1),P