ABSTRACT
Objective:To observe the effects of milrinone on LPS-induced inflammation and NF-κB activation of human coronary artery endothelial cells (HCAEC).Methods:HCAEC were cultured in vitro,and were divided into normal group,LPS group (1 μg/ml),milrinone 50 μmol/L group and milrinone 200 μmol/L group,the rear two groups were co-incubated with LPS for 24 h. qRT-PCR and ELISA was applied to detect IL-6,IL-8,TNF-α mRNA and protein,respectively.Western blot and immunofluorescence assay was used to observe the activation and translocation of NF-κB,respectively.Results: Compared with LPS group,the IL-6,IL-8, TNF-α mRNA and protein;the activation and translocation of NF-κB significantly inhibited after incubation with milrinone (P<0.05) in a dose dependent manner.Conclusion:Milrinone inhibited the release of inflammative cytokines induced by LPS of HCAEC in vitro, partly by inactivation of NF-κB.
ABSTRACT
<p><b>AIM</b>To determine the effects of azide methyl anthraquinone derivative (AMAD) on growth inhibition and inducing apoptosis of multidrug resistant (MDR) KBv200 cells and parental drug-sensitive KB cells.</p><p><b>METHODS</b>Cytotoxicity was determined by tetrazolium (MTF) assay. Reactive oxygen species (ROS) levels and mitochondrial membrane potential (deltapsi(m)) in cells were labeled with DCFH-DA and DiOC6 and tested by flow cytometry. Annexin V stain and DNA ladder were used to examine the apoptosis of KB and KBv200 cells induced by AMAD.</p><p><b>RESULTS</b>AMAD was shown to inhibit the growth of KB and KBv200 cells significantly in a concentration-dependent manner, with mean IC50 of 0.36 and 0.45 micromol x L(-1), respectively. The generation of ROS increased obviously after the cells were treated with AMAD for 12 h, up to the peak in 24 h, meanwhile the levels of deltapsi(m) were time-dependently decreased. DNA fragmentation appeared on the agarose gel. Annexin V stain showed AMAD induced apoptosis of KB and KBv200 cells also in a concentration-dependent manner.</p><p><b>CONCLUSION</b>AMAD showed inhibitory effect on both MDR KBv200 cells and parental drug-sensitive KB cells. The mechanism of action was associated with the increase of the cellular ROS level and the decrease of the mitochondrial membrane potential induced by AMAD, which result in cell apoptosis.</p>
Subject(s)
Humans , Anthraquinones , Chemistry , Pharmacology , Antineoplastic Agents , Chemistry , Pharmacology , Apoptosis , Cell Proliferation , Dose-Response Relationship, Drug , Drug Resistance, Multiple , Drug Resistance, Neoplasm , KB Cells , Mitochondria , Physiology , Molecular Structure , Mouth Floor , Mouth Neoplasms , Pathology , Reactive Oxygen Species , Metabolism , Vincristine , PharmacologyABSTRACT
<p><b>AIM</b>Annonaceous acetogenin 89-2 was obtained from atemoya plant. To investigate the effect of 89-2 on experimental chemotherapy against xenografts derived from multidrug resistant KBv200 cells and parental drug-sensitive KB cells.</p><p><b>METHODS</b>Cytotoxicity was determined by tetrazolium (MTT) assay. The models of KB and KBv200 xenografts in nude mice were established to investigate the effect of 89-2 on experimental chemotherapy against cancer in vivo. Mechanistic experiments were conducted to examine the function of P-gp by Fura 2-AM assay.</p><p><b>RESULTS</b>The compound 89-2 showed potent cytotoxicity in KBv200 and KB cells, and the mean IC50 of 89-2 to KBv200 and KB cells was 48.7 and 64.6 nmol.L-1, respectively. The IC50 of 89-2 to multidrug resistant (MDR) cells was similar to that to the parental drug-sensitive cells (P < 0.05). In the models of KBv200 and KB cell xenografts in nude mice, 89-2 (0.90 mg.kg-1, q2d x 6) exhibited 52.3% and 56.5% in inhibiting the growth of xenografts, respectively. The toxicity was endurable. The intracellular accumulation of Fura-2 in KBv200 cells increased to 1.66, 2.03, and 2.74-fold, respectively, by addition of 12.8, 64 and 320 nmol.L-1 of 89-2.</p><p><b>CONCLUSION</b>Both MDR KBv200 cells and parental drug-sensitive KB cells were sensitive to the treatment of 89-2 in vitro and in vivo. The mechanism of overcoming MDR was associated with the decrease of P-gp function MDR cells.</p>