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Brefeldin A for 6 h, the relaxation induced by Bradykinin was completely abolished in the presence of 10mM Indomethacin and 30 μM L-NOARG. Treatment with 20 μg/mL Brefeldin A at concentrations between 1 nM and 1 mM abolished Bradykinin-induced [Ca2+]i reduction. Brefeldin A acts on endothelial cells and has no effect on Bradykinin or Substance P-induced [Ca2+]i enhancement. Brefeldin A did not affect the spontaneous phospholipid-dependent binding of GTPS to myr-rARF1, but completely abolished the catalytic exchange of retinal isotonic extract (RIE), and the semi-inhibitory concentration of 2 μM Brefeldin A was 2 μM. Brefeldin A inhibits multiple membrane transport pathways. Brefeldin A acts on the Golgi membrane or brain cell plasm to inhibit guanine nucleotide exchange activity specific to ADP-ribosylation factor. Complete inhibition of Brefeldin A indicates that the retinal extract contains ARF-specific guanine nucleotide exchange factors. Brefeldin A only partially inhibits ADP-ribosylation factor (ARFs) -released retinal isotonic extract (RIE) catalyzed GTPS, even at concentrations up to 300 μM. Brefeldin A induces the fusion of the Golgi apparatus with ER. Brefeldin A abolished the inhibitory effect of the CERT inhibitor HPA-12. Brefeldin A treats CHO cells to increase sphingomyelin synthesis by two to three times. In addition to B-CLL cells, Brefeldin A also causes apoptosis in multiple myeloma (U266, NCI-H929), JURKAT, HeLa cells, leukemia (HL60, K562, BJAB), colon (HT-29), and prostate, and adenoid cystic tumor cells. HF4.9 and HF28RA cells were treated with 25 ng/mL Brefeldin A to completely inhibit cell growth, while high doses of Brefeldin A (75 ng/mL) were required to completely inhibit the growth of HF1A3 cells. 50-75 ng/mL Brefeldin A treated HF1A3, HF4.9 and HF28RA cells, inhibited cell proliferation within 24 hours in a dose-dependent manner, and the penetration of 3H-thoracine was almost completely stopped when treated with 50 ng/ml. HF1A3, HF4.9, and HF28RA cells were inhibited by 26%, 76%, 87%, and 75%, 87%, respectively, when treated with 75 ng/mL. In YO-PRO 1/PI assay, Brefeldin A induced cell death in a dose-dependent manner.
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