Inhibition By Antilipoxygenase Drugs Of Cellular Chemiluminescence In Silica Activated Phagocytic Cells -- Alveolar Macrophages, Human Neutrophils And Human Leukemia (HL-60) Cells+

Since it has been demonstrated that silica caused phagocytic cells to activate lipoxygenase metabolism of unsaturated fatty acids we thought selective inhibition of this metabolic pathway might be helpful in treatment of silicosis. It is known that lipoxygenase metabolism plays a role in the pathogenesis of a variety of inflammatory and hypersensitivity conditions. Further, it is believed that the alveolar phagocytic cells which engulf silica begin the inflammatory cycle which produces the metabolic products, cytokines and growth factors which causes fibrosis in the lung. There have been indications that drugs which inhibit lipoxygenase can inhibit fibrosis from other causes. Previously, we developed a method which detects the activity of lipoxygenase metabolism by measuring free radical oxidation via light production or chemiluminescence (CL). We decided to apply this method to phagocytic cells that would be found in the lung in the presence of silica with and without newly developed inhibitors of the lipoxygenase reaction. We used drugs which inhibit lipoxygenase directly including AA861 (TAKEDA), A63162 (Abbott), and an inhibitor that scavenges free radicals produced from the lipoxygenase reaction, nordihydroguairetic acid. In addition we used an indirect inhibitor, tetrandrine. We found drug inhibition of silica activated CL from alveolar macrophages, neutrophils and human leukemia (HL-60) cells differentiated with DMSO. Nordihydroguairetic acid 2x10-5M produced almost complete inhibition in all three cell types. In human leukemia cells (HL-60) A63162 and tetrandrine were equally inhibitory at 42% while in human neutrophils A63162 and tetrandrine were less active 22 and 10% respectively. In macrophages A63162 and AA861 were less than 50% inhibitory to cellular CL at 2xl0-5M. Direct assay of tetrandrine (2x10-5M) with sodium arachidonate and soybean lipoxygenase reveals no inhibition. All antilipoxygenase drugs regardless of mechanism were inhibitory to cellular chemiluminescence from all three types of phagocytic cells. These drugs alone or in mixtures could prove useful in treatment of silicosis.