Effect of Buthus martensi Karsch on aromatase activity and cytokine-inducded NOS and NO production in osteoblasts and leukaemic cell line FLG 29.1

Among the different scorpion species, Buthus martensi Karsch, a widely distributed scorpion species in Asia especially in Korea, has received a lot of attention. Indeed, over the past decade, more than 70 different peptides, toxins, or homologues have been isolated. It may prove a valuable resource for identifying potential anti-inflammatory and analgesic drugs. The recent observation has suggested that the aromatase is a possible local modulator of bone remodeling in osteoarthritis and osteoporosis. In the present study, therefore, the effect of Buthus martensi Karsch (BMK) extract, traditional immunosuppressive Korean aqua-acupuncture water, on the bone function of human osteoblastic cells was studied. To provide insights into the effect of BMK on aromatase activity in bone-derived cells, we examined the human leukaemic cell line FLG 29.1, which is induced to differentiate toward the osteoclastic phenotype by 12-O-tetradecanoyl-phorbol-13-acetate (TPA) and transforming growth factor (TGF)-β1, and the primary first-passage osteoblastic cells (hOB). Gene expression of the aromatase was not affected by Buthus martensi Karsch in FLG 29.1 and hOB cells. However, enzyme activity was stimulated in a time-dependent fashion by 10.0 μg/ml BMK and by either 1-50 nM TPA or 0.01-0.5 ng/ml TGF-β1, with maximal responses after 2-3 hr exposure. On the other hand, BMK strongly inhibited interleukin-1β (IL-1β)- and tumor necrosis factor (TNF)α-induced Nitricoxide (NO) synthase expression with little effect on constitutive NO synthase expression. BMK extracts (10 μg/ml) inhibited cytokine-induced iNOS and nNOS expression. BMK (10 μg/ml) did not affect the ecNOS expression, indicating the extracts are not working on the constitutive NOS expression. BMK strongly inhibited the cytokine-induced NO production (p < 0.01). BMK also showed significant inhibition on NO production in both induced by TNF-α+IL-1β. NO donors, sodium nitroprusside, and NONOate dose-dependently elevated alkaline phosphatase activity. These results suggest that NO directly facilitates osteoblastic differentiation. This result also suggests that BMK is effective for bone resorptive action in bone cells.