Effect of Buthus martensi Karsch on Aromatase Activity and Cytokine-Inducded NOS and NO Production in Osteoblasts and Leukaemic Cell Line FLG 29.1

Abstract

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.

Key Words :

• Aromatase
• Buthus martensi Karsch (BMK)
• Cytokine
• IL-1β
• NO
• NOS
• Osteoblast
• TNF-α