Gleditsia sinensis Thorn Extract Inhibits Proliferation and TNF-α-Induced MMP-9 Expression in Vascular Smooth Muscle Cells

Abstract

The thorns of Gleditsia sinensis, which are extensively used as a medicinal herb in Asian countries, have been reported to exert various pharmacological effects. However, the anti-atherogenic effect of Gleditsia sinensis thorns has never been investigated. In the present study, we investigated the role and effect of the ethanol extract of Gleditsia sinensis thorns (EEGS) on cultured vascular smooth muscle cells (VSMC). Treatment of VSMC with EEGS led to a significant decrease in cell growth by arresting cells in the G2/M-phase of the cell cycle, which was associated with up-regulated p21WAF1 levels and suppression of G2/M cell cycle regulators, cyclinB1, Cdc2 and Cdc25c. In addition, EEGS treatment led to the induction of extracellular signal-regulated kinase1/2 (ERK1/2), p38 MAPK, and JNK (c-Jun N-terminal kinases) activation. EEGS-induced p21WAF1 expression was blocked by treatment with the p38 MAPK-specific inhibitor SB203580. SB203580 also markedly recovered the inhibition of cell growth and decrease in cell cycle proteins in EEGS-treated VSMC. Moreover, EEGS inhibited matrix metalloproteinase-9 (MMP-9) expression induced by tumor necrosis factor-α (TNF-α) in VSMC. Finally, an electrophoresis mobility shift assay demonstrated that EEGS suppressed expression of transcription factor, nuclear factor kappaB (NF-κB) and activator protein-1 (AP-1), which are essential cis-elements for the MMP-9 promoter in TNF-α-treated VSMC. These results demonstrate that EEGS exerts a potent inhibitory effect on cell proliferation and MMP-9 expression in VSMC. These unexpected novel findings represent theoretical data for the preventive and therapeutic use of EEGS for the treatment of atherosclerosis disease.