第一部 紅花萃取物與單一成分CTF-4-7C-5抑制FAS-death receptor途徑和活化IGF-1R以防止LPS誘導的H9c2細胞凋亡第二部分 紅花萃取物與單一成分CTF-4-7C-5抑制IGF-IIR途徑和活化IGF-1R以防止LPS誘導的H9c2細胞肥大

Abstract

Carthamus tinctorius is a traditional Chinese medical herb that has been used as a treatment for cardiovascular disease. The present study was performed to investigate the effect of Carthamus tinctorius extract (CTF) or the CTF pure compound (CTF-4-7C-5), on rat myocardium cell line H9c2 and the possible molecular mechanism. H9c2 cells were treated with LPS (2 μg/ml) for 12 h, and subsequently treated with CTF (1-25 μg/ml) or CTF-4-7C-5 (0.1-1μM). The incubation continued for another 24 h and the cells were analyzed with MTT assay, cell morphological changes assays, western blot analysis, flow cytometry, TUNEL, JC-1 staining and siRNA transfection assays. CTF or CTF-4-7C-5 increased H9c2 cell viability in a dose-dependent manner. Significant increases of TUNEL-positive cells, TNF-α, FAS-L, FAS, FADD, caspase-3, 8, 9 levels, cytosolic cytochrome c, pro-apoptotic protein markers, instability of mitochondria membrane potential, were observed in LPS-treated H9c2 cells. Unexpectedly, these LPS-enhanced effects were dose-dependently reduced by CTF at all doses, and 25 μg/ml was the most effective. It was also found that the survival pathway may involve anti-apoptotic effect of CTF on LPS-treated H9c2 cells. IGF-IR siRNA or inhibitors reversed the CTF effects, confirming that CTF activate IGF-1R to attenuate LPS-induced H9c2 cell apoptosis. The current findings indicate that CTF and compound CTF-4-7C-5 inhibit FAS-death receptor pathway and activate Insulin-like growth factor-I receptor pathway to reduce LPS-induced H9c2 cardiomyoblast cell apoptosis.