Effect of the Active Fraction of Flos Daturae on IFN-γ Induced Apoptosis in Immortalized Human Epidermal Keratinocyte Cells (HaCAT)

ABSTRACT

The present study is to investigate the effect of the active fraction of Flos Daturae (AF-FD) (dried flower of Datura metel L.) on IFN-γ induced apoptosis in a human keratinocyte cell line (HaCAT), which is a widely accepted mimic model of psoriasis. Accordingly, we used MTT and LDH to assay cell viability, FCM and Hoechst 33342 staining to assay cell apoptosis, and Western blotting to assay the expression of keratin 17 (K17). Administration of IFN-γ (5-125 U/mL) decreased MTT reduction and increased LDH leakage in a dose-dependent manner. Twenty-five U/mL IFN-γ elevated apoptosis significantly. The experimental results showed that AF-FD at the concentrations of 25 and 50 μg/mL alleviated the IFN-γ-induced inhibition of MTT reduction by 12.58% and 17.51%, and decreased LDH leakage by 14.47% and 15.83%, respectively. Through FCM and Hoechst 33342 staining, it was found that AF-FD can lower IFN-γ induced apoptosis at the concentrations of 12.5-50 μg/mL. The administration of 25 μg/mL AF-FD attenuated the IFN-γ induced over-expression of K17 from 206.32% to 178.83%. The effect of AF-FD on K17 expression may play a key role in protecting HaCAT cells from apoptosis.