ABSTRACT
To investigate the effect of schisantherin A on liver sinusoid endothelial cell function and angiogenesis. Different dosages (0-40 μmol•L⁻¹) of schisantherin A were incubated 24 h with SK-HEP-1 cells, and the toxicity of SK-HEP-1 cells was assayed by MTT method. The proliferation of SK-HEP-1 cells were induced by the vascular endothelial growth factor (VEGF), with receptor tyrosine kinase inhibitor sorafenib as the control, at the same time, set up the control group, 2, 20 μmol•L⁻¹ schisantherin A were incubated with SK-HEP-1 cells, cell proliferation was analyzed by EdU DNA cell proliferation kit. Fluorescence probe method was used to assay the intracellular NO levels and NOS activity. Tube formation was observed using cell migration and a matrigel tube formation assay. Rat aortic ring assay was performed to observe the sprouting vessels from aortic ring. The fluorescence vessels, the number of functional blood vessels, and intersegmental vessel changes of transgenic zebrafish were also observed. Compared with control group, the proliferation of SK-HEP-1 cells induced by VEGF increased and and the level of NO and NOS activity induced; compared with model group, 2, 20 μmol•L⁻¹ schisantherin A and sorafenib inhibited the proliferation of SK-Hep-1 cells induced by VEGF, and reduced the level of NO and NOS activity. At the dosage of 20 μmol•L⁻¹, schisantherin A attenuated the migration and tube formation of SK-HEP-1 cells induced by VEGF, and also inhibition the formation of rat aortic rings and intersegmental vessel changes of transgenic zebrafish, and significantly reduce the number of vessels in zebrafish. Schisantherin A has potential effects on function of endothelial cell proliferation and angiogenesis.
ABSTRACT
Objective To establish a method for simultaneous determination of contents of schisandrin B, schisantherin A, lobetyolin and ruscogenin in Huangqi Shengmai Decoction with HPLC method. Methods The chromatographic column was an Agilent Eclipse C18 column (4.6 mm × 250 mm, 5 μm) that was maintained at a constant temperature of 30 ℃. The mobile phase was 0.1% acetic acid solution-acetonitrile, with gradient elution. The flow rate was 1.0 mL/min. The detection was at wavelength of 280 nm. Results The standard curves of schisandrin B, schisantherin A, lobetyolin and ruscogenin had good linearity in the ranges of 0.862 5-21.562 5μg (r 2=0.999 1), 0.737 5-18.437 5μg (r 2=0.999 4), 0.095 2-2.380 0 μg (r 2=0.999 6), and 0.810 0-20.250 0 μg (r 2=0.999 0), respectively. The average recoveries of the four components were 98.06%(RSD=1.64%), 99.61%(RSD=2.72%), 97.98%(RSD=1.45%), and 99.30%(RSD=1.25%), respectively. Conclusion This method is accurate, rapid and specific, achieving the stable results with high reproducibility.