ABSTRACT
Objective: To study the suspension culture of tuber and its alkaloid content based on the stimulation of salicylic acid. Method: The tubers of Pinelliae Rhizoma in suspension tube were treated with different concentrations of exogenous salicylic acid to analyze the growth status. The content of alkaloids in tuber was detected by HPLC. Test conditions:chromatographic column for Agilent Eclipse plus C18 column (4.6 mm×250 mm,5 μm),mobile phase of acetonitrile water(4:96),the column temperature was maintained at 35℃,detection wavelength for inosine 250 nm,guanosine 260 nm,volume flow rate 1.0 mL·min-1. Result: The results showed that the exogenous salicylic acid had a certain effect on the growth of suspension tuber of Pinelliae Rhizoma. When the salicylic acid concentration was 150 μmol·L-1,the culture lasted for 25 days and the fresh weight reached the maximum value of 7.483 8 g. It also accumulates a certain amount of alkaloids. The linear range of guanosine was 0.03-0.45 μg (R2=0.999 6). After 10-days cultivatation in the salicylic acid concentration of 50 μmol·L-1,guanosine content of Pinelliae Rhizoma tubers reached a maximum of 1.353 3 mg·g-1. The linear range of inosine 0.003-0.045 μg (R2=0.999 5). When the salicylic acid concentration was 200 μmol·L-1,cultured for 30 days,the content of inosine in Pinelliae Rhizoma tubers reached the maximum value of 0.149 8 mg·g-1. Conclusion: The results of this experiment provide a reference for the study of tissue culture and rapid propagation of Pinelliae Rhizoma tubers and regulation of alkaloids,which are of great significance for the development of Pinelliae Rhizoma industry.
ABSTRACT
Objective: To study the effect of menthol on proliferation, migration and expressions of interleukin-8(IL-8), C-X-C motif chemokine-12(CXCL-12) and vascular endothelial growth factor(VEGF) in hepatoma HepG2 cells in vitro, in order to elucidate the anti-liver cancer activity of menthol and relevant mechanisms. Method: Different concentrations of menthol (25, 50, 100, 200 μmol·L-1) and blank group were applied to Hepatoma HepG2 cells. Cell counting kit-8(CCK-8) and EDU were used to detect the proliferation effect of menthol on HepG2 cell. Transwell experiment was used to detect the migration effect of menthol on HepG2 cell. Real-time fluorescent quantitative polymerase chain reaction technique was used to detect the inflammatory chemokines IL-8 and CXCL-12 mRNA expression levels in HepG2 cell. Western blot was used to detect the expression level of VEGF in HepG2 cells treated with menthol. Result: Compare with the blank group, menthol (25, 50, 100, 200 μmol·L-1) significantly inhibited proliferation and migration of HepG2 cells in vitro. When the concentration of menthol was 100 μmol·L-1 microns, the inhibitory effect was significant (P-1) significantly inhibited the expression levels of IL-8, CXCL-12 mRNA and VEGF protein in HepG2 cells (PConclusion: Menthol has an inhibitory effect on the proliferation and migration of hepatoma HepG2 cells in vitro, and the potential anti-liver cancer mechanism might be related to the inhibition of IL-8, CXCL-12 and VEGF expressions in the cells. This conclusion provides the experimental basis for elucidating the effect of menthol against liver cancer.