Preliminary Study On Chemical Constituents In Roots Of Three-year-old Glehnia Littoralis

Glehnia littoralis Fr. Schmidt ex Miq. is a perennial herb, which belongs to Glehnia F. Schmidt (Umbelliferae) in plant taxonomy. The dried roots of Glehnia littoralis, known as a kind of traditional Chinese medicine, were used for nourishing yin to clear away the lung-heat and tonifying stomach and promoting fluid. In this research, the roots of three-year-old Glehnia littoralis, which were dried in the shade without being removed the peel, were chosen as materials, and used in this preliminary study.Constituents in roots of three-year-old Glehnia littoralis were extracted with 95% ethanol, then the extract were suspended in water. Petroleumether, ethyl acetate and n-butanol were successively used as solvents to extract constituents from the suspension. Silica gel column chromatography, Sephadex, silica gel thin-layer chromatography, prepared silica gel thin-layer chromatography, recrystallization and reverse phase silica gel C-18 were used in isolatation of chemical constituents. Nuclear magnetic resonance were used in elucidation of the chemical structures of compounds isolated from roots of three-year-old Glehnia littoralis. And seven compounds had been identified, which were bergapten, stigmasterol, nonadecanoic acid, tetracosanoic acid, cerevisterol.5-methoxyfurfural and scopoletin. Among them, nonadecanoic acid, tetracosanoic acid, cerevisterol,5-methoxyfurfural were isolated from the roots of Glehnia littoralis for the first time.High performance liquid chromatography was used in this study to determine and compare the contents of stigmasterol separately in one-year-old, two-year-old, three-year-old roots of Glehnia littoralis and root of one-year-old Glehnia littoralis without peel. And the results showed that the content of stigmasterol in root of one-year-old Glehnia littoralis without peel was 0.3775mg·g-1 and was the highest. And the content of stigmasterol in root of one-year-old Glehnia littoralis was 0.2796mg·g-1, and in root of two-year-old Glehnia littoralis the content was 0.0975 mg·g-1, which declined significantly compared to the content in the one-year-old one. Root of three-year-old Glehnia littoralis had the lowest content of stigmasterol and that was 0.0691 mg·g-1. The content of stigmasterol in one-year-old root of Glehnia littoralis without peel was 1.35 times of the content of stigmasterol in one-year-old root of Glehnia littoralis with peel,3.87 times of the content of stigmasterol in the two-year-old Glehnia littoralis and 5.46 times of the three-year-old Glehnia littoralis. The content of stigmasterol had negative correlation with the number of grown years of Glehnia littoralis. Therefore, in production aimed at stigmasterol, one-year-old roots of Glehnia littoralis should be chosen as raw material and it would be better these roots were blanched and removed peels.In vitro antitumor activities on liver cancer cell line HEP-G2 and human stomach cancer cell line SGC-7901 of bergapten or stigmasterol were examined using MTT method separately. And the results showed that bergapten had inhibitory effect to the growth of HEP-G2 at all five concentration levels from 0.01 to 100.00mg·L-1. And the inhibitory rate of the bergapten solution with concentration of 100.00 mg·L-1 was 95.0%, which was significantly higher than the inhibitory rates of bergapten solutions with other concentrations. Only bergapten solution with concentration of 100.00 mg·L-1 had inhibitory effect to SGC-7901 and the inhibitory rate was 79.6%, while bergapten solutions with other concentrations had no inhibitory effect to this cancer cell line. Stigmasterol had inhibitory effect to the growth of HEP-G2 at all five concentration levels from 0.01 to 100.00mg·L-1. And the inhibitory rate of the stigmasterol solution with concentration of 100.00 mg·L-1 was 93.0%, which was significantly higher than the inhibitory rates of stigmasterol solutions with other concentrations. To SGC-7901, stigmasterol had no inhibirory effect at any concentration level. For both bergapten and stigmasterol, inhibitory effect was dose-independent. In other words, inhibitory effect had positive correlation with concentration.