Study On The Material Basis Of CO₂-SFE From Angelica Sinensis To Protect Vascular Endothelial Cell

Angelica sinensis, a famous traditional Chinese medicine was extracted by CO2 supercritical fluid. Then, the chemical constituents of Angelica sinensis CO2-SFE were isolated by chromatographic methods and identified by spectroscopic methods. There were seven constituents being isolated and identified as following:(Z)-ligustilide, (Z)-butylidenephthalide, exo-Z,Z'-3a.7'a,7a.3'a-diligustilide, endo-Z,Z'-3a.7'a,7a.3'a-diligustilide, palmitic acid,3,9-dihydr oxy-ligustilide, Z,Z'-6.8',7.3'-diligustilide. Which should pay attention to was that endo-Z,Z'-3a.7'a,7a.3'a-diligustilide was found to be natural product and obtained from Angelica sinensis for the first time.To investigate the protective action of Angelica sinensis CO2-SFE and its compounds on the vascular endothelial cell, the endothelial cell strain of human umbilical vein (HUVEC) had been cultured in vitro. We established a model of endothelial cell oxidative damage induced by hydrogen peroxide (H2O2) and tested the influence of Angelica sinensis CO2-SFE and its compounds on the proliferation activity of HUVEC oxidative damage by the means of MTT assay. In model groups damaged by H2O2, some certain of injuries were observed. While the groups damaged by H2O2 (terminal concentration were 0.625,1.25,2.5,5.0,10.0mmol/L), the HUVEC showed obvious abnormalities. We selected the concentration of 1.25 mmol/L as our experimental concentration because the proliferation activity of HUVEC in this concentration was appropriate. Beside Angelica sinensis CO2-SFE can lessen proliferatory inhibition of HUVEC induced by H2O2, there were four compounds also can obviously protect HUVEC, they were ferulic acid, (Z)-ligustilide, endo-Z,Z'-3a.7'a,7a.3'a-diligustilide and 3,9-dihydroxy ligustilide.For study the possible anti-oxide mechanism, flow cytometry(FCM) and PI staining were used to analyzed cell cycle. Meanwhile, we used chemical methods to analysis the content of Lactate Dehydrogenase (LDH), the activity of super oxide dismutase (SOD), and the level of malanyldiadeyhde (MDA) and the content of nitric oxide (NO) in order to study the effect of Angelica sinensis CO2-SFE and endo-Z,Z'-3a.7'a,7a.3'a-diligustilide on HUVEC oxidative damage induced by H2O2.FCM showed that Angelica sinensis CO2-SFE and endo-Z,Z'-3a.7'a,7a.3'a-diligustilide can increase cells at S phase, decrease cells at G2 phase, inhibit the influence of H2O2 which decrease cells at S phase, increase cells at G2 phase. Angelica sinensis CO2-SFE and endo-Z,Z'-3a.7'a,7a.3'a-diligustilide also can decrease the level of MDA and LDH, promote the activity of SOD, increase the excretion of NO.According to these studies, they can indicate that Angelica sinensis CO2-SFE and endo-Z,Z-3a.7'a,7a.3'a-diligustilide has the protective action on oxidative damage of HUVEC induced by H2O2.This study provided new theoretical basis to the prevention and treatment of atherosclerosis.