Studies On Constituents And Biological Activities Of Diospyros Kaki L.f.

"ShiYe"is the fresh or dry leaf of Diospyros kaki L. f. (Ebenaceae). It is employed in the treatment of hypertesion, heart disease, the internal haemorrhage induced by gastric ulcer, and pulmonary tuberculosis, the functional bleeding of ulterus and so on. Its chemical constituents is very complex, mainly including flavonoids, triterpernoids, phenols, etc.43 compounds were isolated from the leaves of Diospyros kaki with porous resin (D101), silical gel, sephadex LH-20, polyamide, HPLC, PTLC etc. Among them, 34 compounds were identified on the basis of chemical and spectral methods as: 4, 6-Dihydroxy-2-O-β-D-glucopyranosyl-l, l'-diphenyl-one (1) and 4', 5-Dimethoxy-3-O-β-D-glucopyranosyl-4-hydroxy-biphenyl (2), 4, 4'-Dihydroxy-a-truxillic acid (3), Tatarine C (4), Linarionoside A (5), Linarionoside B (6), Annulatin (7), Isorhamnetin-3-O-β-D-glucopyranoside (8), Kampferol-3-O-β-D-galactopyranoside (Trifolin) (9), Kampferol-3-O-β-D-glucopyranoside (Astragalin) (10), Kampferol (11), Quercetin-3-O-β-D-galactopyranoside (Hyperin) (12), Quercetin-3-O-β-D-glucopyranoside (Isoquercetrin) (13), Rutin (14), Quercetin (15), Vitexin (16), 2"-O-Rhamnosylvitexin (17), a-Amyrin (18), p-Amyrin (19), Lupeol (20), Uvaol (21), Oleanolic acid (22), Ursolic acid (23), Betulinic acid (24), Pomolic acid (25), 19a, 24-Dihydroxy ursolic acid (26), Barbinervic acid (27), Rosamultin (28), Daucosterol (29), p-Sitosterol (30), 3, 4'-Dihydroxy benzoic acid (31), Benzoic acid (32), p-Hydroxy benzoic acid (33), Succinic acid (34). Among them, 1 and 2 were new compounds, named as Kakispyrone and Kakispyrol; 3, 4, 5, 6, 7, 8, 16, 17, 26, 27, 28 were isolated from this genus for the first time; 3, 4, 5, 6, 7, 8, 16, 17, 18, 19, 20, 25 , 26 , 27 , 28 were isolated from this plant for the first time.A review on the development of research about chemical constituents and pharmacological activities of Diospyros L. plants was provided. The regulations of C-glycosylflavones structure elucidation on the basis of UV NMR MS CD spectra and chemical method, the components and activation mechanisms of human NADPHOxidase, the relation of free radical and disease were summarized respectively.Antioxidation bioactivity of five triterpenoid compounds (18, 21, 23, 25, 26) and five flavonoid compounds (8, 9, 10, 12, 14) isolated from the leaves of Diospyros kaki L. were tested. On the basis of experiment data, the link of structure and activity was discussed. We found that these compounds suppressed the superoxide generation in human neutrophils induced by N-formyl-methionyl-leucyl-phenylalanine (fMLP) in a concentration-dependent manner, especially compound 25 showed the strongest effect. Compound 8, 9, 12, 14, 23, 25 and 26 suppressed the superoxide generation induced by phorbol 12-myristate 13 -acetate (PMA) and compound 25 showed the strongest effect, but compound 10, 18 and 21 had no effect. Arachidonic acid (AA) induced superoxide generation was suppressed by compound 9, 10, 12 and 14; Compound 8, 18, 21, 23, 25 and 26 weakly enhanced AA-induced superoxide generation in low concentration but suppressed it in high concentration.Additionally, we found compound 8, 9, 14, 23, 25 and 26 inhibited fMLP-induced tyrosyl phosphorylation of 45 kDa protein in human neutrophils, which explained the suppression mechanism of the superoxide generation.It was found that the five flavonoid compounds showed no hemolytic effect even at 100 M and 500 M, which was much higher than the concentrations sufficient for the suppression of superoxide generation.