| Morinda officinalis How., one of the "top four south authentic traditional Chinese medicines", is a commonly used Chinese herb distributed in the south of China, and its roots are widely used for the treatment of sexual impotence, spermatorrhea, irregular menstruation and female infertility. Previous phytochemical researches have revealed that anthraquinones, iridoids, saccharides are the main constituents of this specie.The pharmacology activities of Immunomodulation, anti-aging, anticancer, antidepressant, Fertility and oestrogenicity had been reported in modern pharmacology. TCM theory holds that Senescence is the kidney deficiency the first, then the spleen deficiency, and the qi and blood deficiency the last, therefore, to tonify kidney, invigorate spleen and benefit vital energy are the basic means of anti-aging. Morinda officinalis How.has a concrete manifestation of anti-aging effects on the its replenishing the kidney and strengthening the bones and muscles activities. To investigate the anti-aging active constituents in this plant, the study were performed.Although the chemical composition and pharmacological activities of M. Officinalis How.had been studied systematically, its anti-aging composition and mechanism is still unclear, in order to clarify the anti-aging active composition and the mechanism of this effects,the chemical composition of M. officinalis How. was studied, cell models of the chemical constituents were used for screening the different parts of M. officinalis extracted and its anti-aging effects were evaluated Synthetically.The studies provide the antiaging material foundation for the exploitation and application of M. officinalis and its preparations.This topic examined different M.officinalis extracts on the influence of mice swimming time and preliminary screened active site, the use of modern chromatographic techniques for the active site of its chemical constituents were isolated and purified by chemical and spectroscopic methods (UV, IR,1H-NMR,13C-NMR,2D-NMR, MS, etc.) to identify its structure.H2O2-induced cardiac cells in mice and human umbilical vein endothelial cell from aging model was used in this paper to screen compounds isolated from M. officinalis and evaluate their anti-aging effects, the most active single compounds were selected for anti-aging mechanism research and umbilical vein endothelial cells were employed for mechanism research. Cell viability was analyzed by using MTT assay, cell senescence was evaluated using galactosidase staining and telomerase activity detected by telomerase repeat amplification protocol(TRAP)-silver staining. The level of intracellular reactive oxygen species(ROS) was determined using fluorescent ROS detection kit and the level of klotho mRNA was detected by real time PCR.As a result,twelve compounds have been isolated, including six anthraquinones, two terpenoids, three saccharides and one coumarin. their structures were identified as: officinaliso(1), rubiadin-1-methyl ether(2), 1-hydroxy-6-hydroxymethylanthraquinone(3), anthraquinone-2-aldehyde(4), digiferruginol(5), 1-hydroxy-7-hydroxymethylanthraquinone(6),1,2-dihydroxy-3-methylanthraquinone (7), scopoletin(8), rotungenic acid(9), kestose(10),1F-fructofuranosylnystose(11),β-D-fructopyranosyl-(1→2)-β-D-fructofuranose (12). Those compounds were able to reduce the H2O2-induced cell senescence at different level, anti-aging activity of the twenty-four compounds from our team screened showed different levels of the compounds were able to reduce the H2O2-induced endothelial cell senescence,compound 2 manifested the strongest anti-aging effects,so compound 2 was selected for anti-aging mechanism research.MTT assay showed that with the concentrations of compound 2 (3-30μM) normal endothelial cells had no effect on survival,and 100μM compound 2 could significantly reduce the survival of endothelial cells by H2O2 (100μM) treatment with HUVECs,β-galactosidase activity and telomerase activity decreased significantly,different concentrations of compound 2 (3,10, and 30μM) or the positive control of resveratrol (30μM)could significantly inhibite the decreasing of telomerase activity in H2O2 (100μM)-induced; by H2O2 (100μM)-incubated treatment with HUVECs 48 h, the fluorescence intensity (reflecting the level of reactive oxygen species)was significantly enhanced, pretreatment with different concentrations of compound 2 (3,10, and 30 uM) or the positive control of resveratrol (30μM) could significantly inhibit the H2O2 (100μM)-induced elevation of intracellualr ROS level. By H2O2 (100μM) treatment with HUVECs 48 h, klotho mRNA expression did not change. Pretreatment with different concentrations of compound 2 (3,10, and 30μM) could significantly increase the expression of klotho mRNA, positive control of resveratrol (30μM) could upregulate the mRNA expression of klotho slightly.In this thesis,twelve compounds were obtained from M. Officinalis, compound 1 was a new compound, compound 12 was obtained from the genus of Morinda for the first time, compound 3,4,6,9 was isolated from this species for the first time,the NMR data of compound 3 is provided firstly in the literature. M.Officinalis's anti-aging activity screening effect is the comprehensive results of various components. The mechanism of anti-aging effect may be related to its effects of inhibiting oxidative function and upregulating the expression of klotho.
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