Study On The Anti-aging Constituents And Mechanism Of Damnacanth Us Officinarum Huang.

Damnacanthus officinarum Huang (DOH; Ensi Ba-jitian) is a plant of Rubiaceae family, which has been extensively used as a valuable Yang-tonic agent in Hubei, Hunan and Sichuan provinces for medicinal purposes as an alternative of M. officinalis root, which is one of four famous "southern medicines" in China. As a Yang-tonic agent, DOH root has been used to nourish "kidney" and strengthen "bone and muscle", which are important indices for impotence and infertility in traditional Chinese medicine. To evaluate the neuroprotective and anti-aging activity of extracts in Caenorhabditis elegans from the roots and leaves of Damnacanthus officinarum Huang, and to provide the pharmacological basis in traditional medicine, we systematically studied the chemical constituents and the medicinal effect of DOH.It has been shown that the ethanol, n-butanol and aqueous extracts in the roots and leaves possessed significantly neuroprotective effect both in chemosensory behavior test. In ASH neuron survival model, the ER, BR, AR and EL extracts were able to protect ASH neuron from polyQ-mediated death. Results of neuroprotective and chemosensory behavior test were consistent, among which the n-butanol extracts showed best efficacy. Moreover, the neuroprotective and lifespan-extention activity effects of root extracts are superior to leave extracts, supporting the traditional application of above-ground parts of DOH in treating various diseases associated with brain disorders and anti-aging.Four candidate extracts, possessing excellent neuroprotective activity, extend lifespan in C. elegans. Compared with the lower dose group (800ug/ml), the higher dose group (1200ug/ml) showed better significant effect of extending lifespan in C. elegans. The root of the n-butanol of the higher dose group (1200ug/mL) enabled nematodes to extend the average life expectancy of 30.3%.The pathophysiological process of a variety of neurodegenerative diseases associated with ROS, especially the aging brain tissue vulnerable to ROS oxidative damage. Treatment with BR and AR(1200ug/ml) extracts increased the survival rate of the worms exposed to paraquat. It should be speculated that the anti-aging mechanisms of DOH would be closely related with neuronal protection, in part related to antioxidant and free radical scavenging pathways in Caenorhabditis elegans.Our chemical constituents study of DOH afforded 17 compounds. They were isolated and purified by chemical and spectroscopic methods (NMR,MS,IR,UV etc.) to identify their structures. They wereβ-sitosteol(DOH 1), Lignoceric acid (DOH 2), 1-hydroxy-2-hydroxy-methylanthraquinone (DOH 3), Oleanolic acid acetate (DOH 4), Alizarin dimethyl ether (DOH 5), Morindaparvin A (DOH 6), Daucosterol (DOH 7),2-hydroxy-l-methoxyanthraquinone (DOH 8),2,5-dihydroxy-1-methoxy-anthraquinone (DOH 9), n-butyl-β-D-fructopyranoside (DOH 10), Monotropein (DOH 11), (1S,5S,9S)-Δ3(4),7(8)-10-dihydroxymethyl-4-iridoid acid-1-O-β-D-glucose (DOH 12), (digiferruginol-l-methylether-11-O-β-gentiobioside, (DOH 13), digiferruginol-11-O-β-primeveroside (DOH 14), stearic acid (DOH 15), a-D-Glucose (DOH 16), (3-D-Glucose (DOH 17), Including seven anthraquinones and anthraquinone glycosides, two iridoids, a triterpenoid. In this thesis, DOH 12 was a new compound, DOH 2,4,5,7,9,10,11,13,14,15,16,17 were obtained from Damnacanthus officinarum Huang for the first time, and were discovered from the Damnacanthus species for the first time. This subject further clarified the material basis of Damnacanthus officinarum Huang, this may provide valuable evidence for its traditional use in treating various diseases or dietary therapy in China.