Screening The Anti-aging Active Components From The Fruit Of Phellinus Baumii And Study On The Mechanism Of Active Compounds

Phellinus baumii, which is always being named Sang huang Gu, Sangchen,Sang er in Chinese, is an edible and medicinal fungi belonging to Basidiomyeotina. Phellinus baumii has the laudatory title of gold forest. Mainly chemical compounds such as flavonoids, triterpene and sugar were included. In Traditional Chinese medicine, Phellinus baumii was widely used to treat metrorrhagia, amenorrhea, diarrhea pixu. More recent researches have shown that Phellinus baumii has possessed immunomodulatory properties, anti-oxidant activity, antitumor and many other pharmacological effects. Modern research about chemical composition and pharmacological action were studied systematically, but anti-aging effect mechanism is still unclear. In this paper, repairing nerve cell capacity and the ability to remove free radicals were used to Screen the anti-aging active compounds from the fruit of Phellinus baumii in order to explore the mechanism of these compounds. This thesis is divided into four parts, the main findings are as follows:1. Compare different biological activity from different the extraction phase in order to obtain higher antioxidant activity phase.Petroleum ether, chloroform, ethyl acetate, n-butyl alcohol and were obtained after ethanol extracts of Phellinus baumii fruit bodies (1.5L,241.84g) were extracted by four different organic solvent in the same volume. Quality of extraction of extract was obtained:6.29g,12.02g,19.67g and19.37g inorder. Both repairing damaged nerve cells in vitro and the ability to remove free radicals were considering to find good anti-aging activity extraction phase. The experimental results show that the four extraction phase there was some difference of anti-aging ability. The ethyl acetate and n-butanol phase activity were better than the other two phase extraction. Ethyl acetate and n-butyl alcohol phase extraction existed ignificant difference about the aiility of repairing nerve cells (P<0.05). The IC50value of ethyl acetate extraction in scavenging superoxide, H2O2and DPPH were589.17μg/mL,5.60μg/mL,209.00μg/mL,whileThe IC50value of n-butyl alcohol extraction were483.12μg/mL,0.42μg/mL,56.22μg/mL.2, Purification and structural analysis of the anti-aging components from ethyl acetate extractionThe ethyl acetate phase which showed higher anti-aging t capacity were selected in the further purification experiment. The silica gel, the Sephadex LH-20and other methodse were used to purify the samples.11compounds were obtained: Ergosta-7,22-dien-3β-ol, ergosta-6,2-dien-3p,5a,8a-triol, Ganoderiol B, Ganoderic acid DM, inoscavin A and Ergosta-7,22-dien-3β-yl pentadecanoate.3、A single compound anti-aging activity of studyThe anti-aging activity of purified compound was determined by the ablity of repairing damaged nerve cells and scavenging free radicals.There was highly significant difference (P<0.05) in the ablity of repairing damaged nerve cells between compound1, compound10and compound11at the concentration of50μg/mL. Meanwhile,there was also significant difference (P <0.05) between compound5, compound8,compound10and compound11at100μg/mL. P<0.05indicates the compounds showed some certain effects of repairing damaged nerve cells.Four single compounds showed relatively high superoxide anion scavenging activity and IC50value of compound5, compound8, compound10and compound11was23.69μg/mL,131.38μg/mL,67.89μg/mL,52.69μg/mL respectively. Other compounds showed lower superoxide anion scavenging activity. The four compounds also showed high DPPH radical scavenging activity with IC50value was1.22μg/mL,0.83μg/mL,3.00μg/mL,1.47μg/mL respectively. Finally,we screen compound10and compound11as anti-aging high anti-aging compounds by in vitro repairing damaged nerve cells and free radicals scavenging experiments.4、Study on the Anti-aging mechanism of active compoundsAging may lead to a series of diseases, such as Alzheimer’s disease. This thesis uesd dementia as research background, discussed how the single compounds might actually affected expression of several key enzymes in this disease. The results showed that comparing the other samples under the same concentration, Compound10and11inhibition rate of AchE was generally higher. Western blot results also showed that Compound10also can inhibit the expression of BACE1and HDAC family protein in senescent cells.This subject offered a new way to slow the aging and also provided scientific basis for exploitation and utilization of Phellinus baumii.