Extracts And Biological Activities Of Sporodem-broken Spores Of Ganoderma Lucidum

Ganoderma lucidum spores (GS) shot out from mature fruit bodies are often covered by two rigid sporodems which have limited their values. Recently, people began to explore their components and bioactivities with the development of sporodem-broken and gathering technology. However, the mechanism of some activities still haven’t been clearly expounded. In this study, we investigated the activities of sporodem-broken spores and their by-products aiming to make the best use of these natural products and lay theoretical foundation for their further development.We firstly analyzed the extracts of GS. Ethanol extract of GS (EGS) was prepared by 80%ethanol and analyzed. The results showed that the yield was about 6.09±0.42% and it contained 13.56±1.23%triterpene acids,14.10±0.92% polyphenols,13.92±0.73% saccharides and 2.09±0.13% proteins. The residue was re-extracted with water to separate polysaccharides, namely the aqueous part of GS (AGS) and orthogonal test L9(34) was used to determine the best extraction conditions of AGS. The results showed that the optimum conditions were 1:20 solid/liquid ratio,100℃,3 times and 2h per time, under which the yield was 3.21±0.21% and the purity reached to 67.47%.To investigate the activities of GS, free radical scavenging ability of AGS and EGS was measured then. The results indicated that the T-AOC, DPPH· and ·OH scavenging rate of AGS at 6mg/mL were 25.04 U/mL,64.32% and 79.47%, respectively; while those of EGS at 1mg/mL were 23.67 U/mL,82.22% and 65.88%, respectively. Furthermore, the results of anti-lipid peroxidation studies showed that both AGS and EGS significantly suppressed erythrocyte hemolysis self-motivated or H2O2-induced in a dose-dependent manner, as well as decreased the level of MDA induced by H2O2 in mouse liver in vitro, indicating that GS can protect erythrocytes and liver from oxidative damage. These results suggested that GS was a potential vector of natural antioxidants.Antitumor activity of AGS and EGS was examined by SRB method on three lung cancer cell lines. Results showed that it was EGS but not AGS that significantly inhibited cell growth and survival in a dose-dependent manner, so we took EGS for further studies. In these studies, EGS arrested cell cycle at G2/M phase by down-regulation the expression of CyclinB1 and p-Cdc2 as well as directly induced cancer cell apoptosis through suppression the level of Bcl-2 and Bcl-xl. Additionally, EGS inhibited lung cancer cell migration by Transwell assay. Both GS and EGS significantly suppressed tumor volume without body cytotoxicity in a human lung cancer A549 cell xenograft mice model, and decreased the level of Ki67. Further studies showed that GS decreased the level of p-Akt, p-mTOR, p-S6 and p-4EBP1 without down-regulating the level of total proteins, indicating that EGS targeted to Akt/mTOR signaling pathway. Taking together, these results suggested that GS blocked cell cycle and induced apoptosis via Akt/mTOR signaling pathway, leading to the inhibition of lung cancer cell growth and survival in vivo and in vitro.The by-products of GS were also studied to screen out the most efficient exact. Ethanol soluble extract was prepared and follow by fractional extraction to get the other four parts, namely petroleum ether, chloroform, ethyl acetate and n-butanol. The most efficient extract was ethyl acetate part screened by T-AOC and free radical scavenging assays. Then the ethyl acetate part was taken a further study on anti-lipid peroxidation ability. The results indicated that each extract had significant antioxidant ability especially for the ethyl acetate extract.