Establishment of quantitative analysis of multi-components by single marker for content determination of 10 ganoderic acids in Ganoderma lucidum / 中国药房

OBJECTIVE To establish a quantitative analysis of multi-components by single marker （QAMS） method for simultaneous determination of 10 ganoderic acids in Ganoderma lucidum. METHODS Using ganoderic acid A as internal reference， high-performance liquid chromatography （HPLC） method was adopted to calculate relative correction factors of the other 9 components， such as ganoderic acid B， ganoderic acid C2， ganoderic acid D， ganoderic acid F， ganoderic acid H， ganoderenic acid A， ganoderenic acid B， ganoderenic acid C， ganoderenic acid D； the contents of above ganoderic acids were calculated with relative correction factors， and compared with the results of external standard method. RESULTS The linear relationship of ganoderic acid A， ganoderic acid B， ganoderic acid C2， ganoderic acid D， ganoderic acid F， ganoderic acid H， ganoderenic acid A， ganoderenic acid B， ganoderenic acid C and ganoderenic acid D were 0.032-3.996， 0.040-4.971， 0.037-4.568， 0.028-3.558， 0.033-4.177， 0.044-5.440， 0.032-3.944， 0.040-4.994， 0.045-5.593 and 0.035-4.342 mg/mL （all R 2≥0.999 2）， respectively. RSDs of precision， stability （24 h） and reproducibility tests were all lower than 2%. Their average recovery rates were 99.43%， 100.25%， 98.50%， 99.88%， 100.59%， 99.64%， 98.50%， 99.40%， 99.64% and 99.76%， respectively （RSD＜2%， n＝6）. Relative correction factors of ganoderic acid B， ganoderic acid C2， ganoderic acid D， ganoderic acid F， ganoderic acid H， ganoderenic acid A， ganoderenic acid B， ganoderenic acid C and ganoderenic acid D were 1.788 5， 1.288 2， 1.126 4， 1.698 5， 0.885 4， 5.468 1， 4.210 9， 5.780 8， 4.290 3， respectively. Relative errors between the content obtained by QAMS method and external standard method for G. lucidum from different origins were within ±12%. CONCLUSIONS It is feasible that the contents of 10 ganoderic acids are determined simultaneously by QAMS method， using ganoderic acid A as internal reference. This method shows good precision and reproducibility and can be used for the quality control of G. lucidum.

Germplasm resources and secondary metabolism regulation in Reishi mushroom (Ganoderma lucidum) / 中草药·英文版

Ganoderma lucidum is a valuable medical macrofungus with a myriad of diverse secondary metabolites, in which triterpenoids are the major constituents. This paper introduced the germplasm resources of genus Ganoderma from textual research, its distribution and identification at the molecular level. Also we overviewed G. lucidum in the components, the biological activities and biosynthetic pathways of ganoderic acid, aiming to provide scientific evidence for the development and utilization of G. lucidum germplasm resources and the biosynthesis of ganoderic acid.

Research progress on pharmacological activities of triterpenes from Ganoderma lucidum / 中国药理学通报

Ganoderma lucidum is a valuable traditional Chinese medicine with dual use of medicine and food in China. Its chemical components mainly include triterpenoids, polysaccharides, organic acids, alkaloids, amino acids and so forth. The triterpenoids mainly include ganoderma acid and ganoderma alcohol. This paper has summarized the pharmacological activities of Ganoderma lucidum triterpenes in anti-tumor, anti-inflammatory, liver and kidney protection, immune regulation, blood lipid and blood glucose lowering, and antimicrobial activities in recent years, in order to provide reference basis for the core efficacy e-valuation of high-quality Ganoderma lucidum. It also provides scientific evidence for the application of Ganoderma lucidum in health food and clinical medicine.

Ganoderic acid A protects lens epithelial cells from UVB irradiation and delays lens opacity / 中国天然药物

A contributory role of oxidative stress and protection by antioxidant nutrients have been suspected in cataract formation. Ganoderic acid A (GAA), an effective lanostane triterpene, is widely reported as an antioxidant. The aim of this study is to investigate the potential effects of GAA on cataract formation. After lens epithelial cells (LECs) were exposed to UVB radiation for different periods, cell viability, apoptosis-related protein levels, malondialdehyde (MDA) and superoxide dismutase (SOD) activities were monitored. We found that cell viability, the Bcl-2/Bax ratio and SOD activity were increased, while Cleaved caspase-3 levels and MDA activity were decreased compared with those in UVB-impaired LECs after GAA treated. Furthermore, GAA activated PI3K/AKT in UVB-impaired LECs and effectively delayed the occurrence of lens opacity in vitro. In conclusion, these findings demonstrated that GAA exhibited protective functions in SRA01/04 cells and rat lenses against UVB-evoked impairment through elevating cell viability and antioxidant activity, inhibiting cell apoptosis, activating the PI3K/AKT pathway and delaying lens opacity.

Study on the Effect and Mechanism of Ganoderic Acid C 2 on Lipid Metabolism of Hepatocytes by Regulating S6K/SREBPs Signaling Pathway / 中国药房

OBJECTIVE：To stu dy in vitro lipid-lowering effect of ganoderic acid C 2（GAC2），and to investigate its potential mechanism on the basis of S 6K/SREBPs signaling pathway. METHODS ：Using human liver cells HL- 7702 as objects ，MTT assay was used to test relative cell viability after treated with low ，medium and high doses （5，10，20 μmol/L，hereinafter）of GAC 2. Using lovastatin as positive control ，ELISA method was used to detect the contents of TC and TG in cells after treated with low ， medium and high doses of GAC 2. Nile red staining was used to observe the accumulation of lipids in cells. After transfected SREBPs report gene plasmid ，using 25-HC as positive control ，relative viability of SREBPs luciferase in cells were determined by luciferase assay after treated with low ，medium and high doses of GAC 2. Using 25-HC as positive control ，real-time fluorescent quantitative PCR was used to measure the mRNA expression of SREBPs and their downstream genes in cells after treated with medium and high doses of GAC 2. Using SREBPs inhibitor （25-HC）and S 6K inhibitor （rapamycin）as control ，Western blotting assay was adopted to determine the expression of SREBP- 1 and SREBP- 2（in the case of n-SREBPs ），relative expression ratio of phosphorylated S 6K to S 6K（p-S6K/S6K ratio ）. AutoDock 4.0 and other softwares were used for molecular docking of S 6K and GAC2. RESULTS ：There was no significant effect of low ， 0.05）. Compared with blank control group ，the content of TC qq.com in lovastatin group and GAC 2 high-dose group as well as thecontent of TG in lovastatin group ， GAC2 medium- and 床应用。电话：0371-65962746。E-mail：whui3697@126.com high-dose groups were decreased significantly （P＜0.05 or P＜ 0.01）；the number of lipid droplets in the cells of all medication groups decreased. Compared with blank control group ，relative viability of SREBPs luciferase in 25-HC group ，GAC2 low-，medium- and high-dose groups were decreased significantly ；mRNA expression of HMGCS1，MVK，SCD，HMGCR gene in 25-HC group and GAC 2 medium-，high-dose groups ，mRNA expression of DHCR7 gene in 25-HC group ，mRNA expression of SREBP-2 gene in GAC- 2 high-dose group as well as mRNA expression of DHCR24 and MSMO2 gene in 25-HC group and GAC 2 high-dose group were all decreased significantly ；relative protein expression of n-SREBP- 1 in 25-HC group ，GAC2 low-，medium- and high-dose groups ，relative protein expression of n-SREBP- 2 in 25-HC group and GAC 2 high-dose group as well as p-S 6K/S6K ratio in rapamycin group and GAC 2 groups were decreased significantly （P＜0.05 or P＜0.01）. The molecular docking results showed that GAC 2 could bound to amino acid residues Arg 335，Arg330 and Ala332 of S 6K via hydrogen bond. CONCLUSIONS ：GAC2 can reduce the lipid level of HL- 7702 cells，which may be associated with inhibiting the expression of S 6K/SREBPs signaling pathway.

A new meroterpenoid from Ganoderma resinaceum / 中草药

Objective: For the purpose of finding new agents, the chemical study on Ganoderma resinaceum was carried out. Methods: The chemical constituents from the dried fruiting bodies of G. resinaceum were isolated by column chromatographic methods of silica gel, MCI-Gel resin, and high performance liquid chromatography. The structures were elucidated by spectroscopic methods, including extensive 1D and 2D NMR techniques. Results: Four compounds including one new meroterpenoid were isolated from this fungus. Their structures were identified as ganoresinains F (1), 3β,7β,15β-trihydroxy-11,23-dioxo-lanost-8,16-dien-26-oic acid (2), ganoderic acid XL2 (3) and 2,3-dihydro-4(1H)-quinolone (4). Conclusion: Compound 1 is a new compound and named as ganoresinains F (1).

Pharmacokinetics of ganoderic acids / 中国中药杂志

Ganoderic acid(GA) is one of main bioactive components produced by Ganoderma lucidum,which a traditional Chinese herbal medicine and a kind of tracyclic triterpene lanosterol derivatives with highly oxidized structure. It has a variety of important pharmacological activities,such as anticancer,immunoregulation,anti-oxidation,anti-diabetes and anti-HIV. At present,the studies of GA mainly focus on biosynthesis,fermentation control,isolation and purification,structure identification and pharmacological effects.However,there are a fewer pharmacokinetic studies of GA,although it is closely related to the clinical application. Recent studies have shown that GA can be absorbed rapidly by gastrointestinal tract and distributed in various tissues and organs after oral intake. GA is metabolized by liver at phase Ⅰ and phase Ⅱ,and then mainly excreted by bile. In this paper,the pharmacokinetic characteristics of GA and its absorption,distribution,metabolism and excretion(ADME) will be systematically summarized,in order to provide scientific basis for the application and development studies of Ganoderma triterpenoid drugs and their rational clinical use.

Ganoderic acid A from Ganoderma lucidumretard ADPKD renal cyst development via down-regulating Ras/MAPK signaling pathway / 中国药理学与毒理学杂志

OBJECTIVE Autosomal dominant polycystic kidney disease (ADPKD) is a common-monogenetic disease characterized by progressive development of renal cysts. Thereis still further need for effective therapy.Based on our precious study that Ganoderma triterpenes(GT),which is the major secondary metabolites of Ganoderma lucidum,is able to attenuate renal cyst development.The aim of this study was to investigate the effect of a monomer,Ganoderic acid A(GA-A)that was purified from the GT,which has been reported to exhibit antinociceptive,antioxidative,hepatoproctive and anti-cancer activities,to have a potent anti-cyst effect in ADPKD. METHODS We first evaluated the potential cytotoxicity of GA-A on MDCK cells using a CCK-8 assay.Then we used MDCK cyst model,cultivated MDCK cells in vitro to form fluid-filled cysts surrounded by monolayer cells.MDCK cells were co-incu-bated with 10 μmol·L-1FSK with or without GA-A(25 μg·mL-1)and equal concentration GT as positive control from day 0 to day 6 to investigate the inhibitory effect of GA-A on cyst formation.And to further investigate the inhibitory effect of GA-A on cyst enlargement, MDCK cysts were treated with different concentration of GA-A(6.25,25 and 100 μg·mL-1)from day 5 to day 12.Next we used an embryonic kidney cyst model, wile-type mice kidneys were taken out on embryonic day 13.5 to form renal cysts stimulated with 8-Br-cAMP to prove the renal cyst inhibition at organ level.Meanwhile,we explored the possible mechanisms underlying GA-A inhibition on renal cyst development using MDCK cells treated with 10 μmol·L-1FSK co-incubated with GA-A(25 μg·mL-1)and equal concentration GT.Several key components of Ras/MAPK pathway was evaluated by Western blot,the protein expression of H-ras,B-raf, p-ERK, Egr-1 and c-fos was evaluated. RESULTS MDCK cell viability was not affected by GA-A that were used ofincreasing concentrations up to 200 μg·mL-1. GA-A had no significant influence on cyst formation,but inhibited cyst enlargement dose-dependently and the inhibitory effect is significantly better than that of the same concentration of GT which proved that GA-A may be an effective monomer from GT.And after washing out GA-A on day 8,MDCK cysts regrew to a large size,suggesting that the inhibitory effect of GA-A on MDCK cyst enlargement was reversible. GA-A inhibited embryonic kidney cyst development significantly in a dose-dependent and reversible manner proving GA-A cyst inhibition at organ level,which is also more effective than equal concentration GT.Treatment of MDCK cells with FSK caused a significant elevation of H-ras,B-raf,p-ERK,Egr-1 and c-fos signaling molecules,while treatment with GA-A reduced the level of H-ras,B-raf,p-ERK,Egr-1 and c-fos expression.GA-A down-regulated Ras/MAPK signaling pathway could contribute to its inhibitory effect on cyst development. CONCLUSION Ganoderic acid A from Ganoderma lucidum retard ADPKD renal cyst development via down-regulating Ras/MAPK signaling pathway.

Preparation of ganoderic acid T tablets and pharmacokinetics in rabbits / 中国药学杂志

OBJECTIVE: To prepare the ganoderic acid T (GA-T) tablets and evaluate the pharmacokinetics and bioavailability in rabbits. METHODS: GA-T tablets were prepared by direct powder compression. After investigating the type and amount of excipients by single-factor method, the formulation of GA-T tablets was optimized and prepared by direct powder compression. An HPLC method was used for the determination of plasma concentration in rabbits after administration of GA-T. The main pharmacokinetic parameters were analyzed by pharmacokinetic software kinetica 5.0. The pharmacokinetic characteristics of GA-T tablets were studied. RESULTS: In optimization of formulation, microcrystalline cellulose and spray-dried lactose (1:1) were selected as bulking agent. In addition, 1% of the silica powder and 0.5% magnesium stearate were used as glidant and lubricant, respectively. The prepared tablets were consistent with the requirements of the 2010 edition of Chinese Pharmacopoeia standards, including appearance, hardness, disaggregation and dissolution of the tablets. The absolute bioavailability of oral GA-T tablets was 33.03%. CONCLUSION: The direct powder compression is simple and reliable, and the bioavailability of oral GA-T tablets in rabbits is well. This article provided experimental data for preclinical study of GA-T.

Determination of ganoderic acid B in different parts of Ganoderma lucidum / 中草药

To explore the distribution of the bitter principle ganoderic acid B in different parts of Ganoderma lucidum (Leyss. ex Fr.) Karst.. Methods　Ganoderic acid B in epidermis, phellem, stipe, spore and pileus were determined by RP-HPLC. Results　Ganoderic acid B was found to be mainly concentrated in the entire surface of the basidiocarp, other parts contained much less amount. Conclusion　Result of the study may provide references for the search of such active principle from this fungus.

Chemical constituents from fruiting bodies of Ganoderma tsugae (Ⅱ) / 中草药

Objective To study the chemical constituents from the fruiting bodies of Ganoderma tsugae. Methods To isolate the compounds by silica gel and Sephadex LH-20 column chromatography and to elucidate their structures by means of spectral analyses. Results Eight triterpenoids were obtained from EtOAc fraction of EtOH extract and identified as ganoderiol A (Ⅰ), ganodermanontriol (Ⅱ), ganodermatriol (Ⅲ), ganoderic acid C (Ⅳ), ganoderic acid A (Ⅴ), lucidone A (Ⅵ), lucidenic acid C (Ⅶ), and lucidenic acid LM1 (Ⅷ). Conclusion Compounds Ⅰ-Ⅷ are all isolated from G. tsugae for the first time.