Pharmacodynamics of frigid zone plant Taxus cuspidata S.et Z.against skin melanin deposition,oxidation,inflammation and allergy

Background:Taxus cuspidata S.et Z.is a precious species of frigid zone plant belonging to the Taxaceae family,which possesses anticancer,anti-inflammatory,hypoglycemic,and antibacterial pharmacological properties.While taxane extracted from Taxus chinensis has been reported to elicit antioxidant activities,whether Taxus cuspidata S.et Z.has skin-protective actions against injuries remained unknown.This study aims to explore the pharmacological effects of three Taxus extracts on skin melanin deposition,oxidation,inflammation,and allergy so as to provide new ideas for the prevention and treatment of various diseases related to skin damage.Methods:Skin melanin deposition was evaluated by measuring melanin content in the skin of guinea pigs by alkali lysis method.Antioxidant capacity was evaluated by measuring superoxide dismutase(SOD)concentration and glutathione(GSH)content in skin tissue homogenates of Kunming mice by SOD assay kit and micro reduced GSH assay kit.The quantitative real-time polymerase chain reaction(qRT-PCR)and western blotting were used to examine the levels of both SOD and recombinant glutathione peroxidase 4(GPX4).Skin inflammation was evaluated by xylene-induced ear swelling test and egg-white-induced paw swelling test in mice.In a mouse model of skin allergy induced by 4-aminopyridine(4-AP),allergy was determined by licking body counts and histamine concentrations in tissue homogenates using enzyme-linked immunosorbent assay(ELISA)kits.Two proinflammatory factors tumor necrosis factor(TNF)-αand interleukin(IL)-1βwere measured by qRT-PCR.Hematoxylin and eosin(HE)staining was conducted to assess the degree of skin lesion.Results:All three Taxus extracts including Taxus chinensis essential oil,Taxus chinensis extract and Taxus chinensis extract compound reduced the melanin deposits in the back skin relative to the non-treated control animals,of which Taxus chinensis essential oil produced the greatest effect.In contrast,the three Taxus extracts elevated SOD and GSH levels in the skin tissues,and the highest increase was seen with Taxus chinensis essential oil.Three Taxus extracts,especially Taxus chinensis essential oil,effectively reduce the rate of ear and paw swelling.All three Taxus extracts reduced the number of body licks,the levels of TNF-αand IL-1β,and the histamine content in tissue homogenates of mice and alleviated skin damage.Consistently,Taxus chinensis essential oil yielded the greatest magnitude of decreases.Conclusion:While all three Taxus extracts possessed the anti-skin melanin deposition,oxidation,and allergy properties,Taxus chinensis essential oil produced the superior effects.

Population structure and distribution pattern of Taxus cuspidata in Muling region of Heilongjiang Province, China

An investigation was conducted on distribution pattern, site condition and population structure of yew Taxus cuspidata Sieb. et Zucc. in Muling Forest Bureau of Heilongjiang Province, China in April, 2005. Results showed that yew is mainly distributed under the main storey of natural mixed forest of conifer and broadleaf, the soil moisture content of the yew site is high （40%-60%）, the pH value of soil is relatively lower （4.7-5.5）, and that the population structure of wild yew is not rational, belonging to the degeneration population, which is one of the reasons leading to the population decline. Although the site conditions of Muling area are suitable for the growth of wild yew, the population of wild yew shows a decline tendency, due to the fact that the middle-sized adult yew trees have been cut, young yews are often grazed by wildlife, and that the trunks of adult yew tend to be hollow.

Effect of elicitors, precursors and metabolic inhibitors on paclitaxel production by Taxus cuspidata cell culture

In this study, using Taxus cuspidata as a raw material, we obtained stable high-yielding cell lines by subculturing and quantified paclitaxel content using ultra- sonic extraction combined with TLC-UV spectrophotom- etry. In single factor and multiple factors tests to optimize design and study the effects of elicitors, precursors, and metabolic inhibitors on paclitaxel production by Taxus cuspidata cells, paclitaxel production reached 4.32 mg/L when 100 μmol/L methyl jasmonate, 20 mg/L salicylic acid, 400 mg/L phenylalanine and 2 mg/L gibberellin （GA3） were added to the culture medium of suspension cells. When adding metabolic adjustment factors on the 7th day of culture, extra- and intracellular paclitaxel production was the highest at 4.855 mg/L, paclitaxel release rate was 10.48 %, fresh mass and paclitaxel production of cell increased, respectively, by 6.08 and 11.57 %. By controlling the anabolism of paclitaxel, paclitaxel yield was significantly improved.

Ce^(4+)-Induced Apoptosis of Taxus cuspidata Cells in Suspension Culture

The standard detection hallmarks of apoptosis of Taxus cuspidata cells in suspension culture with Ce 4+ were studied. The condensation and margination of chromatin were observed under the electron microscopy. DNA fragmentation ranged 'DNA ladder' on agarose gel electrophoresis. TdT mediated dUTP nick end labeling (TUNEL) analysis of the cells reveals that the nuclear DNA strand breaks can be identified by labeling free 3′ OH termini. These results suggest that Ce 4+ can induce apoptosis of Taxus cuspidata cells and also indicate that there is a certain relationship between apoptosis and secondary metabolite product Taxol.

The selection and stability analysis of stable and high Taxol-producing cell lines from Taxus cuspidata

In this study to screen for stable, high Taxolproducing cell lines（CL5, CL12, and CL21） of Taxus cuspidata, stem tissues were used to induce calli, which were then subcultured nine times to establish suspension cell cultures. From 97 cell lines obtained from conditioned cultures, 10 cell lines with high Taxol content were selected. Stability analyses on solid and liquid B5 media were then used to obtain lines that stably produced high levels of Taxol. Fresh biomass and Taxol production of the ninth generation became stable. Taxol content of selected CL5, CL12, and CL21 samples was 0.0448, 0.0477, and0.0428% of dry mass（DW）, respectively. Proliferation of CL5, CL12 and CL21 was 346.3, 382.5, and 409.2%,respectively. From work over about 2 years, the three cell lines appear suitable for mass production of Taxol,promoting the industrialisation and commercial-scale production of Taxol using cell culture.

A PRELIMINARY STUDY ON THE PHENOLOGY OF TAXUS CUSPIDATA SIEB.ET ZUCC.

In this paper, the phenomena of the life cycle of Taxus cuspidata Sieb. et Zucc., such as germination, leafing, flowing seed mature,new shoot growth etc. were observed and measured. As a result, the phenological regularity of Taxus cuspidata was found initially. At the same time,according to its five-day mean increment and height growth curve, the growing process of new shoot of Taxus cupidata was divided into three periods:slow period peak period and decline period. The paper supplied scientific data for the introduction of the rare and imininent variety, domestication,cultivating, expoiting, utilizing and protecting of Taxus cuspdata.

Studies on Factors Affecting the in vitro Germination and Growth of Taxus cuspidata Embryos

Factors affecting the in vitro germination and growth of Taxus cuspidata embryos were examined. DCR medium was the best among 6 basal media tested; embryos from stage II seeds were the optimal developmental stage for in vitro germination; as the seeds approached maturity, germinability of embryos decrease. Embryos cultured in darkness tended to form callus and decrease the frequency of germination; inoculation method obviously affected frequency of embryo germination. Embryos cultured in optimal conditions developed into seedlings in 2 months.

Metabolites isolated from Penicillium HDS-Z-1E, an endophytic fungal strain isolated from Taxus cuspidata and their activation effect of catalase

Objective:To study the compounds isolated from Penicillium HDS-Z-1E,an endophytic fungal strain isolated from Taxus cuspidata and their activation effect of catalase(CAT).Methods:The chemical constituents were isolated from Penicillium HDS-Z-1E,by using silica gel,Sephadex LH-20 and HPLC.The structural elucidations of five metabolites were elucidated on the basis of spectroscopic including 1 H-NMR,13C-NMR,HMBC and HSQC.Their activation sites of catalase have been investigated by molecular docking.Results:Five metabolites,compounds(1–5)were isolated from Penicillium HDS-Z-1E and identified as 4-hydroxy-4-methyltetrahydro-2H-pyran-2-one(1),4-hydroxymethyl-5,6-dihydro-pyran-2-one(2),5,6-dihydro-2-oxo-2H-pyran-4-carboxylic(3),N-acetyl-hydrazinobenzoic acid(4),and methyl 2-(2,5-dihydroxyphenyl)acetate(5).Conclusion:Compound 3 is a new compound.Compounds 3 and 4 may have potential activators of catalase,providing a theoretical basis for the development of CAT activators.

Effect of La,Ce on Taxus Cuspidata Cell Growth,Biosynthesis and Release of Taxol

The effect of La, Ce was firstly tested on growth of the Taxus(T.) cuspidata cell suspensions, biosynthesis and release of taxol. The results indicate that the growth pattern of T.cuspidata cells is altered significantly by adding high concentration of rare earth in the medium. The lag and exponential phases of cell growth are shortened, the stationary phase disappears and the biomass fluctuates periodically during the decline phase. The rare earth compounds added in the exponential phase obviously increase the taxol biosyntheis and release yields of T.cuspidata cells, and the supplement of carbon source in the medium containing rare earth is also favorable to taxol biosynthesis.