Taxus cuspidata,a rare species of the Taxus genus,and its wild resources are under severe threat.The development of cultivated species has become an important strategy to replace wild species.The objective of this wor...Taxus cuspidata,a rare species of the Taxus genus,and its wild resources are under severe threat.The development of cultivated species has become an important strategy to replace wild species.The objective of this work was to elucidate the differences in secondary metabolite accumulation,particularly in the paclitaxel biosynthesis pathway,between wild and cultivated species.This study employed liquid chromatography-tandem mass spectrometry(LC-MS/MS)and RNA sequencing(RNA-Seq)technologies to conduct integrated metabolomic and transcriptomic analyses of wild and cultivated species of T.cuspidata.The results showed that the content of paclitaxel in cultivated species was significantly higher than in wild species,reaching 1.67 times that of the latter(p 0.01).Additionally,the<content of key paclitaxel precursors,GGPP and 10-deacetylbaccatin III,in cultivated species was 1.94 times(p 0.05)<and 1.71 times(p 0.01)higher than in wild species,respectively.Transcriptomic analysis identified 2606 differentially<expressed genes(DEGs),among which key enzyme genes related to paclitaxel biosynthesis(such as DXS,DXR,GGPS,etc.)were generally upregulated in cultivated species.Multiple key enzyme genes in both the 2-C-methyl-D-erythritol 4-phosphate pathway(MEP)and paclitaxel biosynthesis pathways were significantly upregulated in cultivated species.Conversely,genes and metabolites related to sugar metabolism were found to be higher in content in wild species.These findings reveal the significant advantage of cultivated species in paclitaxel production capacity,providing new insights into the metabolic regulation mechanisms during yew domestication.This has important implications for optimizing paclitaxel biosynthesis and guiding future improvements in T.cuspidata cultivars.展开更多
In order to understand the role of active oxygen species in mediating plant injuries induced by far-UV radiation, seedlings of Taxus cuspidata Sieb. et Zucc. were irradiated by far-UV rays in laboratory for 4 weeks. T...In order to understand the role of active oxygen species in mediating plant injuries induced by far-UV radiation, seedlings of Taxus cuspidata Sieb. et Zucc. were irradiated by far-UV rays in laboratory for 4 weeks. The production of organic free-radicals in detached needles, and the production of O-2(radical anion) and O-1(2) in isolated chloroplasts were detected weekly by electron spin resonance (ESR) to evaluate their relative importance. The results show that the cumulative effect of far-UV irradiation, is best indicated by the production of organic free radicals in the needles, O-2(radical anion) production in chloroplasts is the next. The enhancement of O-1(2) production in chloroplasts by the cumulative far-UV irradiation seems to be not so important as O-2(radical anion) in mediating injuries induced by, far-UV radiation because of its high background value.展开更多
[Objective] This study aimed to investigate the browning of T. cuspidata cells in suspension culture and provide the guidance for the cell suspension culture of T. cuspidata. [Method] T. cuspidata callus was used as e...[Objective] This study aimed to investigate the browning of T. cuspidata cells in suspension culture and provide the guidance for the cell suspension culture of T. cuspidata. [Method] T. cuspidata callus was used as experimental materials, to explore the effect of different medium, N/P ratio, pH, shaking speed, illumination time and light intensity and other factors on browning of T. cuspidata cells in suspension culture. [Result] Non-browning callus was transferred to 2MB5 medium （pH 7.0） for illumination culture at 22℃ under light intensity of 1 500 lx with shaking speed of 90 r/min for 24 h. Results showed that the cell browning was significantly inhibited. [Conclusion] This study laid the foundation for cell suspension culture of T. cuspidata and had important significance to the large-scale industrial production of paclitaxel.展开更多
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 sho...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.展开更多
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 si...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.展开更多
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- e...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.展开更多
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 regul...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.展开更多
Taxus cuspidata is a rare plant with important medicinal and ornamental value.Aiming at the obvious differences between wild and cultivated populations of T.cuspidata from Northeast China,a total of 61 samples,that is...Taxus cuspidata is a rare plant with important medicinal and ornamental value.Aiming at the obvious differences between wild and cultivated populations of T.cuspidata from Northeast China,a total of 61 samples,that is,33 wild yews and 28 cultivated yews were used to analyze the differences and correlations of the kinship,genetic diversity,and genetic structure between them by specific length amplified fragment sequencing(SLAF-seq).Finally,470725 polymorphic SLAF tags and 58622 valid SNP markers were obtained.Phylogenetic analysis showed that 61 samples were classified into 2 clusters:wild populations and cultivated populations,and some wild yews were categorized into the cultivated populations;the genetic diversity analysis showed that the Nei diversity index of wild populations(0.4068)was smaller than that of cultivated populations(0.4414),and the polymorphic information content(PIC)of wild populations(0.2861)was smaller than that of cultivated populations(0.3309).The genetic differentiation analysis showed that the total populations of gene diversity(H_(t))of cultivated and wild populations were respectively 0.8159 and 0.5685,the coefficient of gene differentiation(G_(st))of cultivated and wild populations was respectively 0.3021 and 0.1068,and the gene flow(N_(m))(2.4967)of wild populations was larger than cultivated populations(0.8199).The molecular variance(AMOVA)revealed that inter-population variation accounted for 29.57%of the total genetic variation,while intra-population variation accounted for 70.42% of the total genetic variation(p<0.001),this suggested that the genetic variation in the T.cuspidata is mainly attributed to within-population factors.In conclusion,the genetic distance between geographical ecological groups of wild populations was generally smaller than that of cultivated populations,and the degree of genetic diversity and genetic differentiation was smaller than that of cultivated populations.As evident,the utilization of SLAF-seq technology enables efficient and accurate development of SNP markers suitable for genetic analysis of T.cuspidata species.These developed SNP markers can provide a molecular foundation for T.cuspidata breeding,construction of genetic maps,variety identification,and association analysis of agronomic traits.展开更多
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....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.展开更多
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...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.展开更多
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.Whil...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.展开更多
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 ...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.展开更多
基金supported by grants from the National Science Foundation of China to Yanwen Zhang(32272757,31972363)grants from Liaoning Provincial Department of Education Project to Dandan Wang(JYTMS20230698)grants from the Liaoning Provincial Science and Technology.Fund Project to Dandan Wang(2023JH2/101700200).
文摘Taxus cuspidata,a rare species of the Taxus genus,and its wild resources are under severe threat.The development of cultivated species has become an important strategy to replace wild species.The objective of this work was to elucidate the differences in secondary metabolite accumulation,particularly in the paclitaxel biosynthesis pathway,between wild and cultivated species.This study employed liquid chromatography-tandem mass spectrometry(LC-MS/MS)and RNA sequencing(RNA-Seq)technologies to conduct integrated metabolomic and transcriptomic analyses of wild and cultivated species of T.cuspidata.The results showed that the content of paclitaxel in cultivated species was significantly higher than in wild species,reaching 1.67 times that of the latter(p 0.01).Additionally,the<content of key paclitaxel precursors,GGPP and 10-deacetylbaccatin III,in cultivated species was 1.94 times(p 0.05)<and 1.71 times(p 0.01)higher than in wild species,respectively.Transcriptomic analysis identified 2606 differentially<expressed genes(DEGs),among which key enzyme genes related to paclitaxel biosynthesis(such as DXS,DXR,GGPS,etc.)were generally upregulated in cultivated species.Multiple key enzyme genes in both the 2-C-methyl-D-erythritol 4-phosphate pathway(MEP)and paclitaxel biosynthesis pathways were significantly upregulated in cultivated species.Conversely,genes and metabolites related to sugar metabolism were found to be higher in content in wild species.These findings reveal the significant advantage of cultivated species in paclitaxel production capacity,providing new insights into the metabolic regulation mechanisms during yew domestication.This has important implications for optimizing paclitaxel biosynthesis and guiding future improvements in T.cuspidata cultivars.
文摘In order to understand the role of active oxygen species in mediating plant injuries induced by far-UV radiation, seedlings of Taxus cuspidata Sieb. et Zucc. were irradiated by far-UV rays in laboratory for 4 weeks. The production of organic free-radicals in detached needles, and the production of O-2(radical anion) and O-1(2) in isolated chloroplasts were detected weekly by electron spin resonance (ESR) to evaluate their relative importance. The results show that the cumulative effect of far-UV irradiation, is best indicated by the production of organic free radicals in the needles, O-2(radical anion) production in chloroplasts is the next. The enhancement of O-1(2) production in chloroplasts by the cumulative far-UV irradiation seems to be not so important as O-2(radical anion) in mediating injuries induced by, far-UV radiation because of its high background value.
基金Supported by National Natural Science Foundation of China (31070164)Young Scientists Fund of Dalian (2006J23JH031)~~
文摘[Objective] This study aimed to investigate the browning of T. cuspidata cells in suspension culture and provide the guidance for the cell suspension culture of T. cuspidata. [Method] T. cuspidata callus was used as experimental materials, to explore the effect of different medium, N/P ratio, pH, shaking speed, illumination time and light intensity and other factors on browning of T. cuspidata cells in suspension culture. [Result] Non-browning callus was transferred to 2MB5 medium （pH 7.0） for illumination culture at 22℃ under light intensity of 1 500 lx with shaking speed of 90 r/min for 24 h. Results showed that the cell browning was significantly inhibited. [Conclusion] This study laid the foundation for cell suspension culture of T. cuspidata and had important significance to the large-scale industrial production of paclitaxel.
文摘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.
文摘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.
基金supported by development plan project during ‘‘the 12th Five Year Plan’’ Nation Science and Technology in rural area(No.2012AA10A506-04 and No.2013AA103005-04)Changchun City science and technology development program(No.2014174)Changchun City science and technology support program(No.2014NK002)
文摘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.
文摘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.
基金This work was supported by Grants from the National Science Foundation of China to Yanwen Zhang(32272757,31972363)Grants from the Liaoning Provincial Department of Education Project to Dandan Wang(JYTMS20230698)Grants from the Liaoning Provincial Science and Technology Fund Project:Comparative Multi-Omics Study of Wild and Cultivated Species of Taxus chinensis.
文摘Taxus cuspidata is a rare plant with important medicinal and ornamental value.Aiming at the obvious differences between wild and cultivated populations of T.cuspidata from Northeast China,a total of 61 samples,that is,33 wild yews and 28 cultivated yews were used to analyze the differences and correlations of the kinship,genetic diversity,and genetic structure between them by specific length amplified fragment sequencing(SLAF-seq).Finally,470725 polymorphic SLAF tags and 58622 valid SNP markers were obtained.Phylogenetic analysis showed that 61 samples were classified into 2 clusters:wild populations and cultivated populations,and some wild yews were categorized into the cultivated populations;the genetic diversity analysis showed that the Nei diversity index of wild populations(0.4068)was smaller than that of cultivated populations(0.4414),and the polymorphic information content(PIC)of wild populations(0.2861)was smaller than that of cultivated populations(0.3309).The genetic differentiation analysis showed that the total populations of gene diversity(H_(t))of cultivated and wild populations were respectively 0.8159 and 0.5685,the coefficient of gene differentiation(G_(st))of cultivated and wild populations was respectively 0.3021 and 0.1068,and the gene flow(N_(m))(2.4967)of wild populations was larger than cultivated populations(0.8199).The molecular variance(AMOVA)revealed that inter-population variation accounted for 29.57%of the total genetic variation,while intra-population variation accounted for 70.42% of the total genetic variation(p<0.001),this suggested that the genetic variation in the T.cuspidata is mainly attributed to within-population factors.In conclusion,the genetic distance between geographical ecological groups of wild populations was generally smaller than that of cultivated populations,and the degree of genetic diversity and genetic differentiation was smaller than that of cultivated populations.As evident,the utilization of SLAF-seq technology enables efficient and accurate development of SNP markers suitable for genetic analysis of T.cuspidata species.These developed SNP markers can provide a molecular foundation for T.cuspidata breeding,construction of genetic maps,variety identification,and association analysis of agronomic traits.
文摘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.
基金supported by the ‘‘12th Five Year Plan’’ National Science and Technology in Rural Area(Nos.2013AA103005-04 and 2012AA10A506-04)Changchun City Science and Technology Development Program(No.2014174)+1 种基金Changchun City Science and Technology Support Program(No.2014NK002)Graduate Innovation Fund of Jilin University(No.2016172)
文摘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.
基金This work was supported by the National Natural Science Foundation of Heilongjiang Province-Outstanding Youth Project(No.YQ2020H008).
文摘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.
文摘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.
