Cephalotaxus Sinensis Research Articles

Effects of Different Treatments on Physiological Characteristics of <i>Cephalotaxus sinensis</i> Seeds

Cephalotaxus sinensis seeds can’t germinate even in the appropriate environment. However, numerous studies have showed that cold stratification and gibberellin acid (GA) can break the seed dormancy and promote seed germination effectively. To investigate the effect of cold stratification and different concentrations of exogenous GA3 on dormancy breaking in seeds of Cephalotaxus sinensis, we monitored germination rates and changes in soluble sugar, starch, amylase, soluble protein, free amino acid during cold stratification. The results showed that seeds stratified for 5 months germinated to 12.7%, while those disposed with 200, 400, 600 mg/L of GA3 germinated to 29.2%, 21.7%, and 18.4%, respectively. Free amino acid content was enhanced significantly, whereas soluble sugar content decreased during 45 days and then increased constantly. Additionally, the main reserves such as starch, protein decreased significantly during cold stratification, and cold stratification induced increases in the activities of α-amylase, (α + β)-amylase. The preliminary results show that the combination of GA3 and cold stratification has better effect to break seed dormancy.

The Causes of Dormancy and the Changes of Endogenous Hormone Content in <i> Cephalotaxus sinensis </i>Seeds

In many wild species, seeds are dormancy at maturity and will not germinate even under favorable environment conditions. Dormancy is a complex trail that is determined by many factors. Some studies have shown that cold stratification and the application of gibberellic acid (GA) can break seed dormancy and promote seed germination. The present study investigated the causes of plant dormancy and the effect of cold stratification and different concentrations of exogenous GA3 in regulating Cephalotaxus sinensis seed germination. Results showed that C. Sinensis seeds have good water permeability, which suggested that seed coats were not the main cause that inhibited the seed germination. There were germination inhibitions in all parts of seeds, and the order of inhibitory effect was: testa < endosperm (embryo), which indicated that existence of germination inhibitions was the main reason causing seed dormancy. Endogenous GAs and IAA (indole-3-acetic acid) content increased, while ABA (abscisic acid) content decreased over the experiments. ZR (zeatin riboside) content decreased in the early phase of cold stratification, but rebounded by the end of the experimental period. The changes of endogenous hormone indicated that GA, IAA and ZR played a positive role in seed germination, whereas ABA was associated with seed dormancy. Besides, the relative ratio of GA/ABA, IAA/ABA and ZR/ABA may play a more important role than their absolute level during the seed development.

Assessing the Capacity of Plant Species to Accumulate Particulate Matter in Beijing, China.

Air pollution causes serious problems in spring in northern China; therefore, studying the ability of different plants to accumulate particulate matter (PM) at the beginning of the growing season may benefit urban planners in their attempts to control air pollution. This study evaluated deposits of PM on the leaves and in the wax layer of 35 species (11 shrubs, 24 trees) in Beijing, China. Differences in the accumulation of PM were observed between species. Cephalotaxus sinensis, Euonymus japonicus, Broussonetia papyriferar, Koelreuteria paniculata and Quercus variabilis were all efficient in capturing small particles. The plants exhibiting high amounts of total PM accumulation (on leaf surfaces and/or in the wax layer), also showed comparatively high levels of PM accumulation across all particle sizes. A comparison of shrubs and trees did not reveal obvious differences in their ability to accumulate particles based on growth form; a combination of plantings with different growth forms can efficiently reduce airborne PM concentrations near the ground. To test the relationships between leaf traits and PM accumulation, leaf samples of selected species were observed using a scanning electron microscope. Growth forms with greater amounts of pubescence and increased roughness supported PM accumulation; the adaxial leaf surfaces collected more particles than the abaxial surfaces. The results of this study may inform the selection of species for urban green areas where the goal is to capture air pollutants and mitigate the adverse effects of air pollution on human health.

Hypoglycemic and hypolipidemic effects of Cephalotaxus sinensis in STZ-induced diabetic rats

Hypoglycemic and hypolipidemic activities of Cephalotaxus sinensis extracts were studiedin streptozotocin-induced diabetic rats. The animals were divided into normal control, diabetic control, diabetic treated and control treated group (n = 6). Effect of oral administration of 80% aqueous ethanolic extract (aq.EE), water extract (WtE), ethyl acetate fraction (EaF), and butanol fraction (BtF) of C. sinensis (200 mg/kg) based on the body weight for 28 days, level of blood glucose, total cholesterol (TC), triglycerides (TG), low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C) were determined. It was found that the ethyl acetate fraction of C. sinensis significantly lowered the elevated blood glucose level, cholesterol, triglycerides, LDL-C, and it showed a significant increase in body weight and HDL-C level (p<0.01). These results obviously indicated that C. sinensis possesses promising hypoglycemic and hypolipidemic effects, which could exert a beneficial action against high glucose level as well as reduced effect on HDL. Key words: Cephalotaxus sinensis, hypoglycemic, hypolipidemic.

Characterization of flavone glycosides and aglycones in Cephalotaxus sinensis by HPLC-DAD-MS

High performance liquid chromatography diode array detector electrospray ionization mass spectrometry (HPLC-DAD-ESI-MS) was utilized to separate and identify flavone glycosides and aglycones in Cephalotaxus sinensis, a natural plant with an anti-hyperglycemic effect. The HPLC-DAD-ESI-MS system could provide a great amount of information on the structure of flavone aglycones and glycosides in C. sinensis without time-consuming pre-separation or purification procedures. A total of seven flavone glycosides and two flavone aglycones were unambiguously identified based on their retention times, ultraviolet spectra, molecular weights and MS/MS spectra. It's interesting that five 5-O-flavone glycosides were found in C. sinensis since 5-O-flavone glycosides are rare and difficult to form in the natural plant. Among them, quercitrin and apigenin-5-O-β-D-glucopyranoside were isolated from this genus for the first time. A novel flavone glycoside, chryoseriol-5-O-[α-L-rhamnopyranosyl-(1→4)-β-D-6-O-acetylglucopyranoside], was isolated for the first time and identified by the ultraviolet spectrum, infrared spectrum, mass spectrum and nuclear magnetic resonance spectrum. In addition, the proposed fragment pathway of the novel flavone glycoside was concluded in the negative ionization mode by HPLC-ESI-MS/MS.

Abstract 1534: Bioassay-guided identification of novel P-glycoprotein inhibitors from traditional Chinese medicines

Abstract Multi-drug resistance (MDR) is an obstacle for therapeutic success of anticancer agents. Overexpression of P-glycoprotein (P-gp), encoded by ABCB1 gene, plays a key role in conferring MDR via active efflux of anticancer drugs from cells. Previous efforts with several generations of P-gp inhibitors had not been successful in modulating clinical MDR. Recently, an increasing number of cancer patients are using complementary and alternative medicines, in particular, traditional Chinese medicines (TCMs), in combination with conventional anticancer agents. The purpose of this study is to examine the effects of TCM extracts on modulating P-gp function. We initiated our study from four single herbal products (Glycyrrhiza uralensis, Hibiscus syriacus, Tripterygium wilfordii, and Cephalotaxus sinensis) and one formula of herbal mixture (Si-Wu-Tang), using a bio-assay guided fractionation method. Crude extracts were firstly prepared with ethanol. The most active extracts identified were further extracted with hexane, chloroform, acetone and ethanol sequentially to obtain fractions of increasing polarity. The MDR modulating effects of the crude extracts and individual fractions were evaluated based on their capability to enhance daunorubicin (DNR) accumulation in P-gp over-expressing human leukemia cell line K562/DOX. The intracellular fluorescence intensity in the presence or absence of test drugs or known P-gp inhibitor cyclosporine A (10 µM) was measured by flow cytometer. The crude extracts were also tested for cytotoxicity against K562/DOX cells by the MTS cell proliferation assay. This study revealed that among the five TCMs, the crude extracts of Tripterygium wilfordii demonstrated a dose-dependent effect on increasing the DNR accumulation, indicating the presence of novel P-gp inhibitor(s). Upon further separation, the chloroform fractions of this herb showed the highest P-gp inhibitory activity. However, known compounds present in this herb, celastrol, triptolide and myristic acid did not show effects on DNR accumulation. Most of the plant extracts tested were able to inhibit the growth of K562/DOX cells with Cephalotaxus sinensis being the most effective, consistent with literature reports that this herb contains highly cytotoxic compound homoharringtonine and congeners. The potency of P-gp inhibition for the five TCMs was not correlated with their cytotoxicity. In conclusion, our results demonstrated that the chloroform fractions of Tripterygium wilfordii contain unknown components for P-gp inhibition. Our current efforts are focused on isolation and identification of active compounds from it. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 1534.

Attenuation of Biochemical Parameters in Streptozotocin-induced Diabetic Rats by Oral Administration of Extracts and Fractions of &lt;i&gt;Cephalotaxus sinensis&lt;/i&gt;

Cephalotaxus sinensis (C. sinensis) large size, evergreen tree common in China and utilized for numerous effective pharmacological applications in Chinese traditional medicine. The hepato-renal effects of C. sinensis were evaluated in vivo using Streptozotocin (STZ)-induced diabetic rats as an tentative model. Animals were orally treated with 80% EtOH extract (aq.EE), H2O extract (WtE) and ethylacetate (EaF)/butanol fractions (BtF) of C. sinensis (200 mg/kg, b.w.) for 28 days whereas control received vehicle merely. The degree of fortification was measured by using biochemical parameters like serum transaminases (ALT and AST), alkaline phosphatase (ALP), creatinine, urea and urine sugar. Meanwhile, the histopathological studies were conducted out to support the above parameters. Administration of C. sinensis aq.EE/BtF (p<0.05) and EaF (p<0.01) patently prevented STZ-induced elevation levels of serum ALT, AST, ALP, creatinine, urea, urine sugar and increase body weight respectively, which were comparable with the standard drug tolbutamide, while WtE did not show any significant effect (p>0.05). Phytochemical studies revealed the presence of saponins, terpenes, sterols and flavonoids in C. sinensis which could be responsible for the possible hepato-renal protective action. The results sustain the fact that the extract/fractions of C. sinensis have an immense potential to be developed further into a phytomedicine.

Attenuation of Biochemical Parameters in Streptozotocin-induced Diabetic Rats by Oral Administration of Extracts and Fractions of Cephalotaxus sinensis.

Cephalotaxus sinensis (C. sinensis) large size, evergreen tree common in China and utilized for numerous effective pharmacological applications in Chinese traditional medicine. The hepato-renal effects of C. sinensis were evaluated in vivo using Streptozotocin (STZ)-induced diabetic rats as an tentative model. Animals were orally treated with 80% EtOH extract (aq.EE), H2O extract (WtE) and ethylacetate (EaF)/butanol fractions (BtF) of C. sinensis (200 mg/kg, b.w.) for 28 days whereas control received vehicle merely. The degree of fortification was measured by using biochemical parameters like serum transaminases (ALT and AST), alkaline phosphatase (ALP), creatinine, urea and urine sugar. Meanwhile, the histopathological studies were conducted out to support the above parameters. Administration of C. sinensis aq.EE/BtF (p<0.05) and EaF (p<0.01) patently prevented STZ-induced elevation levels of serum ALT, AST, ALP, creatinine, urea, urine sugar and increase body weight respectively, which were comparable with the standard drug tolbutamide, while WtE did not show any significant effect (p>0.05). Phytochemical studies revealed the presence of saponins, terpenes, sterols and flavonoids in C. sinensis which could be responsible for the possible hepato-renal protective action. The results sustain the fact that the extract/fractions of C. sinensis have an immense potential to be developed further into a phytomedicine.

Two new flavonoid diglycosides from Cephalotaxus sinensis

Two new acetylated flavonoid diglycosides, scutellarein-5- O-[α- l-rhamnopyranosyl-(1 → 4)-β- d-6- O-acetylglucopyranoside] ( 1), apigenin-5- O-[α- l-rhamnopyranosyl-(1 → 2)-β- d-3,6-di- O-acetylglucopyranoside] ( 2), were isolated from the leaves of Cephalotaxus sinensis. Their structures were determined by chemical and spectroscopic methods including HR-FAB-MS, 1H– 1H COSY, HSQC and HMBC experiments.

Chromatographic fingerprint analysis of Cephalotaxus sinensis from various sources by high-performance liquid chromatography–diodearray detection–electrospray ionization-tandem mass spectrometry

Selective and efficient analytical methods are required not only for quality assurance but also for authentication of Chinese herbal medicine. A simple, rapid and valid fingerprint method has been first carried out for the quality control of Cephalotaxus sinensis by using high-performance liquid chromatography (HPLC) coupled with photodiode array detection (DAD) and electrospray ionisation-tandem mass spectrometry (ESI-MS). The characteristic analytical fingerprints of this plant extract showed 18 common peaks, and out of these, 10 compounds involving 2 new compounds were identified by comparing the retention time, UV and ESI-MS/MS spectrum of each standard with those of each peak separated by on-line HPLC–DAD–MS/MS. Moreover, the effects of collecting locations, harvesting time, storage time, drying methods, and medicinal portions on herbal chromatographic fingerprints were examined by similarity analysis and principal component analysis (PCA) along with markers. Using the reference fingerprint along with markers, the best harvesting time, cultivation location and medicinal part were determined. The results obtained suggest that the chromatographic fingerprint combining similarity evaluation and PCA along with markers or pharmacologically active constituents can efficiently identify raw herb of Cephalotaxus sinensis from different sources, which provide helpful clues to the study of plant's secondary metabolites and benefit quality control.

Antihyperglycemic Effect of Cephalotaxus sinensis Leaves and GLUT-4 Translocation Facilitating Activity of Its Flavonoid Constituents

The objectives of this study were to investigate the antihyperglycemic effect of Cephalotaxus sinensis leaves and to identify the active components. The antihyperglycemic effect of various fractions (FA, FB, FC, FD) of the 80% ethanol extract of the leaves was evaluated in streptozotocin (STZ)-induced diabetic rats. Among the tested fractions, FC was the most active. FC (0.48 g/kg) given orally for 10 d reduced significantly (p<0.001) the blood glucose of STZ-induced diabetic rats. The food and water intakes of FC (0.48 g/kg)-treated diabetic rats were reduced significantly (p<0.001) when compared to the 0.5% carboxymethyl cellulose (CMC)-treated diabetic rats. The activity-guided fractionation of the ethanol extract of C. sinensis leaves furnished three flavonoid compounds, apigenin-5-O-[alpha-L-rhamnopyranosyl-(1-->4)-6-O-beta-D-acetylglucopyranoside] (1), apigenin (2), and apigenin-5-O-[alpha-L-rhamnopyranosyl-(1-->4)-6-O-beta-D-glucopyranoside] (3). The elevation of GLUT-4 protein level in membrane preparations from mice adipocytes was detected by Western blot analysis after adipocytes were pre-incubated with FC (0.1, 1, 10 mg/ml), apigenin (0.1, 2 mg/ml) and apigenin-5-O-[alpha-L-rhamnopyranosyl-(1-->4)-6-O-beta-D-acetylglucopyranoside] (0.1, 2 mg/ml), respectively. Phytochemical investigation and HPLC-DAD analysis of FC indicated that the flavonoids were the major constituents in this fraction. These results suggest that the fraction from C. sinensis leaves is a promising drug for the treatment of diabetes, and that the flavonoids from this plant are the active constituents.