RESUMEN

A series of C21 steroidal glycosides were isolated from the root bark of Periploca sepium, including a new compound, perisepiumoside A1 (1), and six known compounds (2-7). Their structures were elucidated by analysis of HR-ESI-MS, and 1D and 2D NMR spectroscopic data. All these compounds were tested for their NO production inhibitory activity in LPS-stimulated RAW 264.7 cells. Results showed that these C21 steroidal glycosides could remarkably inhibit NO production, particularly 1 and 2 with IC50 values of 30.81 ± 0.18 µM and 44.39 ± 0.21 µM, respectively. In addition, the cytotoxicity of these compounds was measured on A549, MCF-7, and HeLa cancer cell lines. Among them, compounds 1 and 7 displayed cytotoxicity against the A549 cell line with IC50 values of 28.41 ± 0.12 µM and 39.06 ± 0.05 µM, respectively.

RESUMEN

Hevea brasiliensis is the main source of natural rubber. Restricted by its tropical climate conditions, the planting area in China is limited, resulted in a low self-sufficiency. Periploca sepium which can produce natural rubber is a potential substitute plant. cis-prenyltransferase (CPT), small rubber particle protein (SRPP) and rubber elongation factor (REF) are key enzymes involved in the biosynthesis of cis-1, 4-polyisoprene, the main component of natural rubber. In this study, we cloned the promoter sequences of CPT, SRPP and REF through chromosome walking strategy. The spatial expression patterns of the three promoters were analyzed using GUS (ß-glucuronidase) as a reporter gene driven by the promoters through Agrobacterium-mediated genetic transformation. The results showed that GUS driven by CPT, SRPP or REF promoter was expressed in leaves and stems, especially in the leaf vein and vascular bundle. The GUS activity in stems was higher than that in leaf. This study provided a basis for analyzing the biosynthesis mechanism of natural rubber and breeding new varieties of high yield natural rubber.

Asunto(s)

Factores de Elongación de Péptidos , Periploca , Factores de Elongación de Péptidos/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Periploca/genética , Periploca/metabolismo , Goma , Fitomejoramiento , Clonación Molecular

RESUMEN

Hevea brasiliensis is the main source of natural rubber. Restricted by its tropical climate conditions, the planting area in China is limited, resulted in a low self-sufficiency. Periploca sepium which can produce natural rubber is a potential substitute plant. cis-prenyltransferase (CPT), small rubber particle protein (SRPP) and rubber elongation factor (REF) are key enzymes involved in the biosynthesis of cis-1, 4-polyisoprene, the main component of natural rubber. In this study, we cloned the promoter sequences of CPT, SRPP and REF through chromosome walking strategy. The spatial expression patterns of the three promoters were analyzed using GUS (β-glucuronidase) as a reporter gene driven by the promoters through Agrobacterium-mediated genetic transformation. The results showed that GUS driven by CPT, SRPP or REF promoter was expressed in leaves and stems, especially in the leaf vein and vascular bundle. The GUS activity in stems was higher than that in leaf. This study provided a basis for analyzing the biosynthesis mechanism of natural rubber and breeding new varieties of high yield natural rubber.

Asunto(s)

Factores de Elongación de Péptidos/genética , Proteínas de Plantas/metabolismo , Periploca/metabolismo , Goma , Fitomejoramiento , Clonación Molecular

RESUMEN

Aberrant microbe-immune cell interaction is a predisposing factor in inflammatory bowel disease (IBD) and colitis-associated cancer (CAC). Cortex Periplocae is a famous traditional Chinese medicine with putative anti-rheumatoid arthritis and anti-dyspepsia effects. Here, we show that the Periploca sepium periplosides (PePs), a cardiac glycosides-free pregnane glycosides extract from root bark of Cortex Periplocae, alleviates colon inflammation, improves intestinal epithelial barrier function, and prevents colitis-associated tumorigenesis in mice with colitis and CAC. Mechanistically, PePs treatment modulates abnormal gut microbiota composition in model mice, especially enriches an anti-inflammatory commensal bacterium A. muciniphila BAA-835. We further demonstrate that the altered gut microbiota following PePs treatment plays an important role in modulation of intestinal Type 17 immunity in both colitis and CAC mouse model. Our results indicate that PePs may be used as a potential gut microbiota modulator to treat IBD and CAC.

Asunto(s)

Neoplasias Asociadas a Colitis , Colitis , Microbioma Gastrointestinal , Enfermedades Inflamatorias del Intestino , Animales , Colitis/complicaciones , Colitis/tratamiento farmacológico , Colitis/microbiología , Sulfato de Dextran , Modelos Animales de Enfermedad , Ratones , Ratones Endogámicos C57BL , Células Th17/patología

RESUMEN

Natural odour compounds could be a potential alternative to synthetic herbicides. The odour compound of Periploca sepium Bunge, named 2-hydroxy-4-methoxy-benzaldehyde (HMB), is a herbicidal compound. However, its herbicidal mechanism is unclear. In this experiment, the physiological and biochemical indices, ultrastructure, and photosynthetic function of the leaves of Humulus scandens (Lour.) Merr. treated by HMB were assessed to elucidate the herbicidal mechanism. The results of physiological and biochemical indices are as follows: First, after 4 h of treatment with 2.5 and 5.0 mg/mL, the damage rates in the membrane permeation assay were 74.7% and 89.1%, respectively. Second, compared to the negative control group, multiple physiological and biochemical indices of the two treated groups were changed, including catalase content (-18.5 and -26.5 ng/mL), superoxide dismutase content (-27.4 and -56.6 ng/mL), peroxidase content (382.0 and 880.0 ng/mL), reactive oxygen species content (16.7 and 27.2 ng/mL), malondialdehyde content (8.9 and 25.2 nmol/g), and water potential values (0.2 and 0.3 MPa), except for the photosynthetic pigment contents (chlorophyll a, b, and carotene). Furthermore, the results of transmission electron microscopy showed that the organelles in the mesophyll tissue cells disappeared and severe plasmolysis led to cell atrophy after 4 h of treatment. There were fewer starch granules after 24 h of treatment, but there was no obvious abnormality in the upper and lower epidermal cells. The results of photosynthetic function showed that in the light response, the net photosynthetic rate (Pn), transpiration rate (Tr), stomatal conductance (Gs), and stomatal limitation value of the tested leaves were lower than those of the negative control group by 26.6 µmol·m-2·s-1, 7.7 mmol·m-2·s-1, 0.9 mol·m-2·s-1, and 0.2, respectively. However, the intercellular CO2 concentration (Ci) increased and was higher than the air CO2 concentration. In the CO2 response, the Pn, Tr and Gs of the tested leaves first increased and then decreased, but the Ci value continuously increased and finally reached 1727.5 µmol·mol-1. It is obvious that HMB may have inhibited the effect on the photosynthetic system of the tested leaves. Overall, HMB killed the weeds by destroying the structure and multiple physiological functions of the tested leaves.

Asunto(s)

Humulus , Periploca , Dióxido de Carbono , Clorofila , Clorofila A , Odorantes , Periploca/fisiología , Fotosíntesis , Hojas de la Planta/fisiología

RESUMEN

The odor compound from Periploca sepium Bunge, 2-hydroxy-4-methoxy-benzaldehyde (HMB), is an allelochemical agent and is one of the least investigated isomers of vanillin. In this study, we used label-free quantitative proteomics analysis technology to investigate the effect of HMB on the protein expression of Humulus scandens (Lour.) Merr. leaves in July 2019 on Guiyang. A total of 269 proteins of 624 identified proteins were differentially expressed, among which 21.18% of the proteins were up-regulated and 32.71% down-regulated. These proteins were classified into 11 cell components and more than 20% of differentially expressed proteins were located in cell membrane and chloroplast. Functional classification analysis showed that 12 molecular functions were altered upon HMB treatment, and the ratio of catalytic activity was the highest (19.53%). At least 12 biological functions were affected, which involved small molecule metabolic processes, organic substance metabolic processes, gene expression, and photosynthesis. Our data provide resources and insights into the biochemical mechanism by which HMB kills weeds.

Asunto(s)

Humulus/fisiología , Odorantes/análisis , Periploca/fisiología , Hojas de la Planta/química , Benzaldehídos , China , Periploca/química , Fotosíntesis , Proteoma/metabolismo , Proteómica

RESUMEN

LC-MS guided chemical investigation of the periploside-rich extract of the root barks of Periploca sepium afforded six new minor pregnane glycosides, named periplosides A1-A6 (1-6). Their structures were characterized on the basis of extensive spectroscopic analysis. Compounds 1-6 were evaluated for their inhibitory activities against the proliferation of T and B lymphocytes in vitro, among them, compound 5 exhibited significant inhibitory activities and the most favorite selective index (SI) values against the proliferation of T lymphocyte (IC50 = 0.30 µM, SI = 176) and B lymphocyte (IC50 = 0.55 µM, SI = 97).

Asunto(s)

Linfocitos B/efectos de los fármacos , Glicósidos/farmacología , Periploca/química , Raíces de Plantas/química , Pregnanos/farmacología , Linfocitos T/efectos de los fármacos , Animales , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Femenino , Glicósidos/química , Glicósidos/aislamiento & purificación , Ratones , Ratones Endogámicos BALB C , Estructura Molecular , Pregnanos/química , Pregnanos/aislamiento & purificación , Relación Estructura-Actividad

RESUMEN

ETHNOPHARMACOLOGICAL RELEVANCE: Periploca sepium Bunge (P. sepium) is used in traditional Chinese medicine (TCM) for the treatment of autoimmune diseases, particularly rheumatoid arthritis. Periploca sepium periplosides (PePs), isolated from the root bark of P. sepium, characterized as the cardiac glycosides-free pregnane glycosides fraction, is expected to possess therapeutic potential on inflammatory arthritis. AIM OF THE STUDY: The current study is designed to evaluate the anti-nociceptive, anti-inflammatory and anti-arthritic activities effects of the PePs. MATERIALS AND METHODS: The anti-nociceptive activity of PePs was examined in the writhing test and hot-plate test in mice. The anti-inflammatory activity of PePs was determined by the 2, 4-dinitro-1-fluorobenzene (DNFB)-induced ear edema model and the carrageenan induced paw edema model in mice. The anti-arthritic activity of PePs was investigated by evaluating the joint inflammation and arthritis pathology in rat adjuvant induced arthritis (AIA) and murine collagen induced arthritis (CIA). Phytohaemagglutinin M (PHA-M) -elicited human peripheral blood mononuclear cells (PBMCs) were further applied to assess the suppressive activity of PePs on IFN-γ and IL-17 production. RESULTS: PePs treatment markedly decreased the acetic acid-induced visceral nociceptive response and increased the hot-plate pain threshold. Further, oral administration of PePs exhibited anti-inflammatory activity by decreasing DNFB-induced ear edema in mice and carrageenan-induced paw edema in rats. Moreover, oral treatment of PePs ameliorated joint swelling and attenuated bone erosion in rodent arthritis, and the therapeutic benefits were partially attributed to the suppression of proinflammatory cytokines such IFN-γ and IL-17. Moreover, PePs suppressed the proliferation as well as IFN-γ and IL-17 secretion in PHA-M-elicited human PBMCs in a concentration dependent manner. CONCLUSIONS: Taken together, our results justified the traditional use of Periploca sepium Bunge for the treatment of diseases associated with inflammation and pain.

Asunto(s)

Analgésicos/farmacología , Antirreumáticos/farmacología , Glicósidos/farmacología , Periploca/química , Pregnanos/farmacología , Analgésicos/aislamiento & purificación , Animales , Antiinflamatorios/aislamiento & purificación , Antiinflamatorios/farmacología , Antirreumáticos/aislamiento & purificación , Artritis Experimental/tratamiento farmacológico , Artritis Experimental/patología , Artritis Reumatoide/tratamiento farmacológico , Artritis Reumatoide/patología , Modelos Animales de Enfermedad , Edema/tratamiento farmacológico , Femenino , Glicósidos/aislamiento & purificación , Inflamación/tratamiento farmacológico , Inflamación/patología , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos DBA , Ratones Endogámicos ICR , Dolor/tratamiento farmacológico , Pregnanos/aislamiento & purificación , Ratas , Ratas Sprague-Dawley

RESUMEN

BACKGROUND: The Periploca sepium bark root (PSBR) has been regarded as a potential botanical insecticide because of its significant insecticidal activity of secondary metabolites. Several periplocosides were isolated from it as promising pesticides to control crop pests in agriculture. RESULTS: In our research, two new periplocosides, along with four known periplocosides were isolated from PSBR. The names of new periplocosides were periplocoside T (PST) and periplocoside U (PSU) while another four periplocosides were known as follows: periplocoside A (PSA), periplocoside F (PSF), periplocoside E (PSE) and periplocoside D (PSD). All periplocosides were evalulated for insecticidal activity against 3rd Mythimna separata (Walker) and Plutella xylostella. The biometric data showed that periplocoside T, PSD and PSF had remarkable insecticidal activity against tested insects. Its values of LD50 were 1.31, 3.94 and 3.42 µg·lavare-1 against 3rd M. separata respectively, while the activity of those compounds against 3rd P. xylostella were 5.45, 12.17 and 13.95 µg·lavare-1 , respectively. It was apparent after further study of the mechanism of action against M. separata was conducted that PST possessed the most significant insecticidal activity. The results of enzymatic activity displayed that powerful activation of tryptase, especially weak alkaline tryptase might be a dominant factor causing death of M. separata in vivo. CONCLUSION: We herein report isolation and the mechanisms of action of insecticidal periplocosides, which established the fundamental development of natural agents to prevent pest damage to crops. © 2020 Society of Chemical Industry.

Asunto(s)

Insecticidas , Mariposas Nocturnas , Periploca , Animales , Insecticidas/farmacología , Dosificación Letal Mediana , Corteza de la Planta

RESUMEN

BACKGROUND: Periploca sepium is traditionally used in Chinese medicine to treat particularly rheumatic disorders and as a tonic. Periplocin was found as the most cytotoxic compound of its root bark and induced death receptor mediated apoptosis in liposarcoma cells. Sarcomas are a rare type of cancer with only a few treatment options. The five-year survival rate of advanced tumors is low. PURPOSE: In this study, we investigated the effects of periplocin in two myxofibrosarcoma (MFS)cell lines, MUG-Myx2a and MUG-Myx2b, which are subclones of the same tumor and reflect the tumor´s heterogeneity, and in T60 primary myxofibrosarcoma cells. METHODS: The xCELLigence system and the CellTiter 96® AQueous assay were used for studying cell viability. FACS and Western blot experiments were used to investigate the effects of periplocin on apoptosis induction, cell cycle distribution, and the expression of cleaved PARP, caspase 3, p53, phospho-histone γH2AX, ERK/phospho ERK, p38/phospho p38, and, finally, JNK/phospho JNK. Additionally, the expression of the apoptotic markers Bim, NOXA, Bak, Bcl-2, Bcl-xl, and the death receptors IGFR, FADD, TRADD, TNFR1A, TRAIL-R1, and TRAIL-R2 were evaluated using reversed real-time PCR. RESULTS: Periplocin decreased dose-dependently the viability of all MFS cell lines and was more effective than the standard chemotherapeutic doxorubicin. It arrested the cells in the G2/M phase and led to caspase activation. Moreover, periplocin increased the mRNA expression of NOXA, Bak, Bcl-2, and death receptors such as TRAIL-R1 and TRAIL-R2 and the protein expression of ERK/phospho ERK, p38/phospho p38, and JNK/phospho JNK. In all cases, differences in the effects in the different subclones were observed. CONCLUSION: Periplocin showed promising effects in MFS cells. The higher effectiveness compared to doxorubicin is an important aspect for further research with regard as a treatment option. The different effects of periplocin in the two subclones showed the great importance of intratumoral heterogeneity in MFS therapy.

RESUMEN

Five new polyphenolic derivatives, sepiumols A-E (1-5), were isolated from the root barks of Periploca sepium. Their structures were elucidated by interpretation of NMR spectroscopic and mass spectrometric data. Compounds 1, 3 and 5 were found to exhibit significant antifungal activity, particularly for 3 with the remarkable activity against Gibberella saubinetii and Alternaria longipes with MIC values of 1.56 and 3.13 µg/mL (ketoconazole: 0.78 µg/mL), respectively. In addition, compounds 1, 3 and 5 also displayed significant antibacterial activity against methicillin-resistant Staphylococcus aureu with MIC values of 12.50-25 µg/mL (ciprofloxacin: 0.78 µg/mL).

Asunto(s)

Antibacterianos/farmacología , Antifúngicos/farmacología , Periploca/química , Polifenoles/farmacología , Alternaria/efectos de los fármacos , Antibacterianos/aislamiento & purificación , Antifúngicos/aislamiento & purificación , Gibberella/efectos de los fármacos , Staphylococcus aureus Resistente a Meticilina/efectos de los fármacos , Estructura Molecular , Fitoquímicos/aislamiento & purificación , Fitoquímicos/farmacología , Corteza de la Planta/química , Raíces de Plantas/química , Polifenoles/aislamiento & purificación

RESUMEN

BACKGROUND: During a screening of Chinese plants traditionally used for the treatment of cancer and related diseases, extracts of the root bark of Periploca sepium Bunge showed strong cytotoxic activity. PURPOSE: Isolate and identify cytotoxic compounds from P. sepium and investigate the effects and mechanism of action on different cancer cell lines. METHODS: Extracts obtained with solvents of different polarities of the root bark of P. sepium were tested for their anti-proliferative effects. The most active extract was subjected to activity-guided fractionation using different chromatographic methods. The most active compound was further investigated on sarcoma cell lines regarding its effects concerning apoptosis, DNA damage and death receptor expression. RESULTS: We isolated the cardiac glycosides periplocin, glucosyl divostroside, periplogenin, periplocymarin and periplocoside M with periplocin exhibiting the lowest IC50 value against leukemia and liposarcoma cells. Liposarcomas are rare tumors within the heterogeneous group of soft tissue sarcomas and respond poorly to conventional treatments. Periplocin led to growth inhibition and apoptosis induction by changing the expression of death receptors and inducing DNA double strand breaks in SW-872 cells. CONCLUSION: Periplocin displays a promising mechanism of action in sarcoma cells because altering the death receptor expression is an interesting target in sarcoma treatment especially to overcome TRAIL resistance.

Asunto(s)

Apoptosis/efectos de los fármacos , Liposarcoma/patología , Periploca/química , Receptores de Muerte Celular/metabolismo , Saponinas/farmacología , Glicósidos Cardíacos , Línea Celular Tumoral , China , Digitoxigenina/análogos & derivados , Humanos , Extractos Vegetales/farmacología , Raíces de Plantas/química , Plantas Medicinales/química

RESUMEN

Periploca sepium Bunge is a traditional medicinal plant, whose root bark is important for Chinese herbal medicine. Its major bioactive compounds are C21 steroids and periplocin, a kind of cardiac glycoside, which are derived from the steroid synthesis pathway. However, research on P. sepium genome or transcriptomes and their related genes has been lacking for a long time. In this study we estimated this species nuclear genome size at 170 Mb (using flow cytometry). Then, RNA sequencing of four different tissue samples of P. sepium (leaves, roots, adventitious roots, and calli) was done using the sequencing platform Illumina/Solexa Hiseq 2,500. After de novo assembly and quantitative assessment, 90,375 all-transcripts and 71,629 all-unigenes were finally generated. Annotation efforts that used a number of public databases resulted in detailed annotation information for the transcripts. In addition, differentially expressed genes (DEGs) were identified by using digital gene profiling based on the reads per kilobase of transcript per million reads mapped (RPKM) values. Compared with the leaf samples (L), up-regulated genes and down-regulated genes were eventually obtained. To deepen our understanding of these DEGs, we performed two enrichment analyses: gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG). Here, the analysis focused upon the expression characteristics of those genes involved in the terpene metabolic pathway and the steroid biosynthesis pathway, to better elucidate the molecular mechanism of bioactive steroid synthesis in P. sepium. The bioinformatics analysis enabled us to find many genes that are involved in bioactive steroid biosynthesis. These genes encoded acetyl-CoA acetyltransferase (ACAT), HMG-CoA synthase (HMGS), HMG-CoA reductase (HMGR), mevalonate kinase (MK), phosphomevalonate kinase (PMK), mevalonate diphosphate decarboxylase (MDD), isopentenylpyrophosphate isomerase (IPPI), farnesyl pyrophosphate synthase (FPS), squalene synthase (SS), squalene epoxidase (SE), cycloartenol synthase (CAS), sterol C-24 methyltransferase (SMT1), sterol-4alpha-methyl oxidase 1 (SMO1), sterol 14alpha-demethylase (CYP51/14-SDM), delta(14)-sterol reductase (FK/14SR), C-8,7 sterol isomerase (HYD1), sterol-4alpha-methyl oxidase 2 (SMO2), delta(7)-sterol-C5(6)-desaturase (STE1/SC5DL), 7-dehydrocholesterol reductase (DWF5/DHCR7), delta (24)-sterol reductase (DWF1/DHCR24), sterol 22-desaturase (CYP710A), progesterone 5beta-reductase (5ß-POR), 3-beta-hydroxysteroid dehydrogenase (3ß-HSD). This research will be helpful to further understand the mechanism of bioactive steroid biosynthesis in P. sepium, namely C21 steroid and periplocin biosynthesis.

RESUMEN

Four new C21 steroidal glycosides (1-4), named perisepiumosides FI (1-4) together with six known steroidal glycosides (5-10) and four oligosaccharides (11-14), were isolated from the root bark of Periploca sepium. Their structures were characterized on the basis of 1D and 2D-NMR spectroscopic data as well as HR-ESI-MS analysis. The evaluation of inhibition activity against human A-549 and HepG2 cell lines indicated that compounds 2, 8, 10 and 13 showed different levels of cytotoxic activities with IC50 values ranging from 0.61 to 7.86µM.

Asunto(s)

Glicósidos/química , Oligosacáridos/química , Periploca/química , Raíces de Plantas/química , Células A549 , Glicósidos/aislamiento & purificación , Células Hep G2 , Humanos , Estructura Molecular , Oligosacáridos/aislamiento & purificación

RESUMEN

Periplocoside NW (PSNW) with pregnane glycoside skeleton is a novel insecticidal compound isolated from the root bark of Periploca sepium Bunge. This compound has a potent stomach poisoning activity against several insect pests. In this study, we observed the intoxication symptoms, investigated the histopathological effects and carried out immuno-electron microscopic localization of PSNW on the midgut epithelium of oriental armyworm Mythimna separata Walker larvae for better understanding its action mechanism against insects. Ultrastructural observations showed that cell damages caused by PSNW in the midgut of M. separata larvae are related to the degeneration of brush border microvilli. The dissolution of cytoskeletal structures in the interior and on the surface of microvilli was responsible for the decrease in size and eventual disappearance of microvilli when bubbles of cytoplasmic substances protrude into the midgut lumen of M. separata, thus resulting in cell death. The immuno-electron microscopic localization research showed that gold particle appeared on the microvilli layer of the midgut of M. separate larvae firstly. The density of gold particle gradually added with the time, and finally microvilli layer was destructed severely. Meantime, the gold particles were also presented to the intracellular organelle membrane and the organelles also were destructed. Therefore, we proposed that this membrane system on insect midgut epithelium cells is the initial acting site of PSNW against insects.

Asunto(s)

Glicósidos/toxicidad , Insecticidas/toxicidad , Mariposas Nocturnas/efectos de los fármacos , Periploca/química , Extractos Vegetales/toxicidad , Pregnanos/toxicidad , Animales , Sistema Digestivo/química , Sistema Digestivo/efectos de los fármacos , Sistema Digestivo/ultraestructura , Epitelio/química , Epitelio/efectos de los fármacos , Epitelio/ultraestructura , Histología , Larva/química , Larva/efectos de los fármacos , Larva/ultraestructura , Microscopía Inmunoelectrónica , Microvellosidades/química , Microvellosidades/efectos de los fármacos , Microvellosidades/ultraestructura , Mariposas Nocturnas/química , Mariposas Nocturnas/ultraestructura

RESUMEN

Objective To investigate the inhibitory effects of periplocin from cortex periplocae (CPP) on human lung cancer cell line QG56 and to discuss its mechanism. Methods QG56 cells were cultured in vitro. The final concentrations of CPP in control group were 1.25, 2.50, 5.00, 10.00 and 20.00μg/L. QG56 cells were treated with ascending concentration of CPP for 24 h, 48 h and 72 h. The cell proliferation was measured using MTT method. The morphological changes of QG56 cells were observed under inverted microscope. Flow cytometry (FCM) was used to detect the effects of CPP on cell cycle and cell apoptosis. The expression of apoptosis associated gene bax mRNA in QG56 cells was detected by RT-PCR. The expres-sion of bax protein before and after treatment of CPP was examined by SP immunocytochemistry. Results The inhibitory ef-fect of CPP on the proliferation of QG56 cells was increased with the increasing concentrations of CPP and the prolonged du-ration of treatment. The morphological changes were displayed in QG56 exposed to CPP. The results of FCM showed that CPP caused cell cycle arrest at G0/G1 phase. The apoptotic rate of QG56 cells was significantly increased after CPP treatment for 48 h (P<0.05). The expression of bax mRNA was increased in QG56 exposed to CPP. The result of immunocytochemis-try indicated that CPP up-regulated the expression of bax protein. Conclusion CPP showed significant inhibitory effect on human lung cancer cell lines QG56 through inducing cell cycle arrest and apoptosis.

RESUMEN

Objective: To study the dynamic variation of Mn, Cu, Zn, Fe, K, Ca, and Mg in velamen and stem bark of Periploca sepium and their correlation with the elements such as periplocin and 4-methoxysalicylic aldehyde in different harvesting periods, and to reveal dominant factors on the accumulation of two active components. Methods: The contents of periplocin and 4-methoxysalicylic aldehyde were detected by FAAS and electric heating wet digestion, and the data were analyzed by SPSS 19.0. Results: The seven elements were varied widely in velamen and stem bark. The variation trends of Ca and periplocin, Fe and 4-methoxysalicylic aldehyde in velamen were similar, equally, those of Fe and 4-methoxysalicylic aldehyde, Ca and Mg in stem bark were similar and like double peaks. Periplocin has highly significant positive correlation with Ca in velamen and significant negative correlation with Zn in stem bark. To 4-methoxysalicylic aldehyde, when in velamen, it has highly significant positive correlation with 4-methoxysalicylic aldehyde and Fe in stem bark, significant negative correlation with Mn in stem bark, and significant positive correlation with Fe in velamen; when in stem bark, it has highly significant positive correlation with Fe in both velamen and stem bark. Conclusion: Ca, Zn, Fe, and Mn are dominant elements of accumulation, the key factors for periplocin are Ca in velamen and Zn in stem bark, and Mg in velamen is indierect acting factors; The key factors for 4-methoxysalicylic aldehyde are Mn in stem bark and Fe, Ca and Mg in stem bark are indirect acting factors.

RESUMEN

OBJECTIVES: The root barks of Periploca sepium Bge. (P. sepium) has been used in traditional Chinese medicine for healing wounds and treating rheumatoid arthritis. However, toxicity in high-doses was often diagnosed by the presence of many glycosides. The potential mutagenicity of P. sepium was investigated both in vitro and in vivo. METHODS: This was examined by the bacterial reverse mutation (Ames) test using Escherichia coli WP2uvrA and Salmonella typhimurium strains, such as TA98, TA100, TA1535, and TA1537. Chromosomal aberrations were investigated using Chinese hamster lung cells, and the micronucleus test using mice. RESULTS: P. sepium did not induce mutagenicity in the bacterial test or chromosomal aberrations in Chinese hamster lung cells, although metabolic activation and micronucleated polychromatic erythrocytes were seen in the mice bone marrow cells. CONCLUSIONS: Considering these results, it is suggested that P. sepium does not have mutagenic potential under the conditions examined in each study.

RESUMEN

OBJECTIVES: The root barks of Periploca sepium Bge. (P. sepium) has been used in traditional Chinese medicine for healing wounds and treating rheumatoid arthritis. However, toxicity in high-doses was often diagnosed by the presence of many glycosides. The potential mutagenicity of P. sepium was investigated both in vitro and in vivo. METHODS: This was examined by the bacterial reverse mutation (Ames) test using Escherichia coli WP2uvrA and Salmonella typhimurium strains, such as TA98, TA100, TA1535, and TA1537. Chromosomal aberrations were investigated using Chinese hamster lung cells, and the micronucleus test using mice. RESULTS: P. sepium did not induce mutagenicity in the bacterial test or chromosomal aberrations in Chinese hamster lung cells, although metabolic activation and micronucleated polychromatic erythrocytes were seen in the mice bone marrow cells. CONCLUSIONS: Considering these results, it is suggested that P. sepium does not have mutagenic potential under the conditions examined in each study.

Asunto(s)

Animales , Cricetinae , Ratones , Artritis Reumatoide , Biotransformación , Médula Ósea , Aberraciones Cromosómicas , Cricetulus , Eritrocitos , Escherichia coli , Glicósidos , Pulmón , Medicina Tradicional China , Pruebas de Micronúcleos , Periploca , Salmonella typhimurium

RESUMEN

Objective To increase the ultimate yield of periplocin in Periploca sepium adventitious root cultures by a two-stage culture based on nitrogen source.Methods Firstly,the effects of nitrogen source(NH-NO-)at different ratios and different total initial nitrogen amounts on the accumulation of biomass and secondary metabolites in adventitious root cultures of P sepium were investigated,and growth and production media for the two-stage culture based on the above results were established.Results The highest biomass and periplocin content were obtained in the culture medium of 15 mmol/L total nitrogen amount with NH-NO(1:2)and 30 mmol/L total nitrogen amount with nitrate as the sole nitrogen source.By adopting a fed-batch cultivation strategy,the dry weight adventitious root,periplocin content and yield were increased by 136%,108%,and 389%,respectively when compared with those of the control,reaching up to 8.13 g/L,157.15 μg/g,and 1277.63 μg/L,respectively.Furthermore,it was found that in the process of two-stage culture,the adventitious roots grew thicker significantly after they were transferred into production medium directly.Conclusion The ultimate yield of periplocin in P.sepium adventitious root cultures could be significantly increased by a two-stage culture based on nitrogen source.