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1.
Molecules ; 27(10)2022 May 20.
Artigo em Inglês | MEDLINE | ID: mdl-35630779

RESUMO

Ranunculus sceleratus L.(RS) has shown various pharmacological effects in traditional Chinese medicine. In our previous study, the positive therapeutic effect on α-naphthylisothiocyanate induced intrahepatic cholestasis in rats was obtained using TianJiu treatment with fresh RS. However, the chemical profile of RS has not been clearly clarified, which impedes the research progress on the therapeutic effect of RS. Herein, an ultra-high performance liquid chromatography coupled with quadrupole Orbitrap high-resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS) method was developed to rapidly separate and identify multiple constituents in the 80% methanol extract of RS. A total of sixty-nine compounds (19 flavonoids, 22 organic acids, 6 coumarins, 4 lignans, 14 nitrogenous compounds, and 4 anthraquinones) were successfully characterized. A total of 12 of these compounds were unambiguously identified by standard samples. Their mass spectrometric fragmentation pathways were investigated. It is worth noting that flavonoids and lignans were identified for the first time in RS. In this study, we successfully provide the first comprehensive report on identifying major chemical constituents in RS by UHPLC-Q-Orbitrap HRMS. The obtained results enrich the RS chemical profile, paving the way for further phytochemical study, quality control, and pharmacological investigation of RS.


Assuntos
Lignanas , Ranunculus , Animais , Cromatografia Líquida de Alta Pressão/métodos , Flavonoides/química , Cromatografia Gasosa-Espectrometria de Massas , Espectrometria de Massas/métodos , Ratos
2.
Pharm Biol ; 60(1): 300-307, 2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-35142600

RESUMO

CONTEXT: Ranunculus ternatus Thunb (Ranunculaceae), (RTT) is used clinically for the treatment of tuberculosis or as tumour adjuvant therapy, but its potential effect on diabetic nephropathy (DN) has not been studied. OBJECTIVE: To investigate the effect of RTT extract in renal fibrosis of DN. MATERIALS AND METHODS: C57BL/6 mice were randomly divided into four groups (n = 12). Diabetes mellitus (DM) mice were induced by streptozotocin (STZ, 55 mg/kg/day) for five consecutive days and treated by RTT extract (2 g/kg). Afterward, blood glucose, HE and Masson staining were assayed. The expression levels of Vimentin, ɑ-SMA, TNF-ɑ, NF-κB p-p65, NF-κB p65, SMYD2, H3K36me3, H3K4me3 were determined by western blots. Firbronectin was respectively assayed by western blot and immunofluorescent staining. RESULTS: RTT extract significantly ameliorated renal injury and renal fibrosis in the renal tissue of STZ-induced diabetic mice as demonstrated by the decreased expression level of Fibronectin (65%), Vimentin and α-SMA (75% & 53%). In addition, the levels of TNF-α (57%), NF-κB p-p65 and NF-κB p65 (35% & 25%) were elevated in the DN mice. Importantly, these were alleviated after RTT extract treatment. Moreover, we observed that the protein levels of SMYD2 (30%), H3K36me3 and H3K4me3 (53% & 75%) were reduced in DN mice after treatment with RTT extract. DISCUSSION AND CONCLUSIONS: RTT extract mediates antifibrotic effects and anti-inflammatory responses in STZ-induced DN mainly through suppressing SMYD2 activation and H3K36me3 and H3K4me3 protein expression. RTT extract might have therapeutic potential against high glucose-induced nephropathy.


Assuntos
Diabetes Mellitus Experimental/tratamento farmacológico , Nefropatias Diabéticas/tratamento farmacológico , Extratos Vegetais/farmacologia , Ranunculus/química , Animais , Anti-Inflamatórios/isolamento & purificação , Anti-Inflamatórios/farmacologia , Glicemia/efeitos dos fármacos , Fibrose , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Estreptozocina
3.
J Med Virol ; 94(6): 2727-2735, 2022 06.
Artigo em Inglês | MEDLINE | ID: mdl-35075662

RESUMO

The chronic hepatitis B virus (HBV) infection is a worldwide public health problem, which cannot be cured by current therapeutics due to the persistence of viral CCC DNA in the infected hepatocytes. Screening from medicinal herbs for anti-HBV activities showed that the ethanol extract from Ranunculus japonicus Thunb. could decrease the production of HBV e antigen (HBeAg). Further study showed that the extract had no effect on core protein expression but significantly reduced the efficiency of viral capsid assembly. The levels of viral pgRNA and total core DNA were not affected significantly. However, the ratio of RC DNA/SS DNA decreased, indicating that the conversion of RC DNA from SS DNA was delayed by the extract. More interestingly, though similar levels of RC DNA were accumulated, the CCC DNA level and its formation efficiency were reduced significantly, which was also consistent with the decreased level of HBeAg, indicating that R. japonicus Thunb. extract could inhibit the CCC DNA formation. Together, this study found that R. japonicus Thunb. extract could inhibit HBV replication at multiple steps, especially showed significant inhibitory effects on capsid assembly and CCC DNA formation.


Assuntos
Hepatite B Crônica , Hepatite B , Ranunculus , DNA Circular , DNA Viral/genética , Etanol/metabolismo , Etanol/farmacologia , Antígenos E da Hepatite B/metabolismo , Vírus da Hepatite B/genética , Humanos , Extratos Vegetais/farmacologia , Ranunculus/genética , Ranunculus/metabolismo , Replicação Viral
4.
Nat Prod Res ; 36(4): 961-966, 2022 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33280438

RESUMO

Ranunculus muricatus L., an important member of family Ranunculaceae upon submission to phytochemical studies, led to the isolation of a novel natural hydrazine derivative, muricazine (1). Chemical structure of the compound was established with the aid of advanced spectroscopic techniques. It was evaluated for in vitro antioxidant, lipoxygenase, and urease (jack-bean) inhibitory activities. Results suggested that compound 1 could scavenge the DPPH free radical (42.1 ± 0.12 µM) to a great extent as compared to the standard (40.6 ± 0.91 µM). However, it showed moderate inhibitory potential against lipoxygenase (65.2 ± 0.45 µM) and urease (54.8 ± 0.23 µM) enzymes.


Assuntos
Inibidores Enzimáticos , Hidrazinas , Ranunculus , Antioxidantes , Inibidores Enzimáticos/química , Inibidores Enzimáticos/farmacologia , Hidrazinas/química , Hidrazinas/farmacologia , Lipoxigenase , Inibidores de Lipoxigenase , Ranunculus/química , Urease/antagonistas & inibidores
5.
Lett Appl Microbiol ; 74(1): 84-91, 2022 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-34637547

RESUMO

Ranunculus (Ranunculus asiaticus L.) is one of the most popular cut flowers in Japan. However, the infection rate of ranunculus mild mosaic virus (RanMMV) in ranunculus plants has been gradually increasing during cultivation, suggesting that RanMMV may be transmitted from weeds to ranunculus plants in cultivation fields. In our survey, RanMMV in R. japonicus, R. tachiroei, R. cantoniensis, Geranium carolinianum, Vicia sativa, V. tetrasperma and V. hirsute in ranunculus fields and noncultivation regions in Japan was detected. Ranunculaceae weeds grow all year in cultivation fields, unlike R. asiaticus, indicating that these weeds may be a source of RanMMV infection. In addition, a pairwise comparison of CP genes between RanMMV isolates taken from R. asiaticus, R. japonicus, and R. tachiroei showed high nucleotide (98·1-100%) and amino acid (98·5-100%) identities. These results support the hypothesis that RanMMV may be transmitted between Ranunculaceae weeds and R. asiaticus plants. Thus, virus control should focus on removing host weeds from the cultivation fields.


Assuntos
Vírus do Mosaico , Ranunculus , Flores , Japão , Ranunculus/genética
6.
Molecules ; 26(24)2021 Dec 10.
Artigo em Inglês | MEDLINE | ID: mdl-34946555

RESUMO

Leishmaniasis and schistosomiasis are neglected tropical diseases (NTDs) infecting the world's poorest populations. Effectiveness of the current antileishmanial and antischistosomal therapies are significantly declining, which calls for an urgent need of new effective and safe drugs. In Ethiopia fresh leaves of Ranunculus multifidus Forsk. are traditionally used for the treatment of various ailments including leishmaniasis and eradication of intestinal worms. In the current study, anemonin isolated from the fresh leaves of R. multifidus was assessed for its in vitro antileishmanial and antischistosomal activities. Anemonin was isolated from the hydro-distilled extract of the leaves of R. multifidus. Antileishmanial activity was assessed on clinical isolates of the promastigote and amastigote forms of Leishmania aethiopica and L. donovani clinical isolates. Resazurin reduction assay was used to determine antipromastigote activity, while macrophages were employed for antiamastigote and cytotoxicity assays. Antischistosomal assays were performed against adult Schistosoma mansoni and newly transformed schistosomules (NTS). Anemonin displayed significant antileishmanial activity with IC50 values of 1.33 nM and 1.58 nM against promastigotes and 1.24 nM and 1.91 nM against amastigotes of L. aethiopica and L. donovani, respectively. It also showed moderate activity against adult S. mansoni and NTS (49% activity against adult S. mansoni at 10 µM and 41% activity against NTS at 1 µM). The results obtained in this investigation indicate that anemonin has the potential to be used as a template for designing novel antileishmanial and antischistosomal pharmacophores.


Assuntos
Antiprotozoários/farmacologia , Furanos/farmacologia , Leishmania/efeitos dos fármacos , Extratos Vegetais/farmacologia , Ranunculus/química , Schistosoma mansoni/efeitos dos fármacos , Animais , Antiprotozoários/química , Antiprotozoários/isolamento & purificação , Furanos/química , Furanos/isolamento & purificação , Testes de Sensibilidade Parasitária , Extratos Vegetais/química , Extratos Vegetais/isolamento & purificação , Folhas de Planta/química
7.
Genes (Basel) ; 12(11)2021 11 18.
Artigo em Inglês | MEDLINE | ID: mdl-34828424

RESUMO

Alpine habitats are shaped by harsh abiotic conditions and cold climates. Temperature stress can affect phenotypic plasticity, reproduction, and epigenetic profiles, which may affect acclimation and adaptation. Distribution patterns suggest that polyploidy seems to be advantageous under cold conditions. Nevertheless, whether temperature stress can induce gene expression changes in different cytotypes, and how the response is initialized through gene set pathways and epigenetic control remain vague for non-model plants. The perennial alpine plant Ranunculus kuepferi was used to investigate the effect of cold stress on gene expression profiles. Diploid and autotetraploid individuals were exposed to cold and warm conditions in climate growth chambers and analyzed via transcriptome sequencing and qRT-PCR. Overall, cold stress changed gene expression profiles of both cytotypes and induced cold acclimation. Diploids changed more gene set pathways than tetraploids, and suppressed pathways involved in ion/cation homeostasis. Tetraploids mostly activated gene set pathways related to cell wall and plasma membrane. An epigenetic background for gene regulation in response to temperature conditions is indicated. Results suggest that perennial alpine plants can respond to temperature extremes via altered gene expression. Tetraploids are better acclimated to cold conditions, enabling them to colonize colder climatic areas in the Alps.


Assuntos
Aclimatação , Resposta ao Choque Frio , Poliploidia , Ranunculus/genética , Transcriptoma , Epigênese Genética , Regulação da Expressão Gênica de Plantas , Genótipo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Ranunculus/metabolismo
8.
Molecules ; 26(20)2021 Oct 13.
Artigo em Inglês | MEDLINE | ID: mdl-34684762

RESUMO

The leaves of Ranunculus multifidus Forsk. are traditionally used for the treatment of malaria in several African countries. In the present study, 80% methanol (RM-M) and hydrodistilled (RM-H) extracts of fresh leaves from R. multifidus and its major constituent anemonin were tested for their in vivo antimalarial activity against Plasmodium berghei in mice. Anemonin was also tested for its in vitro antimycobacterial activity against Mycobacterium smegmatis and M. abscessus in a microbroth dilution assay, and bacterial growth was analyzed by OD measurement. The isolation of anemonin from RM-H was carried out using preparative thin layer chromatography (PTLC). The chemical structures of anemonin and its hydrolysis product were elucidated using spectroscopic methods (HR-MS; 1D and 2D-NMR). Results of the study revealed that both RM-M and RM-H were active against P. berghei in mice, although the latter demonstrated superior activity (p < 0.001), as compared to the former. At a dose of 35.00 mg/kg/day, RM-H demonstrated a chemosuppression value of 70% in a 4-day suppressive test. In a 4-day suppressive, Rane's and prophylactic antimalarial tests, anemonin showed median effective doses (ED50s) of 2.17, 2.78 and 2.70 µM, respectively. However, anemonin did not inhibit the growth of M. smegmatis and M. abscessus.


Assuntos
Antimaláricos/farmacologia , Furanos/farmacologia , Ranunculus/metabolismo , Animais , Antimaláricos/metabolismo , Modelos Animais de Doenças , Etiópia , Feminino , Furanos/química , Malária/tratamento farmacológico , Masculino , Camundongos , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Folhas de Planta/metabolismo , Plasmodium berghei/efeitos dos fármacos , Plasmodium falciparum/efeitos dos fármacos
9.
Mol Ecol ; 30(11): 2659-2675, 2021 06.
Artigo em Inglês | MEDLINE | ID: mdl-33871107

RESUMO

Geographical parthenogenesis (GP) describes the phenomenon that apomicts tend to have larger distribution areas and/or occur at higher altitudes or latitudes compared to sexual relatives. However, the complex effects of genome-wide heterozygosity, ploidy, reproduction mode (sexual vs. apomictic), and environment shaping GP of plants are still not well understood. We ascertained ploidy and reproduction mode by flow cytometry of 221 populations, and added genomic RADseq data (maximum 33,165 loci) of 80 taxa of the Ranunculus auricomus polyploid plant complex in temperate Europe. We observed 7% mainly diploid sexual, 28% facultative apomictic (mean sexuality 7.1%), and 65% obligate apomictic populations. Sexuals occupied a more southern, smaller distribution area, whereas apomicts expanded their range to higher latitudes. Within the complex, we detected three main genetic clusters and highly reticulate relationships. A genetically-informed path analysis using GLMMs revealed several significant relationships. Sexuality of populations (percent of sexual seeds) was higher in diploids compared to polyploids, associated with more petals, and similar between forests and open habitats. In contrast to other apomictic plant complexes, sexuality was mainly positively correlated to solar radiation and isothermality, which fits the southern distribution. We found up to three times higher heterozygosity in polyploids compared to diploids, and generally more heterozygous individuals in forests compared with open habitats. Interestingly, we revealed a previously unknown positive association between heterozygosity and temperature seasonality, suggesting a higher resistance of polyploids to more extreme climatic conditions. We provide empirical evidence for intrinsic and extrinsic factors shaping the GP pattern in a polyploid plant complex.


Assuntos
Poliploidia , Ranunculus , Diploide , Europa (Continente) , Humanos , Partenogênese/genética
10.
J Ethnopharmacol ; 271: 113818, 2021 May 10.
Artigo em Inglês | MEDLINE | ID: mdl-33465444

RESUMO

ETHNOPHARMACOLOGICAL RELEVANCE: Ranunculus japonicus Thunb. (short for R. japonicus) is a topically applied herb with the activities of removing jaundice, nebula and edema, preventing malaria, stopping asthma, promoting diuresis and relieving pain. It was firstly recorded in Zhouhou Beiji Fang and has been used for the treatment of malaria, ulcers, carbuncle, jaundice, migraine, stomachache, toothache and arthritis for over 1800 years. AIM OF THE STUDY: This study aimed to uncover the potentially effective components of R. japonicus and the pharmacological mechanisms against rheumatoid arthritis (RA) by combing LC-MS and network pharmacology. MATERIALS AND METHODS: Firstly, the chemical constituents of R. japonicus were qualitatively identified by UPLC-ESI-LTQ-Orbitrap MS. Then we performed target prediction by PharmMapper, protein-protein interaction (PPI) analysis via String, GO and KEGG pathway enrichment analysis by DAVID and constructed the compound-target-pathway network using Cytoscape. Thirdly, crucial compounds in the network were quantitatively analyzed to achieve quality control of R. japonicus. Finally, the pharmacological activities of R. japonicus and two potentially bioactive ingredients were validated in RA-FLSs (Rheumatoid Arthritis Fibroblast-like Synoviocytes) in vitro. RESULTS: Overall fifty-four ingredients of R. japonicus were identified and forty-five components were firstly discovered in R. japonicus. Among them, twenty-seven validated compounds were predicted to act on twenty-five RA-related targets and they might exhibit therapeutic effects against RA via positive regulation of cell migration, etc. Nine potentially bioactive components of R. japonicus which played important roles in the compound-target-pathway network were simultaneously quantified by an optimized UPLC-ESI-Triple Quad method. In vitro, compared to control group, R. japonicus extract, berberine and yangonin significantly inhibited the migration capacity of RA-FLSs after 24 h treatment. CONCLUSION: This study clarified that R. japonicus and the bioactive ingredients berberine and yangonin might exert therapeutic actions for RA via suppressing the aggressive phenotypes of RA-FLSs through combined LC-MS technology and network pharmacology tools for the first time. The present research provided deeper understanding into the chemical profiling, pharmacological activities and quality control of R. japonicus and offered reference for further scientific research and clinical use of R. japonicus in treating RA.


Assuntos
Artrite Reumatoide/tratamento farmacológico , Medicamentos de Ervas Chinesas/química , Medicamentos de Ervas Chinesas/farmacologia , Farmacologia/métodos , Ranunculus/química , Movimento Celular/efeitos dos fármacos , Células Cultivadas , Cromatografia Líquida , Fibroblastos/efeitos dos fármacos , Humanos , Compostos Fitoquímicos/química , Compostos Fitoquímicos/farmacologia , Mapas de Interação de Proteínas/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Sinoviócitos/efeitos dos fármacos , Espectrometria de Massas em Tandem , Cicatrização/efeitos dos fármacos
11.
J Ethnopharmacol ; 265: 113347, 2021 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-32890715

RESUMO

ETHNOPHARMACOLOGICAL RELEVANCE: The members of the genus Ranunculus have counter-irritating properties and thus, they are traditionally used for treating anti-inflammatory disorders and other skin conditions. Ranunculus macrophyllus Desf. is a wild medicinal plant growing in Algeria and traditionally used to treat some cutaneous skin disorders. AIM: The aim of this study was to characterize the composition of the ethyl acetate and n-butanol extracts from Ranunculus macrophyllus Desf. as well as to elucidate and to compare their effect against acute skin inflammation. Moreover, both the antioxidant activity and the acute toxicity of the plant extracts were also studied. MATERIALS AND METHODS: Spectrophotometric and chromatographic methods were employed to identify and quantify phenolic compounds and triterpenoids from R. macrophyllus Desf. fractions. The antioxidant activity was estimated using the phosphomolebdenum, DPPH, reducing power and ß-carotene bleaching assays. The ethyl acetate and n-butanol extracts were screened for their anti-inflammatory activities using ex-vivo membrane stabilizing assays and in-vivo acute skin inflammation model. RESULTS: Ethyl acetate fraction showed the highest amounts of total phenolic compounds (413 ± 4 µg GAE/mg extract) and triterpenoids (70.4 ± 1.8 µg UAE/mg extract). Rutin, hesperidin, myricetin and kaempferol were the major compounds identified in the different fractions. Ethyl acetate fraction exhibited strong DPPH• radical scavenging ability (IC50 1.6 ± 0.2 µg/mL), high total antioxidant capacity (447 ± 7 µg AAE/mg extract) and reducing power (514 ± 8 µg AAE/mg extract). Ethyl acetate fraction inhibited (73.4 ± 0.3) % of linoleic acid peroxidation. Ethyl acetate and n-butanol fractions did not have any visible toxicity at 2000 mg/kg and presented excellent membrane stabilizing ability. The inhibition of xylene induced ear inflammation was (38 ± 4) % and (46 ± 1) % for RM-B and RM-EA, respectively. CONCLUSIONS: The high content of both phenolic compounds and triterpenoids combined with the remarkable anti-inflammatory effect and antioxidant activity of ethyl acetate and n-butanol extracts from R. macrophyllus Desf. support the wide spread use of this traditional plant on some skin disorders (inflammatory skin disorders).


Assuntos
Anti-Inflamatórios/farmacologia , Fenóis/farmacologia , Extratos Vegetais/farmacologia , Ranunculus/química , 1-Butanol/química , Acetatos/química , Argélia , Animais , Anti-Inflamatórios/isolamento & purificação , Antioxidantes/isolamento & purificação , Antioxidantes/farmacologia , Modelos Animais de Doenças , Feminino , Inflamação/tratamento farmacológico , Inflamação/patologia , Masculino , Camundongos , Fenóis/isolamento & purificação , Extratos Vegetais/química
12.
Nat Prod Res ; 35(23): 4994-5000, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-32352323

RESUMO

Muriolide (1), a new aromatic lactone, has been isolated from the ethyl acetate fraction of Ranunculus muricatus. The compound was structurally characterized with the help of UV, IR, mass, 1D- and 2D-NMR data. It was tested in vitro for antioxidant and lipoxygenase inhibitory potential. Compound 1 showed good DPPH radical scavenging activity (IC50=56.9 µM), however it was moderately active against lipoxygenase enzyme (IC50=68.3 µM).


Assuntos
Ranunculus , Antioxidantes , Lactonas , Lipoxigenase , Extratos Vegetais
13.
Int J Mol Sci ; 21(22)2020 Nov 19.
Artigo em Inglês | MEDLINE | ID: mdl-33228190

RESUMO

Low temperature stress has a severe impact on the distribution, physiology, and survival of plants in their natural habitats. While numerous studies have focused on the physiological and molecular adjustments to low temperatures, this study provides evidence that cold induced physiological responses coincide with distinct ultrastructural alterations. Three plants from different evolutionary levels and habitats were investigated: The freshwater alga Micrasterias denticulata, the aquatic plant Lemna sp., and the nival plant Ranunculus glacialis. Ultrastructural alterations during low temperature stress were determined by the employment of 2-D transmission electron microscopy and 3-D reconstructions from focused ion beam-scanning electron microscopic series. With decreasing temperatures, increasing numbers of organelle contacts and particularly the fusion of mitochondria to 3-dimensional networks were observed. We assume that the increase or at least maintenance of respiration during low temperature stress is likely to be based on these mitochondrial interconnections. Moreover, it is shown that autophagy and degeneration processes accompany freezing stress in Lemna and R. glacialis. This might be an essential mechanism to recycle damaged cytoplasmic constituents to maintain the cellular metabolism during freezing stress.


Assuntos
Araceae/fisiologia , Autofagia/fisiologia , Cloroplastos/fisiologia , Micrasterias/fisiologia , Mitocôndrias/fisiologia , Ranunculus/fisiologia , Organismos Aquáticos , Araceae/ultraestrutura , Respiração Celular/fisiologia , Cloroplastos/ultraestrutura , Temperatura Baixa , Resposta ao Choque Frio , Retículo Endoplasmático/fisiologia , Retículo Endoplasmático/ultraestrutura , Micrasterias/ultraestrutura , Microscopia Eletrônica de Transmissão , Mitocôndrias/ultraestrutura , Peroxissomos/fisiologia , Peroxissomos/ultraestrutura , Fotossíntese/fisiologia , Células Vegetais/fisiologia , Células Vegetais/ultraestrutura , Ranunculus/ultraestrutura
14.
Biomolecules ; 10(11)2020 11 17.
Artigo em Inglês | MEDLINE | ID: mdl-33212893

RESUMO

Ranunculus muricatus L. is a spiny fruit buttercup that is used in various traditional medicinal systems. In the current investigation of R. muricatus, the new chalcone 4-benzyloxylonchocarpin (1), the new anthraquinone muracatanes A (2), the new-to-nature anthraquinone muracatanes B (3), and the new naphthalene analog muracatanes C (4) were isolated, in addition to the three previously reported compounds, 4-methoxylonchocarpin (5), ß-sitosterol (6), and ß-sitosterol ß-D-glucopyranoside (7). Their structures were elucidated using 1D (1H and 13C) and 2D (COSY, HSQC, and HMBC) NMR spectroscopy and HR-ESI-MS. Chalcone 1 showed potent acetylcholinesterase inhibitory effects with Ki of 5.39 µM and Ki' of 3.54 µM, but none of the isolated compounds showed inhibitory activity towards butyrylcholinesterase. Anthraquinone 3 illustrated α-glucosidase inhibitory effects with IC50-values of 164.46 ± 83.04 µM. Compound 5 displayed moderate cytotoxic activity towards ovarian carcinoma (A2780, IC50 = 25.4 µM), colorectal adenocarcinoma (HT29, IC50 = 20.2 µM), breast cancer (MCF7, IC50 = 23.7 µM), and thyroid carcinoma (SW1736, IC50 = 26.2 µM) while it was inactive towards pharynx carcinoma (FaDu: IC50 > 30 µM).


Assuntos
Inibidores da Colinesterase/farmacologia , Inibidores de Glicosídeo Hidrolases/farmacologia , Simulação de Acoplamento Molecular/métodos , Extratos Vegetais/farmacologia , Ranunculus , Animais , Inibidores da Colinesterase/química , Inibidores da Colinesterase/isolamento & purificação , Electrophorus , Inibidores de Glicosídeo Hidrolases/química , Inibidores de Glicosídeo Hidrolases/isolamento & purificação , Cavalos , Extratos Vegetais/química , Extratos Vegetais/isolamento & purificação
15.
BMC Plant Biol ; 20(1): 523, 2020 Nov 17.
Artigo em Inglês | MEDLINE | ID: mdl-33203395

RESUMO

BACKGROUND: Hybridization and polyploidization are powerful evolutionary factors that are associated with manifold developmental changes in plants such as irregular progression of meiosis and sporogenesis. The emergence of apomixis, which is asexual reproduction via seeds, is supposed to be connected to these factors and was often regarded as an escape from hybrid sterility. However, the functional trigger of apomixis is still unclear. Recently formed di- and polyploid Ranunculus hybrids, as well as their parental species were analysed for their modes of mega- and microsporogenesis by microscopy. Chromosomal configurations during male meiosis were screened for abnormalities. Meiotic and developmental abnormalities were documented qualitatively and collected quantitatively for statistical evaluations. RESULTS: Allopolyploids showed significantly higher frequencies of erroneous microsporogenesis than homoploid hybrid plants. Among diploids, F2 hybrids had significantly more disturbed meiosis than F1 hybrids and parental plants. Chromosomal aberrations included laggard chromosomes, chromatin bridges and disoriented spindle activities. Failure of megasporogenesis appeared to be much more frequent in than of microsporogenesis is correlated to apomixis onset. CONCLUSIONS: Results suggest diverging selective pressures on female and male sporogenesis, with only minor effects of hybridity on microsporogenesis, but fatal effects on the course of megasporogenesis. Hence, pollen development continues without major alterations, while selection will favour apomixis as alternative to the female meiotic pathway. Relation of investigated errors of megasporogenesis with the observed occurrence of apospory in Ranunculus hybrids identifies disturbed female meiosis as potential elicitor of apomixis in order to rescue these plants from hybrid sterility. Male meiotic disturbance appears to be stronger in neopolyploids than in homoploid hybrids, while disturbances of megasporogenesis were not ploidy-dependent.


Assuntos
Apomixia , Gametogênese Vegetal , Poliploidia , Ranunculus/fisiologia , Evolução Biológica , Diploide , Hibridização Genética , Meiose , Ranunculus/genética , Sementes/genética , Sementes/fisiologia
16.
PLoS One ; 15(10): e0240121, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33017445

RESUMO

Worldwide, the genus Ranunculus includes approximately 600 species and is highly genetically diverse. Recent taxonomic reports suggest that the genus has a monophyletic origin, divided into two subgenera, and consists of 17 sections. The Central Asian country of Kazakhstan has 62 species of the genus that have primarily been collected in the central part of the country. The latest collection trips in southern parts of the country have led to the description of a wider distribution area for Ranunculus and the identification of a new species Ranunculus talassicus Schegol. et A.L. Ebel from Western Tien Shan. Therefore, in this study, attempts were made to assess the molecular taxonomic positions of R. talassicus and two other species endemic to the Central Asian region R. karkaralensis Schegol. and R. pskemensis V.N. Pavlov in relation to other species of the genus, using internal transcribed spacer (ITS) molecular genetic markers. The ITS-aligned sequences of 22 local Central Asian accessions and 43 accession sequences available in the National Center for Biotechnology Information (NCBI) database allowed the construction of a maximum parsimony phylogenetic tree and a Neighbor-Net network. The results indicated that R. talassicus and R. pskemensis could be assigned to section Ranunculastrum. Additionally, an assessment of the network suggested that R. pskemensis was the rooting taxon for the group of species containing R. talassicus, and that R. illyricus L. and R. pedatus Waldst. & Kit. were founders of a prime rooting node for the Ranunculastrum section of the genus. The ITS-aligned sequences showed that R. karkaralensis was indifferent with respect to three other species in the Ranunculus section of the genus, i.e., R. acris L., R. grandifolius C.A. Mey., and R. subborealis Tzvelev. The study indicated that the assessments of ITS-based phylogenetic tree and Neighbor-Net network provided new insights into the taxonomic positions of three endemic species from Central Asia.


Assuntos
DNA de Plantas/genética , Ranunculus/genética , Ásia Central , Variação Genética , Filogenia , Ranunculus/classificação
17.
Int J Mol Sci ; 21(19)2020 Sep 24.
Artigo em Inglês | MEDLINE | ID: mdl-32987913

RESUMO

Ranunculus glacialis grows and reproduces successfully, although the snow-free time period is short (2-3 months) and night frosts are frequent. At a nival site (3185 m a.s.l.), we disentangled the interplay between the atmospheric temperature, leaf temperatures, and leaf freezing frequency to assess the actual strain. For a comprehensive understanding, the freezing behavior from the whole plant to the leaf and cellular level and its physiological after-effects as well as cell wall chemistry were studied. The atmospheric temperatures did not mirror the leaf temperatures, which could be 9.3 °C lower. Leaf freezing occurred even when the air temperature was above 0 °C. Ice nucleation at on average -2.6 °C started usually independently in each leaf, as the shoot is deep-seated in unfrozen soil. All the mesophyll cells were subjected to freezing cytorrhysis. Huge ice masses formed in the intercellular spaces of the spongy parenchyma. After thawing, photosynthesis was unaffected regardless of whether ice had formed. The cell walls were pectin-rich and triglycerides occurred, particularly in the spongy parenchyma. At high elevations, atmospheric temperatures fail to predict plant freezing. Shoot burial prevents ice spreading, specific tissue architecture enables ice management, and the flexibility of cell walls allows recurrent freezing cytorrhysis. The peculiar patterning of triglycerides close to ice rewards further investigation.


Assuntos
Parede Celular/fisiologia , Resposta ao Choque Frio , Células do Mesofilo , Ranunculus/fisiologia , Congelamento , Gelo , Células do Mesofilo/citologia , Células do Mesofilo/fisiologia , Fotossíntese
18.
Genes (Basel) ; 11(7)2020 06 30.
Artigo em Inglês | MEDLINE | ID: mdl-32630035

RESUMO

Apomixis, the asexual reproduction via seeds, is associated to polyploidy and hybridization. To identify possible signatures of apomixis, and possible candidate genes underlying the shift from sex to apomixis, microarray-based gene expression patterns of live microdissected ovules at four different developmental stages were compared between apomictic and sexual individuals of the Ranunculus auricomus complex. Following predictions from previous work on mechanisms underlying apomixis penetrance and expressivity in the genus, gene expression patterns were classified into three categories based on their relative expression in apomicts compared to their sexual parental ancestors. We found evidence of misregulation and differential gene expression between apomicts and sexuals, with the highest number of differences detected during meiosis progression and emergence of aposporous initial (AI) cells, a key developmental stage in the ovule of apomicts where a decision between divergent reproductive pathways takes place. While most of the differentially expressed genes (DEGs) could not be annotated, gene expression was classified into transgressive, parent of origin and ploidy effects. Genes related to gametogenesis and meiosis demonstrated patterns reflective of transgressive and genome dosage effects, which support the hypothesis of a dominant factor controlling apomixis in Ranunculus and modulated by secondary modifiers. Three genes with probable functions in sporogenesis and gametogenesis development are identified and characterized for future studies.


Assuntos
Apomixia , Genes de Plantas , Óvulo Vegetal/genética , Ranunculus/genética , Gametogênese , Regulação da Expressão Gênica de Plantas , Óvulo Vegetal/metabolismo , Ranunculus/fisiologia
19.
Mol Ecol ; 29(11): 2031-2049, 2020 06.
Artigo em Inglês | MEDLINE | ID: mdl-32374933

RESUMO

The time frame and geographical patterns of diversification processes in European temperate-montane herbs are still not well understood. We used the sexual species of the Ranunculus auricomus complex as a model system to understand how vicariance versus dispersal processes in the context of Pleistocene climatic fluctuations have triggered speciation in temperate-montane plant species. We used target enrichment sequence data from about 600 nuclear genes and coalescent-based species tree inference methods to resolve phylogenetic relationships among the sexual taxa of the complex. We estimated absolute divergence times and, using ancestral range reconstruction, we tested if speciation was enhanced by vicariance or by dispersal processes. Phylogenetic relationships among taxa were fully resolved with some incongruence in the position of the tetraploid R. marsicus. Speciation events took place in a very short time at the end of the Mid-Pleistocene Transition (830-580 thousand years ago [ka]). A second wave of intraspecific geographical differentiation occurred at the end of the Riss glaciation or during the Eemian interglacial between 200 and 100 ka. Ancestral range reconstruction suggests a widespread European ancestor of the R. auricomus complex. Vicariance has triggered allopatric speciation in temperate-montane plant species during the climatic deterioration that occurred during the last phase of the Mid-Pleistocene Transition. Vegetation restructuring from forest into tundra could have confined these forest species into isolated glacial macro- and microrefugia. During subsequent warming periods, range expansions of these species could have been hampered by apomictic derivatives and by other congeneric competitors in the same habitat.


Assuntos
Especiação Genética , Filogenia , Ranunculus , Ecossistema , Europa (Continente) , Florestas , Geografia , Ranunculus/classificação , Ranunculus/genética
20.
Int J Mol Sci ; 21(9)2020 May 07.
Artigo em Inglês | MEDLINE | ID: mdl-32392879

RESUMO

Polyploidization and the shift to apomictic reproduction are connected to changes in DNA cytosine-methylation. Cytosine-methylation is further sensitive to environmental conditions. We, therefore, hypothesize that DNA methylation patterns would differentiate within species with geographical parthenogenesis, i.e., when diploid sexual and polyploid apomictic populations exhibit different spatial distributions. On natural populations of the alpine plant Ranunculus kuepferi, we tested differences in methylation patterns across two cytotypes (diploid, tetraploid) and three reproduction modes (sexual, mixed, apomictic), and their correlation to environmental data and geographical distributions. We used methylation-sensitive amplified fragment-length polymorphism (methylation-sensitive AFLPs) and scored three types of epiloci. Methylation patterns differed independently between cytotypes versus modes of reproduction and separated three distinct combined groups (2x sexual + mixed, 4x mixed, and 4x apomictic), with differentiation of 4x apomicts in all epiloci. We found no global spatial autocorrelation, but instead correlations to elevation and temperature gradients in 22 and 36 epiloci, respectively. Results suggest that methylation patterns in R. kuepferi were altered by cold conditions during postglacial recolonization of the Alps, and by the concomitant shift to facultative apomixis, and by polyploidization. Obligate apomictic tetraploids at the highest elevations established a distinct methylation profile. Methylation patterns reflect an ecological gradient rather than the geographical differentiation.


Assuntos
Análise do Polimorfismo de Comprimento de Fragmentos Amplificados/métodos , Metilação de DNA , DNA de Plantas/genética , Ranunculus/fisiologia , Temperatura Baixa , Diploide , Epigênese Genética , Geografia , Partenogênese , Estresse Fisiológico , Tetraploidia
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