Long-term organogenic callus cultivation of Ranunculus illyricus L.: a blueprint for sustainable ex situ conservation of the species in urban greenery.

The growing trend of introducing wild plant species into urban environments necessitates the identification of novel species adapted to prevailing conditions. A promising reservoir of such species may be xerothermic communities where Ranunculus illyricus occurs. This study aimed to establish a micropropagation protocol for R. illyricus using indirect organogenesis. The protocol includes initiation of culture from various explants, callus proliferation, shoot regeneration, multiplication, and concurrent rooting. Callus appeared on most types of vegetative explants tested, but stolons were considered the best due to their good availability, high disinfection (85%), and robust callus production (maximum increase - 363.1%). The growth rate of the callus fresh matter (CFM) obtained from stolons was calculated. Greater CFM was obtained on the medium with the supplemented picloram 8.0 mg L- 1 with kinetin 5.0 mg L- 1 and in second part of experiment on medium with the addition of 2,4-D (2,4-dichlorophenoxyacetic acid) 2.0 mg L- 1 alone or picloram 6.0 mg L- 1 with kinetin 8.0 mg L- 1. Shoot organogenesis was observed on macronutrients B5 (Gamborg medium), micronutrients MS (Murashige and Skoog) medium with the addition of 2.0 mg L- 1 IBA (indole-3-butyric acid) and 4.0 mg L- 1 BAP (6-benzylaminopurine). To document the process of callus differentiation, microscopic preparations were prepared. Subsequently, the regenerated plants underwent acclimatisation and their growth in an ex situ collection was monitored over three growing seasons. In particular, in vitro-origin plants exhibited developmental patterns similar to those of their seed-origin counterparts. The incorporation of R. illyricus into urban landscapes not only increases aesthetic appeal, but also ensures the preservation of valuable genetic resources for this rare species, potentially contributing to effective ex situ conservation in the future. This marks the first scientific report on in vitro cultures of R. illyricus.

Network pharmacology and transcriptomic profiling elucidate the therapeutic effects of Ranunculus ternatus Thunb on liver fibrosis via MK3-NF-κB inhibition.

Activation of hepatic stellate cells (HSCs) is critical in the progression of liver fibrosis and is a promising target for anti-hepatic fibrosis drug development. Moreover, effective pharmacological interventions targeting this pathomechanism are scarce. Our study confirms the therapeutic value of ß-sitosterol, a major constituent of Ranunculus ternatus Thunb, in hepatic fibrosis and identifies its underlying mechanisms. After treatment with ß-sitosterol, CCL4-induced hepatic fibrosis was reversed in mice, while inflammatory and hepatic fibrosis indices were improved. Meanwhile, we explored the molecular mechanism of ß-sitosterol treatment for hepatic fibrosis and, based on RNA-seq results, found that the ameliorative effect of ß-sitosterol on hepatic fibrosis was associated with the MK3 and NF-κB signalling pathways. MK3, an important kinase in the MAPK pathway, plays a role in transmitting upstream and downstream signals, whereas the NF-κB signalling pathway has been shown to be associated with HSC activation. We verified the interaction between MK3 and IκB in HSC cells using endogenous Co-IP, whereas ß-sitosterol reduced the binding of MK3 to IκB and the activation of the NF-κB signalling pathway. Our findings reveal the mechanism of ß-sitosterol in the treatment of liver fibrosis, suggesting that ß-sitosterol may be a promising drug for the treatment of liver fibrosis and deserves further investigation.

Different intensities and directions of hyporheic water exchange in habitats of aquatic Ranunculus species in rivers-a case study in Poland.

Hyporheic water exchange driven by groundwater-surface water interactions constitutes habitat conditions for aquatic biota. In our study, we conducted a field-research-based analysis of hyporheic water exchange to reveal whether the hyporheic water exchange differentiates particular Ranunculus sp. habitats. We measured the density of the stream of upwelling and hydraulic gradients of water residing in the hyporheic zone in 19 Polish rivers. We revealed that R. peltatus and R. penicillatus persist in habitats of considerably higher hyporheic water exchange upwelling flux (respectively 0.0852 m3∙d-1∙m-2 and 0.0952 m3∙d-1∙m-2) than R. circinatus, R. fluitans, and a hybrid of R. circinatus × R. fluitans (respectively m3∙d-1∙m-2; 0.0222 m3∙d-1∙m-2 and 0.0717 m3∙d-1∙m-2). The presented results can be used to indicate aquatic habitat suitability in the case of protection and management of ecosystems settled by Ranunculus sp.

In vitro and in vivo antitrypanosomal activity of the fresh leaves of Ranunculus Multifidus Forsk and its major compound anemonin against Trypanosoma congolense field isolate.

BACKGROUND: Animal trypanosomiasis is a major livestock problem due to its socioeconomic impacts in tropical countries. Currently used trypanocides are toxic, expensive, and the parasites have developed resistance to the existing drugs, which calls for an urgent need of new effective and safe chemotherapeutic agents from alternative sources such as medicinal plants. In Ethiopian traditional medicine fresh leaves of Ranunculus multifidus Forsk, are used for the treatment of animal trypanosomiasis. The present study aimed to evaluate the antitrypanosomal activity of the fresh leaves of R. multifidus and its major compound anemonin against Trypanosoma congolense field isolate. METHODS: Fresh leaves of R. multifidus were extracted by maceration with 80% methanol and hydro-distillation to obtain the corresponding extracts. Anemonin was isolated from the hydro-distilled extract by preparative TLC. For the in vitro assay, 0.1, 0.4, 2 and 4 mg/ml of the test substances were incubated with parasites and cessation or drop in motility of the parasites was monitored for a total duration of 1 h. In the in vivo assay, the test substances were administered intraperitoneally daily for 7 days to mice infected with Trypanosoma congolense. Diminazene aceturate and 1% dimethylsulfoxide (DMSO) were used as positive and negative controls, respectively. RESULTS: Both extracts showed antitrypanosomal activity although the hydro-distilled extract demonstrated superior activity compared to the hydroalcoholic extract. At a concentration of 4 mg/ml, the hydro-distilled extract drastically reduced motility of trypanosomes within 20 min. Similarly, anemonin at the same concentration completely immobilized trypanosomes within 5 min of incubation, while diminazene aceturate (28.00 mg/kg/day) immobilized the parasites within 10 min. In the in vivo antitrypanosomal assay, anemonin eliminates parasites at all the tested doses (8.75, 17.00 and 35.00 mg/kg/day) and prevented relapse, while in diminazene aceturate-treated mice the parasites reappeared on days 12 to 14. CONCLUSIONS: The current study demonstrated that the fresh leaves of R. multifidus possess genuine antitrypanosomal activity supporting the use of the plant for the treatment of animal trypanosomiasis in traditional medicine. Furthermore, anemonin appears to be responsible for the activity suggesting its potential as a scaffold for the development of safe and cost effective antitrypanosomal agent.

Two undescribed benzopyran derivatives from Ranunculus sceleratus L. with their in vitro anti-inflammatory activity in lipopolysaccharide-activated RAW 264.7 cells.

Previous studies on Ranunculus sceleratus L. have shown the existence of coumarins and their anti-inflammatory effect. Phytochemical work was conducted to investigate the bioactive compounds, leading to the isolation of two undescribed benzopyran derivatives, namely ranunsceleroside A (1) and B (3), together with two known coumarins (2, 4) from the whole plant of R. sceleratus L. All compounds were structurally identified by extensive spectroscopic analysis and then investigated for their inhibitory effect on nitric oxide (NO), tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß) and interleukin-6 (IL-6) production induced by lipopolysaccharide (LPS) in RAW 264.7 murine macrophages, repectively. As a result, compound 1-4 presented inhibitory effects on the production of NO, TNF-α, IL-1ß, and IL-6 in a concentration-dependent manner, which provides a potential chemical basis for the traditional use of R. sceleratus L. as an anti-inflammatory plant.

Ultrahigh-porosity Ranunculus-like MgO adsorbent coupled with predictive deep belief networks: A transformative method for phosphorus treatment.

Phosphorus is a nonrenewable material with a finite supply on Earth; however, due to the rapid growth of the manufacturing industry, phosphorus contamination has become a global concern. Therefore, this study highlights the remarkable potential of ranunculus-like MgO (MO4-MO6) as superior adsorbents for phosphate removal and recovery. Furthermore, MO6 stands out with an impressive adsorption capacity of 596.88 mg/g and a high efficacy across a wide pH range (2-10) under varying coexisting ion concentrations. MO6 outperforms the top current adsorbents for phosphate removal. The process follows Pseudo-second-order and Langmuir models, indicating chemical interactions between the phosphate species and homogeneous MO6 monolayer. MO6 maintains 80 % removal and 96 % recovery after five cycles and adheres to the WHO and EUWFD regulations for residual elements in water. FT-IR and XPS analyses further reveal the underlying mechanisms, including ion exchange, electrostatic, and acid-base interactions. Ten machine learning (ML) models were applied to simultaneously predict multi-criteria (sorption capacity, removal efficiency, final pH, and Mg leakage) affected by 15 diverse environmental conditions. Traditional ML models and deep neural networks have poor accuracy, particularly for removal efficiency. However, a breakthrough was achieved by the developed deep belief network (DBN) with unparalleled performance (MAE = 1.3289, RMSE = 5.2552, R2 = 0.9926) across all output features, surpassing all current studies using thousands of data points for only one output factor. These captivating MO6 and DBN models also have immense potential for effectively applying in the real water test with error < 5 %, opening immense horizons for transformative methods, particularly in phosphate removal and recovery.

Appraisals on the chemical characterization and biological potentials of Ranunculus constantinopolitanus extracts using chromatographic, computational, and molecular network approaches.

In this context, phytochemicals were extracted from Ranunculus constantinopolitanus using ethyl acetate (EA), ethanol, ethanol/water (70%), and water solvent. The analysis encompassed quantification of total phenolic and flavonoid content using spectrophotometric assays, chemical profiling via high performance liquid chromatography-mass spectrometry/mass spectrometry (HPLC-MS/MS) for the extracts, and assessment of antioxidant activity via 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS), Cupric reducing antioxidant capacity (CUPRAC), ferric reducing antioxidant power (FRAP), metal chelating (MCA), and phosphomolybdenum (PBD) assays. Moreover, antimicrobial activity was assessed against four different bacterial strains, as well as various yeasts. Enzyme inhibitory activities were evaluated against five types of enzymes. Additionally, the extracts were examined for their anticancer and protective effects on several cancer cell lines and the human normal cell line. All of the extracts exhibited significant levels of ferulic acid, kaempferol, and caffeic acid. All tested extracts demonstrated antimicrobial activity, with Escherichia coli and Pseudomonas aeruginosa being most sensitive to EA and ethanol extracts. Molecular docking studies revealed that kaempferol-3-O-glucoside strong interactions with AChE, BChE and tyrosinase. In addition, network pharmacology showed an association between gastric cancer and kaempferol-3-O-glucoside. Based on the results, R. constantinopolitanus can be a potential reservoir of bioactive compounds for future bioproduct innovation and pharmaceutical industries.

Experimental investigation of the responses of meadow buttercup (Ranunculus acris L.) to sodic salinity and its implications for habitat monitoring.

Ranunculus acris L. is a native species widely distributed throughout Europe and is invasive in nonnative areas, causing substantial economic losses in pasture productivity. The present study examined the effects of sodic salinity on the growth and functioning of this species. Salinity stresses the germination process and seedling growth, indicating that the studied species experience serious limitations at 60-90 mmol dm-3 NaCl and cannot establish in habitats where salinity is equal to or greater than 150 mmol dm-3 NaCl. R. acris is tuned to subsaline habitats characteristic of temperate meadows, as its growth and functioning were the best when the plants were treated with 30 mmol dm-3 NaCl. Increasing salinity (60 and 90 mmol dm-3 NaCl) hampered growth, leaf morphology and photosynthesis but not mineral nutrition, as Na accumulation seemed to be the most outlined effect of NaCl application. Changes in leaf morphological characteristics coordinated well with Na content in those organs, which indicates that leaf appearance can be easily catchable sign of progressing salinity. Ultimately, progressing salinity reduces the competitiveness of the studied species, shifting its strategy to ruderal behavior, but under subsaline conditions, the strategy of this species seems to be most balanced.

Emergent (branched bur-reed-Sparganium erectum L.) and submergent (river water-crowfoot-Ranunculus fluitans Wimm., 1841) aquatic plants as metal biosorbents under varying water pH conditions in laboratory conditions.

A laboratory experiment was performed to find out the potential bioaccumulation of metals (Al, Cu, Cd, Ni, Pb, Fe, Mn) in aquatic plants acting as natural biosorbents in the environment depending on the variation of water pH. Two plants were selected for the study: branched bur-reed-Sparganium erectum L., as a representative of emergent plants; and river water-crowfoot-Ranunculus fluitans Wimm., 1841, as a representative of submergent plants. The bioaccumulation of metals in plants relative to water (BCFW) was determined, and the metal pollution index (MPI) was calculated. The metal content in water and plants before the experiment was arranged in the following series of increasing values: Cd = Ni < Cu < Pb < Mn < Al < Fe (water) and Cd < Cu < Ni < Pb < Al < Fe < Mn (in both plants). The lowest concentration of cadmium was found in water both at the end and beginning of the experiment. In turn, aluminum level increased, and its concentration was the highest in both acidic and alkaline water. This suggests that aluminum is definitely more available to the organisms living in it under such conditions, and in higher concentrations may be a limiting factor for them. Bioaccumulation of metals (BCFW) in neutral and alkaline environments was higher in branched bur-reed, and in acidic conditions in river water-crowfoot. This may provide a basis for using specific plant species to selectively accumulate metals depending on the range of water pH. This is also supported by the MPI values, which indicate that they have a very high and often the highest impact on metal pollution levels. The observed variability of metals bioaccumulation against the condition of hydromacrophytes makes it possible to consider these plants as biosorbents used in constructed biological treatment plants, which, depending on the species placed there, can selectively absorb particular trace elements. The plants used in the experiment are common hydromacrophytes of Europe, so they can be used in the construction of artificial wetlands across the continent. The apparent diversity of metal accumulation in relation to hydromacrophytes indicates that this type of site can be inhabited by specific species, which can significantly improve the purification of natural or anthropogenic water reservoirs, i.e., those that are intended to perform a filtering function.

Complete Chloroplast Genome Determination of Ranunculus sceleratus from Republic of Korea (Ranunculaceae) and Comparative Chloroplast Genomes of the Members of the Ranunculus Genus.

Ranunculus sceleratus (family: Ranunculaceae) is a medicinally and economically important plant; however, gaps in taxonomic and species identification limit its practical applicability. This study aimed to sequence the chloroplast genome of R. sceleratus from Republic of Korea. Chloroplast sequences were compared and analyzed among Ranunculus species. The chloroplast genome was assembled from Illumina HiSeq 2500 sequencing raw data. The genome was 156,329 bp and had a typical quadripartite structure comprising a small single-copy region, a large single-copy region, and two inverted repeats. Fifty-three simple sequence repeats were identified in the four quadrant structural regions. The region between the ndhC and trnV-UAC genes could be useful as a genetic marker to distinguish between R. sceleratus populations from Republic of Korea and China. The Ranunculus species formed a single lineage. To differentiate between Ranunculus species, we identified 16 hotspot regions and confirmed their potential using specific barcodes based on phylogenetic tree and BLAST-based analyses. The ndhE, ndhF, rpl23, atpF, rps4, and rpoA genes had a high posterior probability of codon sites in positive selection, while the amino acid site varied between Ranunculus species and other genera. Comparison of the Ranunculus genomes provides useful information regarding species identification and evolution that could guide future phylogenetic analyses.

Full-length transcriptome analysis provides insights into flavonoid biosynthesis in Ranunculus japonicus.

Ranunculus japonicus Thunb. is a traditional Chinese herb. Plants in the genus Ranunculus are generally rich in flavonoids, which have antibacterial, anti-infective, and other pharmacological effects. However, owing to the lack of reference genomes, little is known about the flavonoid biosynthetic pathway in R. japonicus. In this study, PacBio isoform sequencing (PacBio iso-seq) and DNA nanoball sequencing (DNB-seq) were combined to build a full-length transcriptome database for three different tissues of R. japonicus. A total of 395,402 full-length transcripts were obtained, of which 308,474 were successfully annotated. A Kyoto Encyclopedia of Genes and Genomes analysis identified 29 differentially expressed genes encoding nine key enzymes for flavonoid biosynthesis. Correlation analysis indicated that flavanone 3-hydroxylase and flavonol synthase genes might have key roles in the accumulation of flavonoid substances in the different tissues of R. japonicus. The structures of chalcone synthase and chalcone isomerase enzymes were spatially modeled. Reverse-transcription quantitative PCR was used to verify gene expression levels of key enzymes associated with flavonoid biosynthesis. In addition, 22 MYB transcription factors involved in flavonoid biosynthesis and phenylpropanoid biosynthesis were discovered. The reliable transcriptomic data from this study provide genetic information about R. japonicus as well as insights into the molecular mechanism of flavonoid biosynthesis. The results also provide a basis for developing the medicinal value R. japonicus.

Origin, genetic structure and evolutionary potential of the natural hybrid Ranunculus circinatus × R. fluitans.

Understanding the genetic variability of hybrids provides information on their current and future evolutionary role. In this paper, we focus on the interspecific hybrid Ranunculus circinatus × R. fluitans that forms spontaneously within the group Ranuculus L. sect. Batrachium DC. (Ranunculaceae Juss.). Genome-wide DNA fingerprinting using amplified fragment length polymorphisms (AFLP) was employed to determine the genetic variation among 36 riverine populations of the hybrid and their parental species. The results demonstrate a strong genetic structure of R. circinatus × R. fluitans within Poland (Central Europe), which is attributed to independent hybridization events, sterility of hybrid individuals, vegetative propagation, and isolation through geographical distance within populations. The hybrid R. circinatus × R. fluitans is a sterile triploid, but, as we have shown in this study, it may participate in subsequent hybridization events, resulting in a ploidy change that can lead to spontaneous fertility recovery. The ability to produce unreduced female gametes of the hybrid R. circinatus × R. fluitans and the parental species R. fluitans is an important evolutionary mechanism in Ranunculus sect. Batrachium that could give rise to new taxa.

Evolution of Transcriptomes in Early-Generation Hybrids of the Apomictic Ranunculus auricomus Complex (Ranunculaceae).

Hybridisation in plants may cause a shift from sexual to asexual seed formation (apomixis). Indeed, natural apomictic plants are usually hybrids, but it is still unclear how hybridisation could trigger the shift to apomixis. The genome evolution of older apomictic lineages is influenced by diverse processes such as polyploidy, mutation accumulation, and allelic sequence divergence. To disentangle the effects of hybridisation from these other factors, we analysed the transcriptomes of flowering buds from artificially produced, diploid F2 hybrids of the Ranunculus auricomus complex. The hybrids exhibited unreduced embryo sac formation (apospory) as one important component of apomixis, whereas their parental species were sexual. We revealed 2915 annotated single-copy genes that were mostly under purifying selection according to dN/dS ratios. However, pairwise comparisons revealed, after rigorous filtering, 79 genes under diversifying selection between hybrids and parents, whereby gene annotation assigned ten of them to reproductive processes. Four genes belong to the meiosis-sporogenesis phase (ASY1, APC1, MSP1, and XRI1) and represent, according to literature records, candidate genes for apospory. We conclude that hybridisation could combine novel (or existing) mutations in key developmental genes in certain hybrid lineages, and establish (together with altered gene expression profiles, as observed in other studies) a heritable regulatory mechanism for aposporous development.

Extensive hybridization in Ranunculus section Batrachium (Ranunculaceae) in rivers of two postglacial landscapes of East Europe.

We demonstrate a wide distribution and abundance of hybrids between the river species Ranunculus aquatilis, R. fluitans and R. kauffmannii with the still water species R. circinatus (Batrachium, Ranunculaceae) in rivers of two postglacial landscapes of East Europe, i.e., Lithuania and Central European Russia. The Batrachium species and hybrid diversity is higher in the rivers of Lithuania (4 species and 3 hybrids vs. 2 and 1) and represented mainly by western R. aquatilis, R. fluitans and their hybrids whereas in Central European Russia, the East European species R. kauffmannii and its hybrid are the only dominant forms. Hybrids make up about 3/4 of the studied individuals found in 3/4 of the studied river localities in Lithuania and 1/3 of the individuals found in 1/3 of the localities in Central European Russia. Such extensive hybridization in river Batrachium may have arisen due to the specificity of rivers as open-type ecosystems. It may have been intensified by the transformation of river ecosystems by human activities and the postglacial character of the studied landscapes combined with ongoing climate change. Almost all hybrids of R. aquatilis, R. fluitans and R. kauffmannii originated from unidirectional crossings in which R. circinatus acted as a pollen donor. Such crossings could be driven by higher frequency and abundance of R. circinatus populations as well as by some biological mechanisms. Two hybrids, R. circinatus × R. fluitans and R. circinatus × R. kauffmannii, were formally described as R. × redundans and R. × absconditus. We found a hybrid which most likely originated from additional crossing between R. aquatilis and R. circinatus × R. fluitans.

Untying Gordian knots: unraveling reticulate polyploid plant evolution by genomic data using the large Ranunculus auricomus species complex.

Speciation via hybridization and polyploidization is a major evolutionary force in plant evolution but is still poorly understood for neopolyploid groups. Challenges are attributed to high heterozygosity, low genetic divergence, and missing information on progenitors, ploidy, and reproduction. We study the large Eurasian Ranunculus auricomus species complex and use a comprehensive workflow integrating reduced-representation sequencing (RRS) genomic data to unravel reticulate evolution, genome diversity and composition of polyploids. We rely on 97 312 restriction site-associated DNA sequencing (RAD-Seq) loci, 576 targeted nuclear genes (48 phased), and 71 plastid regions derived from 78 polyploid apomictic taxa and four diploid and one tetraploid putative sexual progenitor species. We applied (phylo)genomic structure, network, and single nucleotide polymorphism (SNP)-origin analyses. Results consistently showed only 3-5 supported and geographically structured polyploid genetic groups, each containing extant sexual and one unknown progenitor species. Combined analyses demonstrated predominantly allopolyploid origins, each involving 2-3 different diploid sexual progenitor species. Young allotetraploids were characterized by subgenome dominance and nonhybrid SNPs, suggesting substantial post-origin but little lineage-specific evolution. The biodiversity of neopolyploid complexes can result from multiple hybrid origins involving different progenitors and substantial post-origin evolution (e.g. homoeologous exchanges, hybrid segregation, gene flow). Reduced-representation sequencing genomic data including multi-approach information is efficient to delimit shallow reticulate relationships.

Chemical Constituent Analysis of Ranunculus sceleratus L. Using Ultra-High-Performance Liquid Chromatography Coupled with Quadrupole-Orbitrap High-Resolution Mass Spectrometry.

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.

Radix Ranunculi ternati: review of its chemical constituents, pharmacology, quality control and clinical applications.

OBJECTIVES: Radix ranunculi ternati (RRT) is widely used as traditional and folk medicine distributed in China, as well as in Kyushu and Shikoku in Japan. Although RRT has a short history of use as medicine, it has a high medicinal value. This study summarizes the research progress on the chemical constituents, pharmacological activity, quality control measures and clinical applications to provide a valuable and exhaustive reference for the development and application of RRT. KEY FINDINGS: Phytochemical studies showed that this plant mainly contains glycosides, organic acids, esters, sterols, flavonoids, alkaloids and volatile oils. The pharmacological activity of RRT includes immune regulation, anti-tumour, anti-tuberculosis, antioxidant, antibacterial, hepatoprotective and anti-inflammatory effects. This plant is especially effective in the treatment of cancer, tuberculosis, thyroid and nasopharynges disorders.

Ranunculus ternatus Thunb extract attenuates renal fibrosis of diabetic nephropathy via inhibiting SMYD2.

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.

Inhibitory activities of Ranunculus japonicus Thunb. ethanol extract against hepatitis B virus.

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.

Muricazine, a new hydrazine derivative from Ranunculus muricatus L. with antioxidant, lipoxygenase and urease inhibitory activities.

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.