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BACKGROUND: Glehnia littoralis is a medicinal and edible plant species having commercial value and has several hundred years of cultivation history. Polyploid breeding is one of the most important and fastest ways to generate novel varieties. To obtain tetraploids of G. littoralis in vitro, colchicine treatment was given to the seeds and then were screened based on morphology, flow cytometry, and root tip pressing assays. Furthermore, transcriptome analysis was performed to identity the differentially expressed genes associated with phenotypic changes in tetraploid G. littoralis. RESULTS: The results showed that 0.05% (w/v) colchicine treatment for 48 h was effective in inducing tetraploids in G. littoralis. The tetraploid G. littoralis (2n = 4x = 44) was superior in leaf area, leaf thickness, petiole diameter, SPAD value (Chl SPAD), stomatal size, epidermal tissues thickness, palisade tissues thickness, and spongy tissues thickness to the diploid ones, while the stomatal density of tetraploids was significantly lower. Transcriptome sequencing revealed, a total of 1336 differentially expressed genes (DEGs) between tetraploids and diploids. Chromosome doubling may lead to DNA content change and gene dosage effect, which directly affects changes in quantitative traits, with changes such as increased chlorophyll content, larger stomata and thicker tissue of leaves. Several up-regulated DEGs were found related to growth and development in tetraploid G. littoralis such as CKI, PPDK, hisD and MDP1. KEGG pathway enrichment analyses showed that most of DEGs were enriched in metabolic pathways. CONCLUSIONS: This is the first report of the successful induction of tetraploids in G. littoralis. The information presented in this study facilitate breeding programs and molecular breeding of G. littoralis varieties.

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ETHNOPHARMACOLOGICAL RELEVANCE: Glehnia littoralis Fr. Schmidtex Miq. is a well-known perennial herb that is used in traditional medicine in China, Japan and Korea. G. littoralis has the effects of treating the lungs with heat, nourishing yin and blood, and acting as an expectorant. Traditional Chinese medicine (TCM) prescriptions containing G. littoralis have various clinical applications, such as clearing heat, relieving coughs, treating hepatic fibrosis, resolving phlegm, and treating esophagitis. AIM OF THE REVIEW: This paper aims to provide a comprehensive and productive review of G. littoralis, mainly including traditional application, ethnopharmacology, chemical composition, pharmacological activities, and quality control. MATERIALS AND METHODS: Literature search was conducted through the Web of Science, ScienceDirect, Springer Link, PubMed, Baidu Scholar, CNKI, and WanFang DATA by using the keywords "Glehnia littoralis", "Radix Glehniae", "Bei Shashen", "Clinical application", "Chemical composition", "Quality control" and "pharmacological action". In addition, information was collected from relevant ancient books, reviews, and documents (1980-2022). RESULTS: G. littoralis is a traditional Chinese herbal medicine with great clinical value and rich resources. More than 186 components, including coumarins, lignans, polyacetylenes, organic acids, flavonoids, and terpenoids, have been isolated and identified from G. littoralis. The pharmacological activities of more than half of these chemicals are yet unknown. Polyacetylenes and coumarins are the most important bioactive compounds responsible for pharmacological activities, such as antiproliferative, anti-oxidation, anti-inflammatory, antibacterial, antitussive, immune regulation and analgesic. In this study, the progress in chemical analysis of G. littoralis, including thin-layer chromatography (TLC), high-performance liquid chromatography (HPLC), gas chromatography-mass spectrometry (MS), and HPLC-MS, were summarized. CONCLUSION: In this paper reviewed the previous literature regarding ethnopharmacological, phytochemical, pharmacological, and quality evaluation of the processing of G. littoralis was reviewed, providing potential reference information for future investigation and clinical applications. However, research on the relationship between chemical constituents and traditional uses of G. littoralis is lacking, and the comprehensive pharmacological effects and mechanisms of G. littoralis require further detailed exploration. In addition, an efficient method for chemical profiling is still unavailable to obtain potent bioactive markers for quality control. Perfect quality standards, which are also the basis for further drug development of G. littoralis, are urgently needed to ensure its quality and clinical application.

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BACKGROUND: Glehnia littoralis is an economic herb with both medicinal and edible uses. It also has important ecological value and special phylogenetic status as it is a monotypic genus species distributing around beach. Little information on its reproductive biology has been reported so far, which has hindered conservation and application of this species. In this study, we observed morphological changes from buds emergence to seeds formation and internal changes during sporogenesis, gametophyte development and embryo and endosperm development of G. littoralis using paraffin-embedded-sectioning and stereo microscope. RESULTS: The results showed that the stages of internal development events of G. littoralis corresponded to obvious external morphological changes, most of developmental features were consistent with other Apiaceae species. The development of male and female gametophytes was not synchronized in the same flower, however, exhibited temporal overlap. From mid-late April to mid-May, the anther primordial and ovule primordial developed into the trinucleate pollen grain and eight-nuclear embryo sac, respectively. From late-May to mid-July, the zygote developed into mature embryo. In addition, some defects in gynoecium or ovule development and abnormal embryo and endosperm development were found. We induced that the possible causes of abortion in G. littoralis were as follows: nutrient limitation, poor pollination and fertilization, and bad weather. CONCLUSIONS: This study revealed the whole process and morphological characteristics of the development of reproductive organ in G. littoralis, which not only provided important data for the study of systematic and conservation biology, but also provided a theoretical basis for cross breeding.

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The dried root of Glehnia littoralis is a traditional Chinese herbal medicine mainly used to treat lung diseases and plays an important role in fighting coronavirus disease 2019 pneumonia in China. This study focused on the key enzyme gene GlPS1 for furanocoumarin synthesis in G. littoralis. In the 35S:GlPS1 transgenic Arabidopsis study, the Arabidopsis thaliana-overexpressing GlPS1 gene was more salt-tolerant than Arabidopsis in the blank group. Metabolomics analysis showed 30 differential metabolites in Arabidopsis, which overexpressed the GlPS1 gene. Twelve coumarin compounds were significantly upregulated, and six of these coumarin compounds were not detected in the blank group. Among these differential coumarin metabolites, isopimpinellin and aesculetin have been annotated by the Kyoto Encyclopedia of Genes and Genomes and isopimpinellin was not detected in the blank group. Through structural comparison, imperatorin was formed by dehydration and condensation of zanthotoxol and a molecule of isoprenol, and the difference between them was only one isoprene. Results showed that the GlPS1 gene positively regulated the synthesis of coumarin metabolites in A. thaliana and at the same time improved the salt tolerance of A. thaliana. Supplementary Information: The online version contains supplementary material available at 10.1007/s11240-022-02427-w.

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OBJECTIVE To explore the extraction process of volatile oil from the stems， leaves and roots of Glehnia littoralis， analyze the chemical components of the volatile oil from the stems， leaves and roots of G. littoralis， and preliminarily evaluate its in vitro antifungal activity. METHODS Based on the steam distillation method， single factor test and orthogonal experiment were conducted to optimize the extraction method of volatile oil from the stems， leaves and roots of G. littoralis. The chemical components of the volatile oil from the stems， leaves and roots of G. littoralis were identified by using gas chromatography-mass spectrometry （GC-MS) technology and their relative contents were calculated. The antifungal activity of volatile oils from the stems， leaves and roots of G. littoralis against Fusarium solani， Fusarium incarnatum， Fusarium oxysporum， Aspergillus parasiticus and Aspergillus flavus was determined by paper diffusion method. RESULTS The optimal extraction process of G. littoralis was solid-liquid ratio of 1∶15， distillation time of 5 hours， and KCl concentration of 15%. Eleven components were identified from the volatile oil of the stems and leaves of G. littoralis， and a total of eight components were identified from the volatile oil of the roots. Ginsenethinol was a common component in the volatile oil from the stems， leaves and roots of G. littoralis， its contents in the stems and leaves， roots were 38.21% and 74.02%， respectively. The volatile oil from the stems， leaves and roots of G. littoralis had a certain E-mail：zwhjzs@126.com inhibitory effect on F. solani， F. incarnatum， F. oxysporum， A. parasiticus and A. flavus， especially volatile oil from the stems and leaves. CONCLUSIONS There is a significant difference in chemical components of the volatile oil between the roots， stems and leaves of G. littoralis， both of which have certain in vitro antifungal activity.

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OBJECTIVE To establish a quality standard for rice-fired Glehnia littoralis . METHODS Appearance observation ， powder microscopic identification and thin-layer chromatography （TLC）identification were performed for the samples of rice-fired G. littoralis decoction piece. According to the relevant methods stated in 2020 edition of Chinese Pharmacopoeia （part Ⅳ），the contents of moisture ，total ash ，acid-insoluble ash ，water-soluble extract and acid-soluble extract were determined. The contents of psoralen，zanthoxylin，bergapten，imperatorin and isoimperatorin were determined by high performance liquid chromatography （HPLC）. RESULTS The rice-fired G. littoralis decoction pieces were round-like small segments ，slightly rough ，yellow（peeled） or dark yellowish brown （with peel ），special gas and slightly sweet taste. The powder was yellowish white. Under the microscope ， secretions and secretory cells ，ducts，gelatinized starch granules ，ray cells ，parenchyma cells ，etc. could be seen. TLC showed the spots developed clearly. In the chromatogram of the test sample ，there was the same blue fluorescent spot at the corresponding position of the chromatogram of isoimperatorin control sample. The moisture ，total ash ，acid-insoluble ash ，water-soluble extract and ethanol-soluble extract from 9 batches of samples were 5.82％-6.27％，3.19％-3.59％，0.21％-0.27％，24.91％-30.30％ and 20.66％ -25.83％ ，respectively. The linear range of psoralen ，zanthotoxin，bergapten，imperatorin and isoimperatorin were 0.240-2.400，0.320-3.200，0.224-2.240，0.292-2.920，0.208-2.080 µg/mL（all r＞0.999 0）. Limits of quantitation were 0.032 0， 0.030 0，0325 0，0.032 0，0.045 0 µg，respectively. Limits of detection were 0.100 8，0.089 6，0.071 5，0.090 0，0.132 0 µg， respectively. RSDs of prescision ，stability（24 h）and reprodu- cibility tests were less than 3％. Average recoveries were 100.56％ （RSD＝1.36％ ，n＝6），100.73％（RSD＝2.25％ ，n＝6）， 100.36％（RSD＝0.98％，n＝6），98.24％（RSD＝0.40％，n＝6） E-mail：853063968@qq.com and 99.40％（RSD＝0.35％，n＝6），respectively. The contents of above five components were 5.85-13.31，8.63-33.38，6.23- E-mail：shixiaofeng2005@sina.com 15.25，6.12-12.98，5.52-10.77 µg/g，respectively. The total contents were 34.20-83.47 µg/g. CONCLUSIONS It is preliminarily proposed that the moisture ，total ash and acid-insoluble ash should not exceed 7.30％，4.10％，0.30％. The water-soluble extract and ethanol-soluble extract are no less than 21.00％ and 18.00％，respectively. The total content of coumarin should not be less than 52.03 µg/g（with peel ）and 26.34 μg/g（peeled）. Established quality standard can be used for the quality control of rice-fired G. littoralis .

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This study investigated the structural elucidation, anti-radical and immunomodulatory activities of polysaccharides from the roots of Glehnia littoralis. A crude polysaccharide was extracted from the roots of G. littoralis through the ultrasonic-assisted extraction and further purified by DEAE-52 cellulose and Sephadex G-100 gel column, a major polysaccharide fraction named GLP80-1 was obtained. The chemical properties and structure of GLP and GLP80-1 were characterized by acid hydrolysis, methylation analysis, along with high performance gel permeation chromatography, Fourier transform infrared, nuclear magnetic resonance, scanning electron microscope, thermogravimetric analysis and X-ray diffraction. The molecular weight distributions of GLP were determined as 1.89 × 106 and 1.26 × 104 Da. The monosaccharide composition of GLP was glucose, glucuronic acid, galactose and arabinose with molar ratios of 0.91:0.04:0.03:0.02, respectively. The average molecular weight of GLP80-1 was determined as 1.63 × 104 Da. The structure of GLP80-1 was deduced to be a homogenous glucan, comprised a main chain of (1→4)-linked-α-D-Glcp with a single α-D-Glcp branch substituted at C-6. The results of biological activities in vitro showed that GLP and GLP80-1 exhibited free radical scavenging effects, and displayed promotion for the proliferation of mouse spleen lymphocytes and RAW264.7 cells. The data indicated that GLP and GLP80-1 had the potential to be explored as novel natural antioxidant and immunomodulator for application in functional food.

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Glehnia littoralis is a medicinal halophyte that inhabits sandy beaches and has high ecological and commercial value. However, the molecular mechanism of salt adaptation in G. littoralis remains largely unknown. Here, we cloned and identified a non-specific phospholipase C gene (GlNPC3) from G. littoralis, which conferred lipid-mediated signaling during the salt stress response. The expression of GlNPC3 was induced continuously by salt treatment. Overexpression of GlNPC3 in Arabidopsis thaliana increased salt tolerance compared to wild-type (WT) plants. GlNPC3-overexpressing plants had longer roots and higher fresh and dry masses under the salt treatment. The GlNPC3 expression pattern revealed that the gene was expressed in most G. littoralis tissues, particularly in roots. The subcellular localization of GlNPC3 was mainly at the plasma membrane, and partially at the tonoplast. GlNPC3 hydrolyzed common membrane phospholipids, such as phosphotidylserine (PS), phosphoethanolamine (PE), and phosphocholine (PC). In vitro enzymatic assay showed salt-induced total non-specific phospholipase C (NPC) activation in A. thaliana GlNPC3-overexpressing plants. Plant lipid profiling showed a significant change in the membrane-lipid composition of A. thaliana GlNPC3-overexpressing plants compared to WT after the salt treatment. Furthermore, downregulation of GlNPC3 expression by virus-induced gene silencing in G. littoralis reduced the expression levels of some stress-related genes, such as SnRK2, P5SC5, TPC1, and SOS1. Together, these results indicated that GlNPC3 and GlNPC3-mediated membrane lipid change played a positive role in the response of G. littoralis to a saline environment.

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A novel polysaccharide (GLP) with a molecular weight of 1.37 × 105 Da was purified from the roots of G. littoralis. Using monosaccharide composition, methylation analysis, GC-MS, 1D and 2D NMR, the structure of GLP was determined to be a 1 → 4)-α-D-Glcp glycoside linkage, while the terminal group of 1→)-α-D-Glcp was bonded to the main chain via O-6. Then, GLP-NPs were prepared by nano-precipitation method, the particle size of GLP-NPs was 288.4 nm and PDI was 0.340. GLP-NPs-AgNPs were prepared using GLP-NPs as reducing agent. GLP-NPs-AgNPs were characterized by ultraviolet-visible spectrophotometer (UV-Vis), Fourier transform infrared spectroscopy (FT-IR), transmission electron microscope (TEM) and X-ray diffraction (XRD). The yield of GLP-NPs-AgNPs was 38.77%, the particle size was 12.5 nm and the chelation rate of silver nanoparticles with polysaccharides was 67.5%. GLP-NPs-AgNPs had better antioxidant and antibacterial activities than GLP and GLP-NPs. In the present work, a simple and eco-friendly approach for the synthesis of silver nanoparticles (AgNPs) using G. littoralis polysaccharides nanoparticles (GLP-NPs) as reducing agent.

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Phytochemical investigation of the roots of Glehnia littoralis Fr. Schmidt. ex Miq. led to the isolation of 16 known compounds, including three ß-carboline alkaloids (1-3), four phenylpropanoids (4-7), five phenolic acids (8-12), three polyacetylenes (13-15) and one fatty acid (16). The structures of these compounds were elucidated on the basis of spectral analysis and comparison with those reported in literatures. To the best of knowledge, the report of the first ß-carboline alkaloid in the Umbelliferae family. Additionally, compounds 1-5, 9, 10 and 16 have not been reported from any species in Umbelliferae family, compounds 7, 8 and 12 were isolated from the genus Glehnia for the first time and could be of the chemotaxinomic significance and serve as valuable chemotaxonomic makers for G. littoralis. The chemotaxonomic significance of the isolated compounds was summarised.

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Glehnia littoralis is an endangered medicinal plant growing in the coastal ecological environment and plays an important role in coastal ecosystems. The endophytes in the plant have a significant role in promoting plant growth and enhancing plant stress resistance. However, the endophytic bacterial structure associated with halophyte G. littoralis is still not revealed. In this project, the construction and diversity of endophytic bacterial consortium associated with different tissues of G. littoralis were illustrated with high throughput sequencing of the V3-V4 region of the bacterial 16S rRNA. The results resolved that the diversity and richness of endophytic bacteria were significantly higher in root than in leaf and stem. The operational taxonomic units (OTU) analysis demonstrated that the Actinobacteria and Proteobacteria were dominant in all the samples at the phylum level, and Pseudomonas, Bacillus, Rhizobium were the dominant genera. Our results unraveled that the bacterial communities differed among different tissues of G. littoralis. Endophytic bacterial communities in leaf and stem shared more similarity than that in the root. Furthermore, the difference of bacteria community and structure among different tissues were also detected by principal coordinate analysis. Taken altogether, we can conclude that the bacterial communities of different tissues are unique, which could facilitate understanding the diversity of endophytic bacteria in G. littoralis.

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Three Apiaceae species Ledebouriella seseloides, Peucedanum japonicum, and Glehnia littoralis are used as Asian herbal medicines, with the confusingly similar common name "Bang-poong". We characterized the complete chloroplast (cp) genomes and 45S nuclear ribosomal DNA (45S nrDNA) sequences of two accessions for each species. The complete cp genomes of G. littoralis, L. seseloides, and P. japonicum were 147,467, 147,830, and 164,633 bp, respectively. Compared to the other species, the P. japonicum cp genome had a huge inverted repeat expansion and a segmental inversion. The 45S nrDNA cistron sequences of the three species were almost identical in size and structure. Despite the structural variation in the P. japonicum cp genome, phylogenetic analysis revealed that G. littoralis diverged 5-6 million years ago (Mya), while P. japonicum diverged from L. seseloides only 2-3 Mya. Abundant copy number variations including tandem repeats, insertion/deletions, and single nucleotide polymorphisms, were found at the interspecies level. Intraspecies-level polymorphism was also found for L. seseloides and G. littoralis. We developed nine PCR barcode markers to authenticate all three species. This study characterizes the genomic differences between L. seseloides, P. japonicum, and G. littoralis; provides a method of species identification; and sheds light on the evolutionary history of these three species.

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Soil salinity is one of the major environmental stresses affecting plant growth, development, and reproduction. Salt stress also affects the accumulation of some secondary metabolites in plants. Glehnia littoralis is an endangered medicinal halophyte that grows in coastal habitats. Peeled and dried Glehnia littoralis roots, named Radix Glehniae, have been used traditionally as a Chinese herbal medicine. Although Glehnia littoralis has great ecological and commercial value, salt-related mechanisms in Glehnia littoralis remain largely unknown. In this study, we analysed the transcriptome of Glehnia littoralis in response to salt stress by RNA-sequencing to identify potential salt tolerance gene networks. After de novo assembly, we obtained 105,875 unigenes, of which 75,559 were annotated in public databases. We identified 10,335 differentially expressed genes (DEGs; false discovery rate <0.05 and |log2 fold-change| ≥ 1) between NaCl treatment (GL2) and control (GL1), with 5,018 upregulated and 5,317 downregulated DEGs. To further this investigation, we performed Gene Ontology (GO) analysis and the Kyoto Encyclopaedia of Genes and Genomes (KEGG) pathway analysis. DEGs involved in secondary metabolite biosynthetic pathways, plant signal transduction pathways, and transcription factors in response to salt stress were analysed. In addition, we tested the gene expression of 15 unigenes by quantitative real-time PCR (qRT-PCR) to confirm the RNA-sequencing results. Our findings represent a large-scale assessment of the Glehnia littoralis gene resource, and provide useful information for exploring its molecular mechanisms of salt tolerance. Moreover, genes enriched in metabolic pathways could be used to investigate potential biosynthetic pathways of active compounds by Glehnia littoralis.

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BACKGROUND: Glehnia littoralis has been used for traditional Asian medicine, which has diverse therapeutic activities. However, studies regarding neurogenic effects of G. littoralis have not yet been considered. Therefore, in this study, we examined effects of G. littoralis extract on cell proliferation, neuroblast differentiation, and the maturation of newborn neurons in the hippocampus of adult mice. METHODS: A total of 39 male ICR mice (12 weeks old) were randomly assigned to vehicle-treated and 100 and 200 mg/kg G. littoralis extract-treated groups (n = 13 in each group). Vehicle and G. littoralis extract were orally administrated for 28 days. To examine neurogenic effects of G. littoralis extract, we performed immunohistochemistry for 5-bromo-2-deoxyuridine (BrdU, an indicator for cell proliferation) and doublecortin (DCX, an immature neuronal marker) and double immunofluorescence staining for BrdU and neuronal nuclear antigen (NeuN, a mature neuronal marker). In addition, we examined expressional changes of brain-derived neurotrophic factor (BDNF) and its major receptor tropomyosin-related kinase B (TrkB) using Western blotting analysis. RESULTS: Treatment with 200 mg/kg, not 100 mg/kg, significantly increased number of BrdU-immunoreactive (+) and DCX+ cells (48.0 ± 3.1 and 72.0 ± 3.8 cells/section, respectively) in the subgranular zone (SGZ) of the dentate gyrus (DG) and BrdU+/NeuN+ cells (17.0 ± 1.5 cells/section) in the granule cell layer as well as in the SGZ. In addition, protein levels of BDNF and TrkB (about 232% and 244% of the vehicle-treated group, respectively) were significantly increased in the DG of the mice treated with 200 mg/kg of G. littoralis extract. CONCLUSION: G. littoralis extract promots cell proliferation, neuroblast differentiation, and neuronal maturation in the hippocampal DG, and neurogenic effects might be closely related to increases of BDNF and TrkB proteins by G. littoralis extract treatment.

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The coastal herbs Glehnia littoralis have been domesticated as traditional medicines for many centuries. The domestication may have caused changes or declines of cultivated G. littoralis (CGL) relative to wild G. littoralis (WGL). By comparing fruit properties of CGL and WGL, we tested the hypothesis that domesticated G. littoralis have suffered major declines, and human cultivation cannot be sufficient to conserve this species. We collected fruits of CGL and WGL in the Shandong peninsula, China, and compared their buoyancy in seawater, germination potential after seawater immersion, and thousand-grain weights. Float rates of the WGL and CGL fruits were 95.6 (mean) ± 2.6% (standard deviation) and 30.0 ± 7.1%, respectively. The germination potential of CGL was significantly reduced, although the thousand-grain weights of CGL (21.85 ± 0.17 g) were higher than those of the WGL fruits (14.73 ± 0.21 g). These results suggest that the CGL have experienced significant declines relative to the WGL, presumably due to the loss of seawater inundation, selection and dispersal. These declines disfavour the persistence of CGL, and human domestication and cultivation are believed to be insufficient for conserving G. littoralis. Sand coasts where WGL still persists should be designated timely as nature reserves to conserve this species.

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Glehnia littoralis is an important medicinal herb disjunctly distributed at sandy beaches in eastern Asia and western North America. The complete chloroplast genome sequence of G. littoralis is 147,552 bp in length and structurally divided into four distinct regions: two copies of inverted repeat of 18,365 bp separated by a large single copy (LSC) of 93,277 bp and a small single copy (SSC) of 17,545 bp. A total of 129 genes are annotated including 85 protein-coding genes, 36 tRNA gene, and eight rRNA genes. Phylogenetic relationship revealed that G. littoralis is closely related to Angelica dahurica in Apiaceae.

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The purpose of this study was to investigate the anticancer activity of polysaccharide (PGL) from Glehnia littoralis on human lung cancer cell line A549. Based on MTT assay, the results suggested that PGL could significantly reduce A549 cells proliferation in a time- and dose-dependent manner. In addition, PGL displayed an inhibitory activity for the A549 cells migration in Transwell migration assay. The results from both flow cytometry analysis and Hochst 3342 staining of apoptotic cells indicated that PGL could promote apoptosis, and induce cycle arrest of A549 cells. Moreover, immunofluorescence assay elucidated PGL could also down-regulate expression of proliferating cell nuclear antigen (PCNA). Overall, these results showed that PGL exerts a strong anticancer action through inhibiting the A549 cells migration, proliferation and inducing cell apoptosis. It could be a new source of natural anticancer agent against lung cancer with potential value in supplements and medicine.

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<p><b>Background</b>Glehnia littoralis has been used for traditional Asian medicine, which has diverse therapeutic activities. However, studies regarding neurogenic effects of G. littoralis have not yet been considered. Therefore, in this study, we examined effects of G. littoralis extract on cell proliferation, neuroblast differentiation, and the maturation of newborn neurons in the hippocampus of adult mice.</p><p><b>Methods</b>A total of 39 male ICR mice (12 weeks old) were randomly assigned to vehicle-treated and 100 and 200 mg/kg G. littoralis extract-treated groups (n = 13 in each group). Vehicle and G. littoralis extract were orally administrated for 28 days. To examine neurogenic effects of G. littoralis extract, we performed immunohistochemistry for 5-bromo-2-deoxyuridine (BrdU, an indicator for cell proliferation) and doublecortin (DCX, an immature neuronal marker) and double immunofluorescence staining for BrdU and neuronal nuclear antigen (NeuN, a mature neuronal marker). In addition, we examined expressional changes of brain-derived neurotrophic factor (BDNF) and its major receptor tropomyosin-related kinase B (TrkB) using Western blotting analysis.</p><p><b>Results</b>Treatment with 200 mg/kg, not 100 mg/kg, significantly increased number of BrdU-immunoreactive () and DCX cells (48.0 ± 3.1 and 72.0 ± 3.8 cells/section, respectively) in the subgranular zone (SGZ) of the dentate gyrus (DG) and BrdU/NeuN cells (17.0 ± 1.5 cells/section) in the granule cell layer as well as in the SGZ. In addition, protein levels of BDNF and TrkB (about 232% and 244% of the vehicle-treated group, respectively) were significantly increased in the DG of the mice treated with 200 mg/kg of G. littoralis extract.</p><p><b>Conclusion</b>G. littoralis extract promots cell proliferation, neuroblast differentiation, and neuronal maturation in the hippocampal DG, and neurogenic effects might be closely related to increases of BDNF and TrkB proteins by G. littoralis extract treatment.</p>

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PURPOSE: Glehnia littoralis has been reported to have several pharmacological properties but no in vivo reports describing the protective effects of this plant on α-amanitin-induced hepatotoxicity have been published. α-Amanitin is a peptide found in several mushroom species that accounts for the majority of severe mushroom poisonings leading to severe hepatonecrosis. In our previous in vitro study, we found that α-amanitin induced oxidative stress, which may contribute to its severe hepatotoxicity. The aim of this study was to investigate whether Glehnia littoralis acetate extract (GLEA) has protective antioxidant effects on α-amanitin-induced hepatotoxicity in a murine model. METHODS: Swiss mice (n=40 in all groups) were divided into four groups (n=10/group). Three hours after giving α-amanitin (0.6 mg/kg, i.p.) to the mice, they were administered silibinin (50 mg/kg/d, i.p.) or Glehnia littoralis ethyl acetate extract (100 mg/kg/d, oral) therapies once a day for 3 days. After 72 hours of treatment, each subject was killed, cardiac blood was aspirated for hepatic aminotransferase measurement, and liver specimens were harvested to evaluate the extent of hepatonecrosis. The degree of hepatonecrosis was assessed by a pathologist blinded to the treatment group and divided into 4 categories according to the grade of hepatonecrosis. RESULTS: GLEA significantly improved the beneficial functional parameters in α-amanitin-induced hepatotoxicity. In the histopathological evaluation, the toxicity that was generated with α-amanitin was significantly reduced by GLEA, showing a possible hepatoprotective effect. CONCLUSION: In this murine model, Glehnia littoralis was effective in limiting hepatic injury after α-amanitin poisoning. Increases of aminotransferases and degrees of hepatonecrosis were attenuated by this antidotal therapy.

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Objective: To clone the full-length cDNA sequences of psoralen synthase (PS) genes from Glehnia littoralis so as to perform the bioinformatic and expression pattern analysis. Methods: Based on our previous transcriptome sequencing data of G. littoralis, the gene sequences GlPS1 and GlPS2 with high expression level were screened. The 3’cDNA ends of GlPS1 and GlPS2 genes were cloned by the RACE (rapid amplification of cDNA ends) method and the full-length cDNA of genes were assembled by using DNAMAN software. And then encoded GlPS proteins were analyzed by the bioinformatics tools. The issue specific expression of GlPS1 and GlPS2 genes were detected using qPCR. Results: The full-length cDNA of GlPS1 gene was 1 885 bp, which encoding a protein of 495 amino acids with a predicted molecular weight of 55 740.7 and isoelectric point of 8.28; The full-length cDNA of GlPS2 gene was 1 971 bp, which encoding a protein of 502 amino acids with a predicted molecular weight of 56 363.9 and isoelectric point of 6.62. GlPS1 and GlPS2 proteins belong to the cytochrome P450 superfamily, which share one transmembrane zone acting as hydrophilic protein. Phylogenetic analysis showed GlPS1 and GlPS2 were genetically closely related to the PS of Pastinaca sativa, Apium graveolens, Ammi majus. Higher expression of GlPS1 gene was observed in roots than leaves. However, GlPS2 gene was expressed at a relatively higher level in flowers than in roots. Conclusion: The full-length cDNA of GlPS1 and GlPS2 genes were obtained and the expression patterns were explored in G. littoralis for the first time, which provided a foundation for further studies on gene function and genetic regulatory mechanism of GlPS.