Primary Investigation of Phenotypic Plasticity in Fritillaria cirrhosa Based on Metabolome and Transcriptome Analyses.

Phenotypic plasticity refers to the adaptability of an organism to a heterogeneous environment. In this study, the differential gene expression and compositional changes in Fritillaria cirrhosa during phenotypic plasticity were evaluated using transcriptomic and metabolomic analyses. The annotation profiles of 1696 differentially expressed genes from the transcriptome between abnormal and normal phenotypes revealed that the main annotation pathways were related to the biosynthesis of amino acids, ABC transporters, and plant-pathogen interactions. According to the metabolome, the abnormal phenotype had 36 upregulated amino acids, including tryptophan, proline, and valine, which had a 3.77-fold higher relative content than the normal phenotype. However, saccharides and vitamins were found to be deficient in the abnormal phenotypes. The combination profiles demonstrated that phenotypic plasticity may be an effective strategy for overcoming potential stress via the accumulation of amino acids and regulation of the corresponding genes and transcription factors. In conclusion, a pathogen attack on F. cirrhosa may promote the synthesis of numerous amino acids and transport them into the bulbs through ABC transporters, which may further result in phenotypic variation. Our results provide new insights into the potential mechanism of phenotypic changes.

Discovery of steroidal alkaloid glycosides from the bulbs of Fritillaria unibracteata with anti-inflammatory activities using an in vivo zebrafish model.

Nine undescribed steroidal alkaloid glycosides, unibrasolanosides A-F, unibraverazosides A-B, and unibratomatoside A, were isolated from the bulbs of Fritillaria unibracteata P. K. Hsiao & K. C. Hsia (Liliaceae). Their structures were elucidated by HRESIMS and 1D and 2D NMR data analyses as well as chemical methods and single-crystal X-ray diffraction analyses. Further investigation revealed that eight steroidal alkaloid glycosides displayed moderate anti-inflammatory activity in vivo in a CuSO4-induced transgenic zebrafish model.

Steroidal alkaloids and their glycosides from the bulbs of Fritillaria unibracteata.

Seven undescribed steroidal alkaloids, including two jervine-type steroidal alkaloids, fritiunibras A-B (1-2), and five cevanine-type steroidal alkaloid glycosides, fritiunibras C-G (3-7), along with six known cevanine-type steroidal alkaloids and their glycosides (8-13) were isolated from the bulbs of Fritillaria unibracteata Hsiao et K. C. Hsia. Their structures were determined by interpretation of comprehensive spectroscopic and single-crystal X-ray diffraction analysis. The absolute configurations of sugar moieties were determined by HPLC analysis and compared with standards after hydrolysis and derivatization. Furthermore, their inhibitory effects on NO production and cytotoxic activities were evaluated.

Natural drug sources for respiratory diseases from Fritillaria: chemical and biological analyses.

Fritillaria naturally grows in the temperate region of Northern Hemisphere and mainly distributes in Central Asia, Mediterranean region, and North America. The dried bulbs from a dozen species of this genus have been usually used as herbal medicine, named Beimu in China. Beimu had rich sources of phytochemicals and have extensively applied to respiratory diseases including coronavirus disease (COVID-19). Fritillaria species have alkaloids that act as the main active components that contribute multiple biological activities, including anti-tussive, expectorant, and anti-asthmatic effects, especially against certain respiratory diseases. Other compounds (terpenoids, steroidal saponins, and phenylpropanoids) have also been identified in species of Fritillaria. In this review, readers will discover a brief summary of traditional uses and a comprehensive description of the chemical profiles, biological properties, and analytical techniques used for quality control. In general, the detailed summary reveals 293 specialized metabolites that have been isolated and analyzed in Fritillaria species. This review may provide a scientific basis for the chemical ecology and metabolomics in which compound identification of certain species remains a limiting step.

LC-MS/MS coupled with chemometric analysis as an approach for the differentiation of bulbus Fritillaria unibracteata and Fritillaria ussuriensis.

INTRODUCTION: Bulbus Fritillaria unibracteata Hsiao et K.C. Hsia is an important traditional Chinese medicine, widely used for the treatment of coughs, phlegm and asthma for thousands of years. Due to an increasing demand in clinic practices, a variety of substitutes have appeared in the market, resulting in a big challenge in the differentiation of bulbus F. unibracteata and its substitutes. AIM: To differentiate bulbus F. unibracteata and its substitutes (bulbus Fritillaria ussuriensis Maxim.) based on their main isosteroidal alkaloid contents, and to test the potentiality of chemometrics as a tool for discrimination. MATERIALS AND METHODS: The nine isosteroidal alkaloids in 61 batches of Fritillariae bulbus were simultaneously quantitated by liquid chromatography with tandem mass spectrometry (LC-MS/MS) method. Principal component analysis (PCA) and hierarchical cluster analysis (HCA) were applied to classify the two kinds of Fritillariae bulbus. RESULTS: Quantitative analysis showed that there were differences in the content of the nine alkaloids between two kinds of Fritillariae bulbus. According to the content of nine isosteroidal alkaloids, bulbus of F. unibracteata and F. ussuriensis were successfully distinguished by PCA model. Among these isosteroidal alkaloids, verticine and verticinone may be used as potential chemical markers for the identification and differentiation between the two kinds of Fritillaria bulbus. CONCLUSION: The LC-MS/MS method coupled with PCA would be a powerful strategy to differentiate bulbus F. unibracteata and substitute specimens for quality evaluation and control.

Identification of evolutionary relationships and DNA markers in the medicinally important genus Fritillaria based on chloroplast genomics.

The genus Fritillaria has attracted great attention because of its medicinal and ornamental values. At least three reasons, including the accurate discrimination between various Fritillaria species, protection and sustainable development of rare Fritillaria resources as well as understanding of relationship of some perplexing species, have prompted phylogenetic analyses and development of molecular markers for Fritillaria species. Here we determined the complete chloroplast (CP) genomes for F. unibracteata, F. przewalskii, F. delavayi, and F. sinica through Illumina sequencing, followed by de novo assembly. The lengths of the genomes ranged from 151,076 in F. unibracteata to 152,043 in F. przewalskii. Those CP genomes displayed a typical quadripartite structure, all including a pair of inverted repeats (26,078 to 26,355 bp) separated by the large single-copy (81,383 to 81,804 bp) and small single-copy (17,537 to 17,569 bp) regions. Fritillaria przewalskii, F. delavayi, and F. sinica equivalently encoded 133 unique genes consisting of 38 transfer RNA genes, eight ribosomal RNA genes, and 87 protein coding genes, whereas F. unibracteata contained 132 unique genes due to absence of the rps16 gene. Subsequently, comparative analysis of the complete CP genomes revealed that ycf1, trnL, trnF, ndhD, trnN-trnR, trnE-trnT, trnN, psbM-trnD, atpI, and rps19 to be useful molecular markers in taxonomic studies owning to their interspecies variations. Based on the comprehensive CP genome data collected from 53 species in Fritillaria and Lilium genera, a phylogenomic study was carried out with three Cardiocrinum species and five Amana species as outgroups. The results of the phylogenetic analysis showed that Fritillaria was a sister to Lilium, and the interspecies relationships within subgenus Fritillaria were well resolved. Furthermore, phylogenetic analysis based on the CP genome was proved to be a promising method in selecting potential novel medicinal resources to substitute current medicinal species that are on the verge of extinction.

Isosteroid alkaloids with different chemical structures from Fritillariae cirrhosae bulbus alleviate LPS-induced inflammatory response in RAW 264.7 cells by MAPK signaling pathway.

Isosteroid alkaloids, natural products from Fritillariae Cirrhosae Bulbus, are well known for its antitussive, expectorant, anti-asthmatic and anti-inflammatory properties. However, the anti-inflammatory effect and its mechanism have not been fully explored. In this study, the anti-inflammatory activitives and the potential mechanisms of five isosteroid alkaloids from F. Cirrhosae Bulbus were investigated in lipopolysaccharide (LPS)-induced RAW264.7 macrophage cells. The pro-inflammatory mediators and cytokines were measured by Griess reagent, ELISA and qRT-PCR. The expression of MAPKs was investigated by western blotting. Treatment with the five isosteroid alkaloids in appropriate concentrations could reduce the production of nitric oxide (NO), tumor necrosis factor α (TNF-α) and interleukin-6 (IL-6) in supernatant, and suppressed the mRNA expressions of TNF-α and IL-6. Meanwhile, the five isosteroid alkaloids significantly inhibited the phosphorylated activation of mitogen activated protein kinase (MAPK) signaling pathways, including extracellular signal-regulated kinase (ERK1/2), p38 MAPK and c-Jun N-terminal kinase/stress-activated protein kinase (JNK/SAPK). These results demonstrated that isosteroid alkaloids from F. Cirrhosae Bulbus exert anti-inflammatory effects by down-regulating the level of inflammatory mediators via mediation of MAPK phosphorylation in LPS-induced RAW264.7 macrophages, thus could be candidates for the prevention and treatment of inflammatory diseases.

Isosteroid alkaloids from Fritillaria cirrhosa bulbus as inhibitors of cigarette smoke-induced oxidative stress.

Fritillaria cirrhosa bulbus is a Chinese folk herb famous for its antitussive, expectorant, anti-asthma and anti-inflammatory properties, and is widely used to treat respiratory diseases. However, the impacts of F. cirrhosa bulbus on oxidative stress are still unkown. In the present study, we investigated the potential effect and mechanism of six isosteroid alkaloids with different chemical structures from F. cirrhosa bulbus on protection against cigarette smoke-induced oxidative stress in RAW264.7 macrophages. The results showed that six isosteroid alkaloids reduced reactive oxygen species (ROS) production, elevated glutathione (GSH) level and promoted heme oxygenase (HO-1) expression, which is in association with induction of NF-E2-related factor 2 (Nrf2) nuclear translocation and up-regulation of Nrf2 expression. Among these alkaloids, verticinone, verticine, imperialine-3-ß-D-glucoside, delavine and peimisine exhibited more potent effect against CSE-induced oxidative stress than that of imperialine. These findings for the first time demonstrated that F. cirrhosa bulbus may play a protective role in cellular oxidative stress by activating Nrf2-mediated antioxidant pathway. Furthermore, the differences in antioxidant effects of these alkaloids were compared, as well as the corresponding structure-activity relationships were preliminarily elucidated. This suggested that F. cirrhosa bulbus might be a promising therapeutic treatment for the prevent of oxidative stress-related diseases.

[Comparative Study on Effects of Anti-Gastritis,Gastric Mucosal Protection and Gastrointestinal Movement Promotion of Root and Stem of Zanthoxylum nitidum].

Objective: To provide the experimental evidence for expansion of medicinal parts of Zanthoxylum nitidum by comparing the effects of anti-gastritis,gastric mucosal protection and gastrointestinal movement promotion of its root and stem. Methods: The pharmacological effects between root and stem of Zanthoxylum nitidum were compared by observing the anti-gastritis effect on rats with chronic superficial gastritis induced by iodoacetamide, evaluating the gastric mucosal protective effect on rats' gastric ulcer induced by stress, indometacin and pylorus ligation test, and investigating gastrointestinal movement promotion effect on mice gastric evacuation and intestinal propelling. Results: Both root and stem of Zanthoxylum nitidum showed effects of relieving the inflammation symptoms of rats' gastric mucosa induced by iodoacetamide, gastric ulcer respectively induced by stress, and presenting a strong inhibition of free acid and pepsin activity in gastric juice. Furthermore stem parts of Zanthoxylum nitidum in promoting gastrointestinal motility even showed better efficacy than root. Conclusion: Stem of Zanthoxylum nitidum has similar effects of anti-gastritis, gastric mucosal protection and gastrointestinal movement promotion with root of Zanthoxylum nitidum.

Four new alkylamides from the roots of Zanthoxylum nitidum.

Phytochemical investigation on the roots of Zanthoxylum nitidum led to the isolation of four new unsaturated alkylamides, zanthoxylumamides A-D. Their structures were established via the detailed spectroscopic analyses.