Efficacy, chemical composition, and pharmacological effects of herbal drugs derived from Fritillaria cirrhosa D. Don and Fritillaria thunbergii Miq.

Fritillaria cirrhosa D. Don and F. thunbergii Miq. belong to the genus Fritillaria within the Liliaceae family. They are used in traditional Chinese medicines that are often administered in clinical settings as they have notable effects on cough, bronchitis, pneumonia, lung injury, cancer, and other diseases. In this review, we focus on the history, origin, similarities, and differences in efficacy, chemical composition, and pharmacological outcomes of the drugs obtained from F. cirrhosa (FRC) and F. thunbergii (FRT). We list various valuable pharmacological effects of FRC and FRT, including antitussive, expectorant, anti-inflammatory, antioxidant, and anticancer effects. Thus, this review offers a basis for the medical application of and further research into the pharmacological impacts of these two drugs. We believe that new drugs derived from the phytoconstituents of F. cirrhosa and F. thunbergii that have specific therapeutic properties can be developed in the future.

Mechanism of Thunberg Fritillaria in treating endometriosis based on network pharmacology and the effect of Peiminine on the MEK/ERK pathway.

OBJECTIVE: To explore the mechanism of Thunberg Fritillaria in treating endometriosis (EMs) based on network pharmacology and the effect of Peiminine on the MEK/ERK pathway. METHODS: We applied Chinese medicine system pharmacology analysis platform (TCMSP) database and literature search to screen the main chemical components of Fritillaria thunbergii Miq and created a Vanny map from the databases of TCMSP, GENECARDS, Online Mendelian Inheritance in Man (OMIM), and some others. The STRING database was used to construct the protein interaction network of Fritillaria thunbergii Miq and EMs. The overlapping targets and enriched pathways were discovered using the cells of the innate immune annotation database (DAVID) and the Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses. To test the mechanism of Peiminine, the active ingredients of Fritillaria thunbergii, in the therapy of EMs, we designed cell assays and animal research. EMs mouse models were treated with several therapies, including fibrosis inhibitor in Peiminine by utilizing Hematoxylin-eosin staining (HE staining), MASSON staining, Immunohistochemistry, Immunofluorescence, quantitative real-time PCR (qRT-PCR) experiment, and Western blotting test. We evaluated the anti-endometriotic effects of Peiminine using 12Z human endometriotic cells. Cell Counting Kit 8 was used to assess the vitality of 12z cells (CCK8). We evaluated the migration ability of 12z cells by cell scratch test. RESULTS: The effective active ingredients of Fritillaria thunbergii Miq in the treatment of EMs are Pelargonidin, Beta-sitosterol syringaresinol, Peimisine Pelargonidin-3, 5-diglucoside Ziebeimine Zhebeiresinol Verticine Solatubin OSI-2040 Chaksine Peiminine Peiminoside Peiminoside_qt, and 6-Methoxyl-2-acetyl-3-methyl-1, 4-naphthoquinone-8-O-beta-D-glucopyranoside. The critical targets for Fritillaria thunbergii Miq treating EMs are NOS2/PTGS1/AR/PPARG/PTGS2/NCOA2/RXRA/PGR/NR3C1/NCOA1/SLC6A4/OPRM1/BCL2 and ESR1. The results of GO function and KEGG enrichment analysis showed that the role pathway was estrogen-related signaling and thyroid hormone-related signaling. The expression of E-cadherin was decreased in EMs while MEK1/2, P-ERK, N-cadherin and vimentin were all increased in MASSON, immunofluorescence, Real-time PCR and Western blotting. In epithelial 12Z cells, high concentrations of Peiminine can block cell activity and migration, which is directly related to blocking cell fibrosis. CONCLUSION: Overall, this study partially verified the network pharmacological prediction that Peiminine regulates the MAPK pathway in inhibiting 12Z cell proliferation and migration, and finally protects against EMs.

A new alkaloid with cytotoxic activity from Fritillaria thunbergii Miq.

A new alkaloid named zhebeisine (1), together with four known compounds, eduardine (2), zhebeirine (3), isoverticine (4), and verticine (5), was isolated from the bulbs of Fritillaria thunbergii Miq. The structure of the new compound was elucidated on the basis of extensive spectroscopic methods and the in vitro biological activities of it were evaluated as well. Compound 1 features a veratramine skeleton with a rare 6/6/5/6/6/6 fused-ring system, representing the first reported veratramine-type alkaloid with a new oxazinane ring (ring-F) in Fritillaria genus. The cytotoxic activities study revealed that compound 1 inhibited the cell proliferation of HT29 and DLD1 (IC50 values of 25.1 and 48.8 µM, respectively) and also induced apoptosis of the above-mentioned two cancer cell lines.[Formula: see text].

Cloning and Functional Characterization of Full-length cDNA Encoding Squalene Synthase from Fritillaria thunbergii Miq / 中国生物化学与分子生物学报

Squalene is a key metabolic intermediate for sterols and various other triterpenoids. Its biosynthesis is catalyzed by squalene synthase (SQS), which converts two molecules of farnesyl pyrophosphate to squalene. The biosynthetic pathway of Fritillaria thunbergii Miq isosteroid alkaloids is similar to that of triterpenoids. In this study, a full-length cDNA of squalene synthase from Fritillaria thunbergii Mig (FtSQS) was cloned using rapid amplification from cDNA ends (RACE) technology. GenBank accession number was KF551097. 2. Bioinformatics methods were used to characterize the FtSQS in detail, including the detection of conserved regions, sequence homology analysis, secondary and tertiary structure prediction, and phylogenetic tree analysis. The results showed that its open reading frame (ORF) was 1 230 bp and encoded 409 amino acids. Protein-Blast alignment found that amino acid homology with SQS of Indian pine, Truncate alfalfa, Purple shirt, Potato, Bupleurum, Golden iron lock and Arabidopsis reached 73. 84%, 73. 23%, 72. 24%, 70. 66%, 70. 66%, 69. 44%, 68. 14%. Promoter analysis indicated that the 5' upstream region of FtSQS possessed various potential elements associated with physiological and environmental factors. To obtain a soluble recombinant protein, 24 hydrophobic amino acids were deleted from the carboxyl terminus, and the C-terminal truncated mutant FtSQS (FtSQSATM) was expressed in E. coli BL21 (DE3). SDS-PAGE analysis suggested that approximately 66 kD recombinant protein was checked. The in vitro enzymatic reaction proved that FtSQS could catalyze farnesyl pyrophosphate to generate squalene. Expression level of FtSQS mRNA in leaves was the highest, followed by stem and root, but in bulb was much lower than that in other tissues. It suggests that leaves are active organ for biosynthesis of peimine. The identification and function of FtSQS provides an important basis for the study of secondary metabolites of Fritillaria thunbergii Miq.

Metabolic profiling investigation of Fritillaria thunbergii Miq. by gas chromatography-mass spectrometry.

Thunberg fritillary bulb (the dry bulbs of Fritillaria thunbergii Miq.), a traditional Chinese Medicine, is widely applied as an expectorant and antitussive. In this investigation, the primary metabolites of bulbs, flowers, leaves, and stems of F. thunbergii were analyzed by gas chromatography-mass spectrometry. Principal component analysis, partial least squares-discriminate analysis, orthogonal projection to latent structures-discriminate analysis, and heat map analysis showed that there were dissimilar metabolites, and a negative correlation between amino acids and saccharides in different analytes. Furthermore, carbodiimide, tryptophan, glucose-6-phosphate, xylose, 2-piperidinecarboxylic acid, monoamidomalonic acid, phenylalanine, and histidine were found to play an important role in the plant metabolism net of F. thunbergii.

Molecular mechanism of Danggui Beimu Kushen Pills in treating prostatic diseases based on TCM-IP / 中草药

Objective To analyze the material basis and molecular mechanisms of Danggui Beimu Kushen (DBK) Pills in treating prostatic diseases based on the method of integrated pharmacology. Method The platform of Integrative Pharmacology of Traditional Chinese Medicine (TCM-IP, www.tcmip.cn) was utilized to predict the main active ingredients and functional targets of DBK Pills in treating prostatic disease, key targets were screened for enrichment analysis of pathways, and the network of “herb-core component-key target-main pathway” was constructed, and the possible mechanisms of DBK Pills in treating prostatic diseases were explored. Results A total of 532 candidate key targets for the treatment of prostatic diseases by DBK Pills were predicted, and 1 840 terms of gene function and 194 signal pathways were analyzed by gene ontology (GO) and KEGG, respectively. The network analysis of “herb-core component-key target-main pathway” showed that 65 core components were predicted, including 29 ingredients from Angelica sinensis, 11 from Fritillaria thunbergii and 26 from Sophora flavescens. Those predicted components acted on the key targets of prostatic diseases, such as transcription factor binding, negative regulation of apoptosis, et al, through the estrogen, apoptosis, chemokines and other signal pathways, and thus played a role in the regulation of cell cycle, apoptosis and proliferation imbalance, which might be the molecular mechanisms of DBK Pills for the treatment of prostatic disease. Conclusion DBK Pills regulate the development of BPH, prostate cancer and other diseases through multiple pathways with multi-component interacting with multiple targets.

Influence of sulfur-fumigation on pharmacokinetic of peimine and peiminine of Fritillaria thunbergii in rat / 中草药

Objective: To study the influence of sulfur-fumigation on pharmacokinetic of peimine and peiminine of Fritillaria thunbergii in rat plasma by LC-MS/MS. Methods: After random grouping, 18 SD rats were given the solution of fresh-cut and sulfur-fumigated F. thunbergii by ig administration. The blood drug concentration of peimine and peiminine in rat plasma was determined by HPLC-MS/MS, and the pharmacokinetic parameters were calculated with 3P97 software. Results: The pharmacokinetic parameters of peimine and peiminine of sulfur-fumigated F. thunbergii in rat plasma were (66.40 ± 4.65), (146.72 ± 10.88) ng/mL for Cmax, and (181.79 ± 7.85), (457.38 ± 58.81) ng∙h/mL for AUC, respectively. Those of fresh-cut sample in rat plasma were (186.37 ± 18.8), (227.65 ± 7.01) ng/mL for Cmax, and (197.70 ± 18.69), (566.16 ± 41.55) ng∙h/mL for AUC, respectively. The pharmacokinetic parameters of perimine and peiminine of sulfur-fumigated sample in rat plasma were less than those of fresh-cut sample. Conclusion: The results showed that sulfur-fumigation decreased the bioavailability of peimine and peiminine. This study could provide a basis for further clarifying the influence of sulfur-fumigation on efficacy material base of F.thunbergii.

Optimization of Supercritical Fluid Extraction of Total Alkaloids, Peimisine, Peimine and Peiminine from the Bulb of Fritillaria thunbergii Miq, and Evaluation of Antioxidant Activities of the Extracts.

Supercritical fluid extraction (SFE) was used to extract total alkaloids, peimisine, peimine and peiminine from the bulb of Fritillaria thunbergii Miq. The antioxidant capacity of the extracts was evaluated by DPPH radical scavenging activity (DPPH-RSA), ABTS radical scavenging activity (ABTS-RSA) and ferric reducing capacity (FRAP) assay. A central composite design (CCD) with four variables and five levels was employed for optimization of process parameters, and response surface plots were constructed in accordance with a second order polynomial model. Under optimal conditions of 3.0 h, 60.4 °C, 26.5 MPa and 89.3% ethanol, the highest yields were predicted to be 3.8 mg/g for total alkaloids, 0.5 mg/g for peimisine, 1.3 mg/g for peimine and 1.3 mg/g for peiminine, and the antioxidant capacity of extracts displayed EC50, DPPH value of 5.5 mg/mL, EC50, ABTS value of 0.3 mg/mL and FRAP value of 118.2 mg ascorbic acid equivalent (AAE)/100 g.

Element analysis and characteristic identification of non-fumigated and sulfur-fumigated Fritillaria thunbergii Miq. using microwave digestion-inductively coupled plasma atomic emission spectrometry combined with Fourier transform infrared spectrometry.

BACKGROUND: Sulfur-fumigation may induce chemical transformation of traditional Chinese medicines leading to harmful effects following patient ingestion. For quality control, it is urgently needed to develop a reliable and efficient method for sulfur-fumigation identification. MATERIALS AND METHODS: The spectrochemical identification of non-fumigated and sulfur-fumigated Fritillaria thunbergii Miq. was carried out to evaluate inorganic elements and organic components. The concentrations of 12 elements, including Zn, Mn, Cu, Fe, Li, Mg, Sr, Pb, As, Cd, Hg, and S of samples were determined by microwave digestion - inductively coupled plasma atomic emission spectrometry (ICP-AES). Meanwhile, Fourier transform infrared spectrometry (FTIR) was used for the study of chemical group characteristic reactions after sulfur-fumigation. RESULTS: The concentrations of Fe, Mg, Hg, and S elements showed significant differences between non-fumigated and sulfur-fumigated Fritillaria thunbergii Miq. The characteristic stretching vibrations of some groups in FTIR spectra, such as -OH, -S = O and -S-O, provided the identification basis for the discrimination of non-fumigated and sulfur-fumigated Fritillaria thunbergii Miq. CONCLUSION: The application of microwave digestion - ICP-AES was successfully used in combination with FTIR to authenticate and evaluate the quality of medicinal Fritillaria thunbergii Miq. Further applications of this technique should be explored.

Specific PCR identification of Fritillaria thunbergii / 中草药

Objective: To identify Fritillaria thunbergii using special primer by specific PCR. Methods: Gene footprint of F. thunbergii named ZB1 was screened from RAPD amplification. Reclaimed ZB1 gene was inserted into T-vector to be cloned and sequenced. One pair of specific primers P2/P3 were designed according to the ZB1 sequence, and applied in specific PCR reaction using genome DNA of F. thunbergii as template. Results: A specific band around 750 bp was detected in F. thunbergii, while nothing appeared in other varieties. Conclusion: The method is convenient, reproducible, and precise, with broad application prospects.

Clinical Application of Fritillaria thunbergii Miq. Granule in Refractory Acute Leukemia during Perichemotherapy / 中国中医药信息杂志

Objective To translate the findings from fundamental research into clinical application and to evaluate the clinical efficiency and safety of Fritillaria thunbergii Miq. granule as adjunctive means of chemotherapy during peri chemotherapy of refractory acute leukemia. Methods Patients in multiple hospitals were randomly divided into two groups with Fritillaria thunbergii Miq. granule treatment or synchronous control at three days before chemotherapy respectively, according to random approaches for medical treatment. The clinical therapeutic effects were then determined after one course of treatment. Results According to the research project, 138 patients were analyzed statistically, 72 in Fritillaria thunbergii Miq. granule group and 66 in control group. The complete remission rate (CR) of Fritillaria thunbergii Miq. granule group and control group were 36.8% and 25.8% respectively, while the total effects were 77.8% and 53.0%, which was significantly different (P

AFLP Analysis on genetic diversity for germplasm resources of Fritillaria thunbergii cultivated in Zhejiang Province / 中草药

Objective To study the genetic diversity of Fritillaria thunbergii,a traditional Chinese herb in Zhejiang Province in China.Methods The genetic diversity of six representational populations of F.thunbergii including 32 individuals was investigated by amplified fragment length polymorphism(AFLP) maker technique.Results The genetic diversity was revealed as follow: the Nei′s genetic diversity index(He) 0.169 0?0.175 7,Shannon′s information index(I) 0.269 8?0.245 3,percentage of polymorphic loci(PPB) was 76.85% at the species level;Ht 0.169 0?0.030 9,and Hs 0.150 8?0.024 0,I 0.233 3?0.261 9, PPB was 50.38% at population level.The genetic differentiation index(Gst) was 0.107 6,Nm 4.147 0.The result of dendrogram of six populations indicated that Dongyang and Yongkang populations shared the minimum genetic distance(0.015 0),they were classified into a group,and Xiangshan and Jinyun populations shared the maximum genetic distance(0.032 4).Conclusion The genetic diversity of F.thunbergii cultivated in Zhejiang Province is very rich,which could ensure the long-term survival of F.thunbergii.But the genetic diversity of F.thunbergii is relatively higher in population levels while lower at the species levels and the degree of genetic differentiation occured among the populations is not significant.The germplasm resources are relatively stable among these six populations.These populations could be used to breed the fine strains of F.thunbergii as the bases.

Direct identification of Fritillaria cirrhosa, F. ia thunbergii and F. thunbergii var. chekiangensis by FTIR / 中草药

Object To identify Fritillaria cirrhosa D. Don., Fritillaria thunbergii Miq. and Fritillaria thunbergii Miq. var. chekiangensis Hsiao et K. C. Hsia. with FTIR.Methods Their IR spectra were obtained by direct FTIR.Results The infrared spectra of F. cirrhosa, F. thunbergii, F. thunbergii var. chekiangensis were different.Conclusion F. cirrhosa, F. thunbergii, and F. thunbergii var. chekiangensis were identified by FTIR directly, fastly and accurately.