Chrysanthemum morifolium Ramat extract and probiotics combination ameliorates metabolic disorders through regulating gut microbiota and PPARα subcellular localization.

BACKGROUND: Chrysanthemum morifolium Ramat, a traditional Chinese medicine, has the effects on liver clearing, vision improving, and anti-inflammation. C. morifolium and probiotics have been individually studied for their beneficial effects on metabolic diseases. However, the underlying molecular mechanisms were not completely elucidated. This study aims to elucidate the potential molecular mechanisms of C. morifolium and probiotics combination (CP) on alleviating nonalcoholic fatty liver disease (NAFLD) and the dysregulation of glucose metabolism in high-fat diet (HFD)-fed mice. METHODS: The therapeutic effect of CP on metabolism was evaluated by liver histology and serum biochemical analysis, as well as glucose tolerance test. The impact of CP on gut microbiota was analyzed by 16S rRNA sequencing and fecal microbiota transplantation. Hepatic transcriptomic analysis was performed with the key genes and proteins validated by RT-qPCR and western blotting. In addition, whole body Pparα knockout (Pparα-/-) mice were used to confirm the CP-mediated pathway. RESULTS: CP supplementation ameliorated metabolic disorders by reducing body weight and hepatic steatosis, and improving glucose intolerance and insulin resistance in HFD fed mice. CP intervention mitigated the HFD-induced gut microbiota dysbiosis, which contributed at least in part, to the beneficial effect of improving glucose metabolism. In addition, hepatic transcriptomic analysis showed that CP modulated the expression of genes associated with lipid metabolism. CP downregulated the mRNA level of lipid droplet-binding proteins, such as Cidea and Cidec in the liver, leading to more substrates for fatty acid oxidation (FAO). Meanwhile, the expression of CPT1α, the rate-limiting enzyme of FAO, was significantly increased upon CP treatment. Mechanistically, though CP didn't affect the total PPARα level, it promoted the nuclear localization of PPARα, which contributed to the reduced expression of Cidea and Cidec, and increased expression of CPT1α, leading to activated FAO. Moreover, whole body PPARα deficiency abolished the anti-NAFLD effect of CP, suggesting the importance of PPARα in CP-mediated beneficial effect. CONCLUSION: This study revealed the hypoglycemic and hepatoprotective effect of CP by regulating gut microbiota composition and PPARα subcellular localization, highlighting its potential for therapeutic candidate for metabolic disorders.

Chrysanthemum morifolium attenuates metabolic and alcohol-associated liver disease via gut microbiota and PPARα/γ activation.

BACKGROUND: Metabolic and alcohol-associated liver disease (MetALD) shows a high prevalence rate in liver patients, but there is currently no effective treatment for MetALD. As a typical edible traditional Chinese medicinal herb, the anti-inflammatory, antioxidant, and hepatoprotective properties of water extract of Chrysanthemum morifolium Ramat. (WECM) has been demonstrated. However, its therapeutic effect on MetALD and the associated mechanisms remain unclear. PURPOSE: To investigate the underlying mechanisms of WECM against MetALD. METHODS: We constructed a MetALD rat model following a high-fat & high-sucrose plus alcohol diet (HFHSAD). MetALD rats were treated with WECM at 2.1, 4.2, and 8.4 g/kg/d for six weeks. Efficacy was determined, and pathways associated with WECM against MetALD were predicted through serum and hepatic biochemical marker measurement, histopathological section analysis, 16S rDNA sequencing of the gut microbiota and untargeted serum metabolomics analyses. Changes in genes and proteins in the peroxisome proliferator-activated receptor alpha (PPARα) and gamma (PPARγ) signaling pathways were detected by RT‒PCR and Western blotting. RESULTS: WECM treatment significantly attenuated hepatic steatosis, hyperlipidemia and markers of liver injury in MetALD rats. Moreover, WECM improved vascular endothelial function, hypertension, and systematic oxidative stress. Mechanistically, WECM treatment altered the overall structure of the gut microbiota through maintaining Firmicutes/Bacteroidota ratio and reducing harmful bacterial abundances such as Clostridium, Faecalibaculum, and Herminiimonas. Notably, WECM promoted 15-deoxy-△12, 14-prostaglandin J2 (15d-PGJ2) release and further activated the PPARγ to reduce serum TNF-α, IL-1ß, and IL-6 levels. Additionally, WECM upregulated PPARα and downregulated the levels of CD36 and FABP4 to improve lipid metabolism. CONCLUSION: Our findings provide the first evidence that WECM treatment significantly improved hepatic steatosis, oxidative stress and inflammation in MetALD rats by regulating the gut microbiota and activating the 15d-PGJ2/PPARγ and PPARα signaling pathway.

Identification of key phenolic compounds for alleviating gouty inflammation in edible chrysanthemums based on spectrum-effect relationship analyses.

Edible chrysanthemum is a common food resource for tea and functional foods with potential benefits for human health. Studies have indicated that chrysanthemum has the potential effect on inflammatory diseases, while the effects on gouty inflammation remain underexplored. The present study aimed to investigate the anti-gout activity and characterize the active ingredients of chrysanthemums by using metabolite profiles, in vitro experiments, and spectrum-effect analysis. Results showed that 'Boju' (BJ), 'Hangbaiju' (HBJ), and 'Huaiju' (HJ) exhibited regulatory effects on monosodium urate (MSU)-induced inflammation. At the dose of 50 µg/mL, the inhibitory rates of IL-1ß secretion were 24.53 %, 14.36 %, and 38.10 %, respectively. A total of 32 phenolic compounds were identified or preliminarily assigned in UPLC-Q/TOF-MS analysis. And seven phenolics related to anti-gout activity were identified by spectrum-effect relationships. According to ADME (absorption, distribution, metabolism, excretion) evaluation and experiments verification, luteolin, acacetin-7-O-glucoside, and apigenin-7-O-glucoside were critical constituents potentially associated with the reduction of inflammation in gout. Additionally, these phenolics might be suitable as quality control indicators. This study clarified the anti-gout properties of different cultivars of chrysanthemums and active compounds, providing a theoretical basis for its scientific utilization in functional foods.

Residue Analysis and Dietary Risk Assessment of Pymetrozine in Potato (Solanum tuberosum L.) and Chrysanthemum morifolium (Ramat).

Pymetrozine is used on potato (S. tuberosum) and Chrysanthemum morifolium (C. morifolium) to obtain greater yield and quality. However, pesticide use carries the potential for residues to remain and be detected on harvested crops. Therefore, the aim of this study was to estimate pesticide residues in S. tuberosum and C. morifolium products that are commercially available for human consumption and to assess the associated dietary risks. For this study, a total of 340 samples (200 S. tuberosum samples and 140 C. morifolium samples) were collected randomly from supermarkets and farmer's markets. Residues of pymetrozine in S. tuberosum and C. morifolium were detected by using an established and validated QuECHERS-HPLC-MS / MS method, while a dietary risk assessment of pymetrozine in S. tuberosum and C. morifolium was performed using these data. The detection rates of pymetrozine in S. tuberosum and C. morifolium samples were 92.31% and 98.17%, respectively, with residues not more than 0.036 and 0.024 mg/kg, respectively. Based on these results, the dietary risk assessment indicated that the intake of pymetrozine residues in S. tuberosum and C. morifolium does not pose a health risk. This work improved our understanding of the potential exposure risk of pymetrozine in S. tuberosum and C. morifolium.

Six sesquiterpenoids from the stems and leaves of Chrysanthemum morifolium Ramat and their anti-asthma activities.

Six previously undescribed sesquiterpenoids, chrysanthterpenoids H-M (1-6), were isolated from the stem and leaves of Chrysanthemum morifolium Ramat. Structure elucidation of isolated compounds was unambiguously determined based on extensive 1D and 2D NMR spectroscopic analyses. Furthermore, computational prediction of ECD was used to propose the absolute configurations of the compounds. All compounds were evaluated for their anti-asthma effects on RBL-2H3 cells in vitro. The results showed that Compounds 2 and 3 significantly inhibited the release of ß-aminohexosidase and improved RBL-2H3 degranulation by chromogenic substrate and high-content imaging. These results suggest that Compounds 2 and 3 may exhibit anti-asthma activities.

Anxiolytic effects of Chrysanthemum morifolium Ramat Carbonisata-based carbon dots in mCPP-induced anxiety-like behavior in mice: a nature-inspired approach.

Introduction: Anxiety disorders have emerged as a predominant health concern, yet existing pharmacological treatments for anxiety still present various challenges. Chrysanthemum morifolium Ramat Carbonisata (CMRC) has been utilized in China for approximately 400 years as a therapeutic intervention for anxiety disorders. In this study, a novel type of carbon dots derived from the decoction of Chrysanthemum morifolium Ramat Carbonisata (CMRC-CDs) was identified and isolated, and their morphological structure and functional groups were characterized. Furthermore, the effects of CMRC-CDs on m-chlorophenylpiperazine (mCPP)-induced anxiety-like behaviour in mice were examined and quantified. In order to investigate the potential mechanisms of their anxiolytic effects, concentrations of hypothalamic-pituitary-adrenal (HPA) axis hormones, amino acid neurotransmitters, and monoamine neurotransmitters were measured. Methods: In this study, we synthesized CMRC-CDs and evaluated their potential anti-anxiety effects in a controlled experiment involving 48 male ICR mice. The mice were randomly divided into six groups, treated with CMRC-CDs at different doses for 14 days, and subjected to Open-Field (OF) and Elevated Plus Maze (EPM) tests. Post-behavioral evaluations, blood samples and brain tissues were collected for neurotransmitter and Hypothalamic-Pituitary-Adrenal (HPA) axis hormone quantification via ELISA. Additionally, cytotoxicity of CMRC-CDs was assessed using a Cell Counting Kit-8 (CCK-8) assay on RAW 264.7 cells. Results and Discussion: CMRC-CDs were spherical and homogeneously dispersed, with diameters ranging from 1.4 to 4.0 nm and an abundance of chemical groups on their surface. In the open-field (OF) test, mice pre-treated with CMRC-CDs demonstrated an increased proportion of time spent in the central area and a higher frequency of entries into the central area. In the elevated plus maze (EPM) test, mice pre-treated with CMRC-CDs exhibited a greater number of entries into the open arm and an extended duration spent in the open arm. CMRC-CDs were observed to decrease serum concentrations of corticotropin-releasing hormone (CRH), adrenocorticotropic hormone (ACTH), and corticosterone (CORT). Furthermore, CMRC-CDs were found to increase γ-aminobutyric acid (GABA) and 5-hydroxytryptamine (5-HT) levels, while concurrently reducing glutamic acid (Glu) concentrations in brain tissue. CMRC-CDs demonstrated anxiolytic effects, which may be attributed to their modulation of hormones and neurotransmitters. This finding suggests the potential therapeutic value of CMRC-CDs in the clinical treatment of anxiety disorders.

Identification of CmbHLH Transcription Factor Family and Excavation of CmbHLHs Resistant to Necrotrophic Fungus Alternaria in Chrysanthemum.

Chrysanthemum morifolium Ramat. 'Huaihuang' is a traditional Chinese medicinal plant. However, a black spot disease caused by Alternaria sp., a typical necrotrophic fungus, has a serious damaging influence on the field growth, yield, and quality of the plant. 'Huaiju 2#' being bred from 'Huaihuang', shows resistance to Alternaria sp. bHLH transcription factor has been widely studied because of their functions in growth development, signal transduction, and abiotic stress. However, the function of bHLH in biotic stress has rarely been studied. To characterize the resistance genes, the CmbHLH family was surveyed in 'Huaiju 2#'. On the basis of the transcriptome database of 'Huaiju 2#' after Alternaria sp. inoculation, with the aid of the Chrysanthemum genome database, 71 CmbHLH genes were identified and divided into 17 subfamilies. Most (64.8%) of the CmbHLH proteins were rich in negatively charged amino acids. CmbHLH proteins are generally hydrophilic proteins with a high aliphatic amino acid content. Among the 71 CmbHLH proteins, five CmbHLHs were significantly upregulated by Alternaria sp. infection, and the expression of CmbHLH18 was the most significant. Furthermore, heterologous overexpression of CmbHLH18 could improve the resistance of Arabidopsis thaliana to necrotrophic fungus Alternaria brassicicola by enhancing callose deposition, preventing spores from entering leaves, reducing ROS accumulation, increasing the activities of antioxidant enzymes and defense enzymes, and promoting their gene expression levels. These results indicate that the five CmbHLHs, especially CmbHLH18, may be considered candidate genes for resistance to necrotrophic fungus. These findings not only increase our understanding of the role CmbHLHs play in biotic stress but also provide a basis by using CmbHLHs to breed a new variety of Chrysanthemum with high resistance to necrotrophic fungus.

Rapid screening and in vivo target occupancy quantitative evaluation of xanthine oxidase inhibitors based on drug-target binding kinetics research strategy: A case study of Chrysanthemum morifolium Ramat.

Chrysanthemum morifolium Ramat. is a kind of food and drug dual-use traditional Chinese medicine possessing multiple pharmacological and biochemical benefits. In our study, a rapid and high-throughput method based on Surface plasmon resonance (SPR) biosensor technology was developed and verified for screening potential xanthine oxidase (XOD) inhibitors exemplarily in the Chrysanthemum morifolium Ramat. Coupled with ultra-high performance liquid chromatography-electrospray ionization-tandem mass spectrometry (UPLC-ESI-MS), 14 XOD-binders were identified. In the SPR-based biosensor and molecular docking analysis, most compounds exhibited a strong affinity and binding kinetic property (association rate constant, Kon and dissociation rate constant, Koff) for XOD and could be regarded as potential inhibitors. More importantly, to further accurately assess target occupancy of candidate compounds in vivo, a mathematical model was established and verified involving three crucial intrinsic kinetic processes (Pharmacokinetics, Binding kinetic and Target kinetic). Overall, the proposed screening and assessment strategy could be proved an effective theoretical basis for further pharmacodynamic evaluation.

Fusarium oxysporum infection on root elicit aboveground terpene production and salicylic acid accumulation in Chrysanthemum morifolium.

Underground infection of Fusarium oxysporum resulted in great yield losses in chrysanthemum (Chrysanthemum morifolium Ramat.) industry. However, the effect of F. oxysporum root disease on the terpenes production in above- and below-ground parts of plant is completely unexplored. The aim of this study was to investigate the systematic impact of Fusarium infection underground on the terpene production in aboveground parts of chrysanthemum. Terpene production in above- and below-ground parts was profiled in a time series of post-inoculation by GC-MS. Total terpenes were significantly induced from roots and leaves of Fusarium-infected versus healthy plants. These terpenes included monoterpenes, sesquiterpenes and diterpenes, in which sesquiterpenes were primarily induced in roots and leaves, while monoterpenes were produced only in leaves. Through transcriptome analysis, 8 differentially expressed terpene synthase genes (TPSs) were screened out. The relative expression levels of 8 TPS genes at different developmental stage and tissues indicated the spatial delay of the TPS genes in leaves. The induced terpenes from roots and leaves showed consistency with the expression pattern of TPS genes. The biochemical function of Cm-j-TPS1/2/7 were verified by enzymatic assay. Additionally, it's found that the content of salicylic acid (SA) in root and leaf significantly increased by F. oxysporum infection, suggesting a role of the SA signaling pathway in defense. Together, these results reveal the defense response of above- and below-ground parts of plants to root fungal attack and provide a theoretical basis for the effective prediction and control of F. oxysporum infection in chrysanthemum.

Highly divergent isolates of chrysanthemum virus B and chrysanthemum virus R infecting chrysanthemum in Russia.

BACKGROUND: Chrysanthemum is a popular ornamental and medicinal plant that suffers from many viruses and viroids. Among them, chrysanthemum virus B (CVB, genus Carlavirus, family Betaflexiviridae) is widespread in all chrysanthemum-growing regions. Another carlavirus, chrysanthemum virus R (CVR), has been recently discovered in China. Information about chrysanthemum viruses in Russia is very scarce. The objective of this work was to study the prevalence and genetic diversity of CVB and CVR in Russia. METHODS: We surveyed the chrysanthemum (Chrysanthemum morifolium Ramat.) germplasm collection in the Nikita Botanical Gardens, Yalta, Russia. To detect CVB and CVR, we used RT-PCR with virus-specific primers. To reveal the complete genome sequences of CVB and CVR isolates, metatransciptomic analysis of the cultivars Ribonette, Fiji Yellow, and Golden Standard plants, naturally co-infected with CVB and CVR, was performed using Illumina high-throughput sequencing. The recombination detection tool (RDP4) was employed to search for recombination in assembled genomes. RESULTS: A total of 90 plants of 23 local and introduced chrysanthemum cultivars were surveyed. From these, 58 and 43% plants tested positive for CVB and CVR, respectively. RNA-Seq analysis confirmed the presence of CVB and CVR, and revealed tomato aspermy virus in each of the three transcriptomes. Six near complete genomes of CVB and CVR were assembled from the RNA-Seq reads. The CVR isolate X21 from the cultivar Golden Standard was 92% identical to the Chinese isolate BJ. In contrast, genomes of the CVR isolates X6 and X13 (from the cultivars Ribonette and Fiji Yellow, respectively), were only 76% to 77% identical to the X21 and BJ, and shared 95% identity to one another and appear to represent a divergent group of the CVR. Two distantly related CVB isolates, GS1 and GS2, were found in a plant of the cultivar Golden Standard. Their genomes shared from 82% to 87% identity to each other and the CVB genome from the cultivar Fiji Yellow (isolate FY), as well as to CVB isolates from Japan and China. A recombination event of 3,720 nucleotides long was predicted in the replicase gene of the FY genome. It was supported by seven algorithms implemented in RDP4 with statistically significant P-values. The inferred major parent was the Indian isolate Uttar Pradesh (AM765837), and minor parent was unknown. CONCLUSION: We found a wide distribution of CVB and CVR in the chrysanthemum germplasm collection of the Nikita Botanical Gardens, which is the largest in Russia. Six near complete genomes of CVR and CVB isolates from Russia were assembled and characterized for the first time. This is the first report of CVR in Russia and outside of China thus expanding the information on the geographical distribution of the virus. Highly divergent CVB and CVR isolates have been identified that contributes the better understanding the genetic diversity of these viruses.

Rapid characterisation of xanthine oxidase inhibitors from the flowers of Chrysanthemum morifolium Ramat. Using metabolomics approach.

INTRODUCTION: Hyperuricemia is the key risk factor for gout, in which the elevated uric acid is attributed to the oxidation of hypoxanthine and xanthine to uric acid by xanthine oxidase (XO). Adverse effects of the current treatments lead to an urgent need for safer and more effective alternative from natural resources. OBJECTIVE: To compare the metabolite profile of Chrysanthemum morifolium flower fraction with that of its detannified fraction in relation to XO inhibitory activity using a rapid and effective metabolomics approach. METHODS: Proton nuclear magnetic resonance (1 H-NMR)-based metabolomics approach coupled with multivariate data analysis was utilised to characterise the XO inhibitors related to the antioxidant properties, total phenolic, and total flavonoid contents of the C. morifolium dried flowers. RESULTS: The highest XO inhibitory activity, 1,1-diphenyl-2-picryl hydrazyl (DPPH) radical scavenging activity, total phenolic and flavonoid content with strong positive correlation between them were observed in the ethyl acetate (EtOAc) fraction. Detannified EtOAc showed higher XO inhibitory activity than non-detannified EtOAc fraction. A total of 17 metabolites were tentatively identified, of which three namely kaempferol, 4-hydroxybenzoic acid and apigenin, could be suggested to be responsible for the strong XO inhibitory activity. Additive interaction between 4-hydroxybenzoic acid and apigenin (or kaempferol) in XO inhibition was demonstrated in the interaction assay conducted. CONCLUSION: Chrysanthemum morifolium dried flower-part could be further explored as a natural XO inhibitor for its anti-hyperuricemic potential. Metabolomics approach served as an effective classification of plant metabolites responsible for XO inhibitory activity, and demonstrated that multiple active compounds can work additively in giving combined inhibitory effects.

Production of Virus-Free Chrysanthemum (Chrysanthemum morifolium Ramat) by Tissue Culture Techniques.

Almost all plants in their natural environment are commonly infected by viruses. These viral infections can cause devastating diseases and result in severe yield and economic losses, making viral diseases an important limiting factor for agricultural production and sustainable development. However, these losses can be effectively reduced through the productions and applications of virus-free plantlets. In vitro culture techniques are the most successful approaches for efficient eradication of various viruses from almost all the most economically important crops. Techniques for producing virus-free plantlets include meristem tip culture, somatic embryogenesis, chemotherapy, thermotherapy, electrotherapy, shoot tip cryotherapy, and micrografting. Among them, meristem tip culture is currently the most widely used. Here, we describe a detailed protocol for producing virus-free plantlets of Chrysanthemum morifolium Ramat using tissue culture techniques.

Aqueous extract of Chrysanthemum morifolium Ramat. inhibits RANKL-induced osteoclast differentiation by suppressing the c-fos/NFATc1 pathway.

OBJECTIVE: The flower of chrysanthemum, used worldwide as a medicinal and edible product, has shown various bioactivities, such as anti-inflammatory, antioxidant, anti-tumorigenic, and hepatoprotective activities, as well as cardiovascular protection. However, the effect of Chrysanthemum morifolium Ramat. on the regulation of osteoclast differentiation has not yet been reported. In this study, we aimed to investigate the inhibitory effect of Chrysanthemum morifolium Ramat. water extract (CME) on RANKL-induced osteoclast differentiation in mouse bone marrow-derived macrophages (BMMs). STUDY DESIGN: Bone marrow-derived macrophages (BMMs) isolated from the C57BL/6 J mice. The viability of BMMs was detected with MTT assays. Inhibitory effects of CME on osteoclast differentiation and bone resorption was measured by TRAP staining and Pit assay. Osteoclast differentiation-associated gene expression were assessed by Real-time quantitative polymerase chain reaction. Intracellular signaling molecules was assessed by western blot. RESULTS: CME significantly inhibited osteoclast differentiation in BMMs without cytotoxicity, besides inhibiting MAPK/c-fos and PLCγ2/CREB activation. The inhibitory effects of CME on differentiation-related signaling molecules resulted in significant repression of NFATc1 expression, which is a key transcription factor in osteoclast differentiation, fusion, and activation. CONCLUSION: Our results confirmed the inhibition of RANKL-induced PLCγ2/CREB/c-fos/NFATc1 activation by CME during osteoclast differentiation. The findings collectively suggested CME as a traditional therapeutic agent for osteoporosis, RA, and periodontitis.

Flavonoids and caffeoylquinic acids in Chrysanthemum morifolium Ramat flowers: A potentially rich source of bioactive compounds.

Varieties of chrysanthemums are among the world's most valuable edible ornamental crops. However, the availability and relationship between the bio-chemicals of chrysanthemums and their morphological variations remain unclear. We developed liquid chromatography mass spectrometry to construct a spectral tag library to identify and quantify chemicals of 7 caffeoylquinic acids, 21 flavones and flavonols, 4 carotenoids, and 13 other compounds in 27 cultivars and representative tea of Chrysanthemum morifolium. A correlation analysis found that more acacetin 7-O-galactoside (23) resulted in lighter colored flowers and less acacetin (43) and kaempferol (44) was associated with yellow flowers. Hot-H2O extraction of C. morifolium tea showed that most flavonoids and caffeoylquinic acids dissolved out at 30 min, with 20.977 and 8.958 mg/g GW indicated that C. morifolium, which is used in food and tea, is rich in flavonoids and carotenoids. The results improve our understanding of flavonoid biosynthesis and the mechanisms responsible for flower color.

Anti-inflammatory Dendranacetylene A, a new polyacetylene glucoside from the flower of Chrysanthemum morifolium Ramat.

A new polyacetylene glucoside, Dendranacetylene A (1), and a known compound 8E-decaene-4,6-diyn-1-O-ß-d-glucopyranosyl-(1"→2")-ß-d-glucopyranoside (2) were isolated from the flowers of Dendranthema morifolium (Ramat.) kitam. The chemical structures of these compounds were elucidated by NMR and HR-ESI-MS analysis, and comparing these results with data reported in literatures. Additionally, the anti-inflammatory effects of compounds 1 and 2 were evaluated on RAW264.7 murine macrophage cells induced by lipopolysaccharide (LPS). The two compounds significantly inhibited the NO production in LPS-induced RAW 264.7 murine macrophage cells and exhibited anti-inflammatory effects.

Plasma Lipidomics Reveals Insights into Anti-Obesity Effect of Chrysanthemum morifolium Ramat Leaves and Its Constituent Luteolin in High-Fat Diet-Induced Dyslipidemic Mice.

The Chrysanthemum morifolium Ramat (CM) is widely used as a traditional medicine and herbal tea by the Asian population for its health benefits related to obesity. However, compared to the flowers of CM, detailed mechanisms underlying the beneficial effects of its leaves on obesity and dyslipidemia have not yet been elucidated. Therefore, to investigate the lipidomic biomarkers responsible for the pharmacological effects of CM leaf extract (CLE) in plasma of mice fed a high-fat diet (HFD), the plasma of mice fed a normal diet (ND), HFD, HFD plus CLE 1.5% diet, and HFD plus luteolin 0.003% diet (LU) for 16 weeks were analyzed using liquid chromatography-tandem mass spectrometry (LC-MS/MS) combined with multivariate analysis. In our analysis, the ND, HFD, CLE, and LU groups were clearly differentiated by partial least-squares discriminant analysis (PLS-DA) score plots. The major metabolites contributing to this differentiation were cholesteryl esters (CEs), lysophosphatidylcholines (LPCs), phosphatidylcholines (PCs), ceramides (CERs), and sphingomyelins (SMs). The levels of plasma CEs, LPCs, PCs, SMs, and CERs were significantly increased in the HFD group compared to those in the ND group, and levels of these lipids recovered to normal after administration of CLE or LU. Furthermore, changes in hepatic mRNA expression levels involved in the Kennedy pathway and sphingolipid biosynthesis were also suppressed by treatment with CLE or LU. In conclusion, this study examined the beneficial effects of CLE and LU on obesity and dyslipidemia, which were demonstrated as reduced synthesis of lipotoxic intermediates. These results may provide valuable insights towards evaluating the therapeutic effects of CLE and LU and understanding obesity-related diseases.

Protective effect of Chrysanthemum morifolium Ramat. ethanol extract on lipopolysaccharide induced acute lung injury in mice.

BACKGROUND: To evaluate the effect of Chrysanthemum morifolium Ramat. ethanol extract (CEE) on lipopolysaccharide induced acute lung injury in mice. METHODS: The ninety C57BL/6 J male mice randomly divided into five groups: control, model and CEE (50, 100, 200 mg/kg) groups for 7 days oral administration. At the last administration, all mice except control were intratracheal instilled with lipopolysaccharide (LPS, 3 mg/kg) for establish the acute lung injury. Then lung histopathologic, lung wet/dry weight, white blood cells, lymphocytes, neutrophils were detected. The pro-inflammation cytokine tumor necrosis factor-α (TNF-α), interleukin (IL)-6, anti-inflammatory cytokine transforming growth factor-ß1 (TGF-ß1), IL-10 and the marker of antioxides ability total-antioxidant capacity (T-AOC), malondialdehyde (MDA) in lung tissue were measured. RESULTS: The result showed that CEE could improve lung histopathological injury, reduce the ratio of wet/dry lung weight and lung index, inhibit the increased number of white blood cells, lymphocytes and neutrophils, and reduce the increased levels of TNF-α and IL-6. While CEE also significantly increased the levels of TGF-ß1 and IL-10. Furthermore, CEE also markedly increased the activity of T-AOC, and decreased the contents of MDA with a dose-dependent manner. CONCLUSIONS: The study exhibited that CEE has a potential protective effect on lipopolysaccharide induced acute lung injury in mice, the action mechanism of CEE may through balance of the pro-inflammatory and anti-inflammatory factors, and the oxygen free radicals inhibition.

The flower head of Chrysanthemum morifolium Ramat. (Juhua): A paradigm of flowers serving as Chinese dietary herbal medicine.

ETHNOPHARMACOLOGICAL RELEVANCE: Dietary herbal medicines are widely used for the prevention and treatment of a variety of diseases due to their pharmacological activities in China. Juhua (the flower head of Chrysanthemum morifolium Ramat.), the most representative flower-derived one, which is mainly used for the treatment of respiratory and cardiovascular diseases, shows significant activities, such as antimicrobial, anti-inflammatory, and anticancer, and, neuroprotective, as well as effects on the cardiovascular system. AIMS OF THIS REVIEW: This review aims to provide an overview of the crucial roles of flowers in Chinese dietary herbal medicine, and the pharmaceutical research progress of Juhua (the paradigm of dietary herbal medicine derived from the flower) including its applications in Traditional Chinese medicine and diet, cultivars, phytochemistry, quality control, pharmacology, and toxicity, along with chrysanthemum breeding and biotechnology. METHOD: The information associated with Chinese dietary herbal medicine, flower-derived medicine, dietary flower, and pharmaceutical research of Juhua, was collected from government reports, classic books of Traditional Chinese medicine, the thesis of doctors of philosophy and maters, and database including Pubmed, Scifinder, Web of Science, Google Scholar, China National Knowledge Internet; and others. RESULT: All flower-originated crude medicines recorded in Chinese pharmacopeia and their applications were summarized for the first time in this paper. The edible history and development of flowers in China, the theory of Chinese dietary herbal medicines, as well as flowers serving as dietary herbal medicines, were discussed. Moreover, applications in Traditional Chinese medicine and diet, cultivars, phytochemistry, quality control, pharmacology, and safety evaluation of Juhua, together with chrysanthemum breeding and biotechnology, were summarized in this paper. CONCLUSION: The theory of dietary herbal medicines, which are an important part of the Traditional Chinese medicine system, has a history of thousands of years. Many herbal flowers, serving as dietary herbal medicines, contribute significantly to the prevention and treatment of a variety of diseases for Chinese people. To better benefit human health, more effective supervision practice for dietary herbal medicines is needed. Although various investigations on Juhua have been done, there is a lack of analytical methods for discrimination of cultivar flowers and identification of authenticity. Research on the major compounds with bioactivities, especially those related to its clinical application or healthcare function, as well as their possible mechanize, need be strengthened. More safety evaluation of Juhua should be carried out. The research limitations Juhua is facing exist in all dietary herbal medicine.

Study of the mechanism of change in flavonoid composition in the processing of Chrysanthemum morifolium (Ramat.) Tzvel. 'Boju'.

A form of ß-glucosidase was isolated and purified from fresh Chrysanthemum morifolium (Ramat.) Tzvel. 'Boju' (Boju) and its enzymatic properties explored in this study. The purified enzyme and Boju flavonoids were reacted in a water bath to ascertain the composition of the reactants. Flavonoid glycoside and aglycon concentrations in Boju varied significantly depending on processing method. The concentration of flavonoid glycosides in Boju decreased and flavonoid aglycons increased due to heat-activation of ß-glucosidase which hydrolyzed the flavonoid glycosides in Boju to aglycons.

Chrysanthemum Leaf Ethanol Extract Prevents Obesity and Metabolic Disease in Diet-Induced Obese Mice via Lipid Mobilization in White Adipose Tissue.

This study aimed to elucidate the molecular mechanism of Chrysanthemum morifolium Ramat. against obesity and diabetes, by comparing the transcriptional changes in epididymal white adipose tissue (eWAT) with those of the bioactive compound in C. morifolium, luteolin (LU). Male C57BL/6J mice were fed a normal diet, high-fat diet (HFD), and HFD supplemented with 1.5% w/w chrysanthemum leaf ethanol extract (CLE) for 16 weeks. Supplementation with CLE and LU significantly decreased the body weight gain and eWAT weight by stimulating mRNA expressions for thermogenesis and energy expenditure in eWAT via lipid mobilization, which may be linked to the attenuation of dyslipidemia. Furthermore, CLE and LU increased uncoupling protein-1 protein expression in brown adipose tissue, leading to energy expenditure. Of note, CLE and LU supplements enhanced the balance between lipid storage and mobilization in white adipose tissue (WAT), in turn, inhibiting adipocyte inflammation and lipotoxicity of peripheral tissues. Moreover, CLE and LU attenuated hepatic steatosis by suppressing hepatic lipogenesis, thereby ameliorating insulin resistance and dyslipidemia. Our data suggest that CLE helps inhibit obesity and its comorbidities via the complex interplay between liver and WAT in diet-induced obese mice.