Effect of adulteration on quality of Farfarae Flos / 中草药

Objective: For the adulteration phenomenon of Farfarae Flos, the chemical composition of the flower buds and the rachis, rhizome, and the roots were compared, to provide the basis for the quality control of Farfarae Flos. Methods: The content of tussilagone was determined by high performance liquid chromatography (HPLC) according to the Chinese Pharmacopeia. The HPLC based fingerprint was also generated, and the similarity and the relative contents of the common peaks between the flower buds and adulteration parts were calculated. The pearson correlation between the relative content of the major compounds and the flower buds ratio, as well as principal component analysis and clustering analysis were also performed. Results: The content of tussilagone and the peak areas of 13 common peaks in the HPLC fingerprint were significantly higher than those in the rachis, rhizome, and the roots, and positively correlated with the flower buds ratio. The results of the principal component analysis and clustering analysis showed that the flower buds showed distinct separation with those adulteration parts. In addition, the compounds within the caffeoyl quinic acids and flavonoids showed positive correlations with each other, and the correlations were also observed between different kinds of components. Conclusion: The major compounds of Farfarae Flos were mainly present in the flower buds, and the quality of Farfarae Flos will be greatly affected when there are more impurities such as pedicel, taproot and rhizome in the crude drugs. Currently, there is no impurity in the Chinese pharmacopeia for Farfarae Flos, and the limit of the impurities should be added to guarantee the quality of Farfarae Flos.

Effect of Different Drying Methods on the Chemical Composition of Farfarae Flos / 中国药学杂志

OBJECTIVE: To analyze the effect of different drying methods on the major compounds in Farfarae Flos(FF). METHODS: The content of moisture and tussilagone were determined, and the common peaks in the HPLC fingerprint were calculated and subjected to the principal component analysis. RESULTS: The results showed that the moisture content was the highest when the FF was dried in the shade, and the drying method showed little effect on the content of tussilagone. The results of the principal component analysis showed that the FF dried in the shade was different from those of FF being dried under heat. The relative content of major compounds were the highest for the FF dried in the shade. In addition, the caffeoyl quinic acids and flavonoids were greatly affected after heating, however the heat drying showed little effect on the sesquiterpenoids. Among the different drying temperature, 55 ℃ showed smallest effect on the main components in the FF. CONCLUSION: The components in FF can be protected when drying in the shade, which reveals the scientific basis for the traditional experience of drying. However, in order to facilitate the drying process on a large scale, and minimizing the effect of drying on the compounds in the FF, drying temperature of 55 ℃ is recommended.

Study on Quality Standard for Tussilago farfara Formula Granules / 中国药房

OBJECTIVE： To provide reference for the establishment of quality standard of Tussilago farfara formula granules. METHODS： TLC method was used for qualitative identification of tussilagone in T. farfara formula granules. The content of tussilagone in T. farfara formula granule was determined by HPLC. The determination was performed on a Thermo ODS Hypersil C18 column with the mobile phase consisted of methanol-water （85 ∶ 15， V/V） at the flow rate of 1.0 mL/min. The detection wavelength was set at 220 nm， the column temperature was 25 ℃. Sample size was 20 μL.  RESULTS： TLC spots of tussilagone were clear and well-separated， without interference from negative control. The linear range of tussilagone was 1.39-27.75 μg/mL （r＝0.999 9）. The limits of quantification and detection were 0.153 87 and 0.051 42 μg/mL， respectively. RSDs of precision， stability and reproducibility tests were lower than 2％. The recoveries were 97.12％-103.96％ （RSD＝2.60％， n＝6）. CONCLUSIONS： The method is simple， accurate and reproducible， and suitable for quality control of T. farfara formula granules.

Effects of Flower Buds Extract of Tussilago farfara on Focal Cerebral Ischemia in Rats and Inflammatory Response in BV2 Microglia / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the effects of the flower buds extract of Tussilago farfara Linné (Farfarae Flos; FF) on focal cerebral ischemia through regulation of inflammatory responses in activated microglia.</p><p><b>METHODS</b>Brain ischemia was induced in Sprague-Dawley rats by a transient middle cerebral artery occlusion (tMCAO) for 90 min and reperfusion for 24 h. Twenty rats were randomly divided into 4 groups (n=5 per group): normal, tMCAO-induced ischemic control, tMCAO plus FF extract 300 mg/kg-treated, and tMCAO plus MK-801 1 mg/kg-treated as reference drug. FF extract (300 mg/kg, p.o.) or MK-801 (1 mg/kg, i.p.) was administered after reperfusion. Brain infarction was measured by 2,3,5,-triphenyltetrazolium chloride staining. Neuronal damage was observed by haematoxylin eosin, Nissl staining and immunohistochemistry using anti-neuronal nuclei (NeuN), anti-glial fibrillary acidic protein (GFAP), and anti-CD11b/c (OX42) antibodies in ischemic brain. The expressions of inducible nitric oxide synthase (iNOS), tumor necrosis factor (TNF-α), and hypoxia-inducible factor-1a (HIF-1α) were determined by Western blot. BV2 microglial cells were treated with FF extract or its main bioactive compound, tussilagone with or without lipopolysaccharide (LPS). Nitric oxide (NO) production was measured in culture medium by Griess assay. The expressions of iNOS, COX-2 and pro-inflammatory cytokines mRNA were analyzed by reverse transcription-polymerase chain reaction. The expression of iNOS, and COX-2 proteins, the phosphorylation of ERK1/2, JNK, and p38 MAPK and the nuclear expression of NF-κB p65 in BV2 cells were determined by Western blot.</p><p><b>RESULTS</b>FF extract significantly decreased brain infarctions in ischemic rats (P<0.01). The neuronal death and the microglia/astrocytes activation in ischemic brains were inhibited by FF extract. FF extract also suppressed iNOS, TNF-α, and HIF-1α expression in ischemic brains. FF extract (0.2 and 0.5 mg/mL, P<0.01) and tussilagone 20 and 50 μmol/L, P<0.01) significantly decreased LPS-induced NO production in BV2 microglia through downregulation of iNOS mRNA and protein expression. FF extract and tussilagone significantly inhibited LPS-induced expression of TNF-α, IL-1β, and IL-6 mRNA, and also suppressed the phosphorylation of ERK1/2, JNK and p38 MAPK and the nuclear expression of NF-κB in a dose-dependent manner.</p><p><b>CONCLUSIONS</b>FF extract has a neuroprotective effect in ischemic stroke by the decrease of brain infarction, and the inhibition of neuronal death and microglial activation-mediated inflammatory responses.</p>

Tussilagone suppressed the production and gene expression of MUC5AC mucin via regulating nuclear factor-kappa B signaling pathway in airway epithelial cells

In the present study, we investigated whether tussilagone, a natural product derived from Tussilago farfara, significantly affects the production and gene expression of airway MUC5AC mucin. Confluent NCI-H292 cells were pretreated with tussilagone for 30 min and then stimulated with EGF (epidermal growth factor) or PMA (phorbol 12-myristate 13-acetate) for 24 h or the indicated periods. The MUC5AC mucin gene expression was measured by RT-PCR. Production of MUC5AC mucin protein was measured by ELISA. To elucidate the action mechanism of tussilagone, effect of tussilagone on PMA-induced NF-κB signaling pathway was investigated by western blot analysis. Tussilagone significantly inhibited the production of MUC5AC mucin protein and down-regulated the expression of MUC5AC mucin gene, induced by EGF or PMA. Tussilagone inhibited PMA-induced activation (phosphorylation) of inhibitory kappa B kinase (IKK), and thus phosphorylation and degradation of inhibitory kappa Ba (IκBα). Tussilagone inhibited PMA-induced phosphorylation and nuclear translocation of nuclear factor kappa B (NF-κB) p65. This, in turn, led to the down-regulation of MUC5AC protein production in NCI-H292 cells. These results suggest that tussilagone can regulate the production and gene expression of mucin by acting on airway epithelial cells through regulation of NF-κB signaling pathway.

Comprehensive Quality Evaluation of Flos Farfarae from Different Habitats / 中国中医药信息杂志

Objective To compare the quality of Flos Farfarae from different habitats, and provide basis for the utilization and development of Tussilago farfara. Methods The contents of rutin and tussilagone were determinated by HPLC, the 100-bud dry weight and bud color were weighed and observed. And the data of different samples were compared and statistical analysed. Results The content of rutin, tussilagone and 100-bud dry weight in Flos Farfarae from different place has a significant difference, and there was a significant positive correlation between rutin and tussilagone. Principal component and factor analysis showed that the quality of Flos Farfarae from Yushe, Ningwu, Guangling was better than other areas. Conclusion The quality of Flos Farfarae from different areas is difference, and wild T. farfara in Yushe, Ningwu, Guangling could be used as high quality germplasm.