Study on the anti-pulmonary fibrosis effect of linarin in vivo and in vitro and its mechanism / 中国药房

OBJECTIVE To investigate the anti-pulmonary fibrosis effect of linarin in vivo and in vitro， and investigate its mechanism preliminarily. METHODS C57BL/6J mice were randomly divided into normal group （carboxymethylcellulose sodium）， model group （carboxymethylcellulose sodium）， positive control group （pirfenidone， 200 mg/kg）， linarin low-dose and high-dose groups （12.5， 25 mg/kg）， with 8 mice in each group. Except for normal group， pulmonary fibrosis model was induced in other groups. After modeling， they were given relevant medicine intragastrically， once a day， for consecutive 14 d. The general situation of mice was observed， and their lung indexes were measured； the levels of tumor necrosis factor-α （TNF-α） and transforming growth factor-β1（ TGF-β1） in serum and interleukin-6 （IL-6） in lung tissue were determined. Hematoxylin-eosin （HE） staining and Masson staining were used to observe the histopathological morphology of lung. The pulmonary fibrosis was scored according to Ashcroft score standard. The expressions of α-smooth muscle actin （α-SMA） and （type Ⅰ collagen， Collagen Ⅰ）， phosphorylated extracellular signal-regulated kinase （p-ERK1/2） and TGF-β1 in lung tissues were detected. HFL1 cells were stimulated by TGF- β1 to form pulmonary fibrosis model in vitro， which were divided into normal group， model group and linarin low-， medium- and high-concentration groups （3.7， 7.4， 14.8 mg/L）. After being cultured for 48 h， the protein expressions of α-SMA， Collagen Ⅰ and p-ERK1/2 in HFL1 cells were detected. RESULTS In vivo， compared with normal group， the lung index of model group and the levels of TNF- α， TGF- β1 and IL-6 were significantly increased （P＜0.01）. There were a large number of inflammatory infiltration and cellular fibrosis lesions in the alveoli， and a large number of collagen depositions. The scores of HE staining and Masson staining were significantly increased （P＜0.01）. The protein expressions of α-SMA， Collagen Ⅰ， p-ERK1/2 and TGF-β1 in lung tissue were up-regulated significantly （P＜0.01）. Compared with model group， above indexes of mice were improved significantly in linarin high-dose group （P＜0.05 or P＜0.01）， and most of indexes （except for lung index） were improved significantly in linarin low-dose group （P＜0.05 or P＜0.01）. In vitro， compared with blank group， the density of cells in the model group increased， and obvious proliferation and other changes occurred； protein expressions of α-SMA， Collagen Ⅰ and p-ERK1/2 were significantly up-regulated （P＜0.05 or P＜0.01）. Compared with model group， the cell density of each concentration group was decreased and the morphology gradually returned to normal； the expressions of above proteins in linarin high-concentration group and the protein expression of p-ERK1/2 in linarin medium-concentration group were down-regulated significantly（P＜0.05 or P＜0.01）. CONCLUSIONS Linarin may regulate ERK and inflammatory pathways to reduce the inflammatory response， thereby exerting anti-pulmonary fibrosis effect.

Anti-diabetic effects of linarin from Chrysanthemi Indici Flos via AMPK activation / 中草药·英文版

Objective: The purpose of this study is to investigate the anti-diabetic effects of linarin, a flavonoid extracted from Chrysanthemi Indici Flos (CIF), and its potential mechanisms. Methods: The effects of linarin on cell viability and glucose consumption in HepG2 cells were measured. Meanwhile, monosodium glutamate (MSG) mouse model was constructed to monitor the changes of insulin tolerance, glucose tolerance, triglyceride and cholesterol. The protein expression levels of p-AMPK, p-ACC, PEPCK and p-GS were detected by Western blot. Results: Linarin could increase the relative glucose consumption of HepG2 cells, improve insulin tolerance and glucose tolerance, and decrease the levels of triglyceride and cholesterol of MSG mice. Simultaneously, the expression levels of p-AMPK and p-ACC in HepG2 cells and the liver tissue of MSG mice were increased, while the expression levels of PEPCK and p-GS were decreased after treatment with linarin. Conclusion: Insulin resistance could be ameliorated by linarin in type 2 diabetes, and its mechanism may be related to AMPK signaling pathway.

Preparation and pharmacokinetics in vivo of linarin solid dispersion and liposome / 中草药·英文版

Objective: The current investigation aimed to determine the appropriate dosage form by comparing solid dispersion and liposome to achieve the purpose of improving the solubility and bioavailability of linarin. Methods: Linarin solid dispersion (LSD) and linarin liposome (LL) were developed via the solvent method and the thin film hydration method respectively. The Transwell chamber model of Caco-2 cells was established to evaluate the absorption of drug. The pharmacokinetics of linarin, LSD and LL in rats after ig administration were carried out by high performance liquid chromatography (HPLC) method. Results: The solubility of LSD and LL was severally 3.29 times and 3.09 times than that of linarin. The permeation coefficients of LSD and LL were greater than 10

Study on HPLC fingerprints of Biyuanjing capsules / 药学实践杂志

Objective To establish a HPLC fingerprints of Biyuanjing capsules. Methods The column was Agilent SB-C18(4.6mm×250 mm, 5 µm). The mobile phase was acetonitrile-water with gradient elution at a flow rate of 1.0 ml/min. The detection wavelength was 210 nm. The detection time was 80 min. Results The HPLC fingerprints of Biyuanjing capsules were established. Twenty common peaks were confirmed, of which, 15 peaks were belonging to each crude drug and 5 peaks were identified as chemical components. The overall similarity of the fingerprints of 10 batches of samples was above 90% comparing with the control. Conclusion This method can be used for the quality control of Biyuanjing capsules.

Correlations between content of linarin in Chrysanthemum indicum and climatic factors in habitats / 中国中药杂志

Chrysanthemi Indici Flos(CIF), the capitulum of Chrysanthemum indicum, is widely used in proprietary Chinese medicine and daily chemical products. At present, CIF is mainly produced from wild resources and rarely cultivated. This study aims to reveal the correlations between linarin content in CIF and climatic factors in different habitats, and provide a theoretical basis for suitable zoning and rational production of medicinal materials. The content of linarin in CIF was determined by HPLC. Grey relational analysis and Pearson correlation analysis were carried out for linarin content with climatic factors. The results showed that the content of linarin in CIF was significantly different among different habitats. The grey relational degrees of climatic factors with linarin content was in an order of average annual precipitation>annual average sunshine hours>annual average temperature>longitude>annual frost-free period>latitude>altitude. Longitude, annual average temperature and average annual precipitation had significantly positive correlations with the content of linarin in CIF, whereas latitude and altitude showed negative correlations with it. The annual frost-free period and annual average sunshine hours had no significant correlation with the content of linarin in CIF. The content of linarin in CIF varied significantly in different habitats. High longitude, low latitude, low altitude, high annual average temperature and high annual average precipitation could be used as indicators for the habitats of high-quality Ch. indicum. This study provides a reference for selecting suitable producing areas of Ch. indicum and establishing artificial cultivation system.

Simultaneous determination of five active components in Fangshu Qingre mixture by RP-HPLC / 药学实践杂志

Objective To establish a method for simultaneous determination of chlorogenic acid, luteolin-7-O-β-D-glucoside,3,5-dicaffeoylquinic acid, linarin and pogostone in Fangshu Qingre mixture by RP-HPLC. Methods ZORBAX-SB-C18 column(4.6 mm×250 mm, 5 μm) was used as the chromatographic column.The mobile phase was 0.2% formic acid water solution(A) and acetonitrile solution (B)with a gradient elution mode. The flow rate was 1.0 ml/min.The detective wavelength was 327 nm. The column temperature was 30 ℃. Results There were good linear relationships in the determination of chlorogenic acid, luteolin-7-O-β-D-glucoside, 3,5-dicaffeoylquinic acid, linarin and pogostone (r≥0.999 6), with the average recovery rate of 102.03%(1.63%), 102.38%(1.51%), 102.39%(1.23%), 103.14%(1.87%) and 104.01%(2.33%). Conclusion The method was simple and stable with a good reproducibility, which could be used as a quality control method for active compotents in Fangshu Qingre mixture.

Preparation and Quality Evaluation of Standard Decoction of Chrysanthemi Indici Flos Based on Traditional Decoction Process / 中国实验方剂学杂志

Objective:To study the dry extract rate,determination and transfer rate of maker compounds,fingerprint and others of standard decoction of Chrysanthemi Indici Flos and provide basic data for the preparation of this standard decoction and its dispensing granules by establishing 15 batches of standard decoction of Chrysanthemi Indici Flos from 5 different places. Method:The standard decoction of Chrysanthemi Indici Flos was prepared based on the traditional decoction process,the content of linarin was determined by UPLC-DAD,the transfer rate of this composition was calculated,the fingerprint was drawn,the extract powder was prepared by vacuum drying,and the dry extract rate was calculated. Result:The concentration of linarin in 15 batches of standard decoction of Chrysanthemi Indici Flos was 0.19-0.74 g·L-1,the transfer rate of linarin was 21.95%-66.23%,its average transfer rate was 37.12% with RSD of 11.8%,the pH value was 5.1-5.5,the range of dry extract rate was 24.7%-32.5%,the average dry extract rate was 27.87% with RSD of 2.4%.There were 9 major common peaks in the fingerprint and 2 peaks(No. 2 and No. 9) were confirmed,such as chlorogenic acid and linarin. Conclusion:The preparation method in this research conforms to the traditional decoction method and is stable and feasible.It can be used for the preparation and quality evaluation of standard decoction of Chrysanthemi Indici Flos.

Effects of application of N, P, and K on traits and active ingredients of Chrysanthemum indicum / 中草药

Objective To explore the effect of N, P, and K fertilizers on the growth of Chrysanthemum indicum and accumulation of active components. Methods This research used different levels and ratios of nitrogen, phosphorus and potassium through orthogonal experiment; The flower of C. indicum traits were observed in the harvest period; The content of the dried flower active ingredient was measured using HPLC. Results The result showed the noticeable differences among the 14 groups in the number of capitulum, the diameter of capitulum and tubiform floret, yield per plant, total flavonoid content and linarin content as well as fertilizer efficiency of different mineral elements. Appropriate increase of N, K concentration and low P concentration was beneficial to the growth and development of C. indicum and the output of dried flowers. Low concentration of N, P, K promoted the cotent of linarin and total flavonoids; The content of linarin treated with different combination all accorded with the standard of Chinese Phamaeopoeia. Conclusion Compared comprehensively, it was concluded that the optimal proportion of nitrogen, phosphorus and potassium in the growth of chrysanthemum inflorescence15:0.1:3. Flavonoid and linarin content reached the highest point without fertilization.

Chemical constituents from the aerial parts of saururus chinensis() / 中国药学杂志

OBJECTIVE: To study the chemical constituents from the aerial parts of Saururus chinensis. METHODS: The compounds were extracted by supercritical CO2 extraction and isolated by various chromatographic methods, such as silica gel, MCI and pre-HPLC. The structures were elucidated by physico-chemical constants and spectroscopic methods. RESULTS: Fourteen compounds were isolated and identified as N-p-trans-coumaroyltyramine (1), linarin (2), blumenol A (3), mannitol (4), 5,7-dyhydroxyl-3,4′-bimethoxyflavonoids (5), 6,7-dimethoxy-4-hydroxy-1-naphthalene formic acid (6)3-hydroxy-4-methoxy benzoic acid (7), (2R,3R)-2,3-dihydro-2-(4-hydroxy-3-methoxyphenyl)-7-methoxy-3-methylbenzofuran-5-aldehyde (8), (6S,7E)-6-hydroxy-4,7-megastigmadien-3,9-dione (9), veratric acid (10), p-hydroxybenzaldehyde (11), syringaldehyde (12), 3,4,5-trimethoxybenzoic acid (13) and physcion (14). CONCLUSION: Compounds 1-13 are isolated from this plant for the first time.

Establishment of the QAMS method for the determination of rutin， linarin，luteolin and apigenin in Herba Cirsii / 国际药学研究杂志

Objective: To establish a method for the quantitative analysis of multi-components by single marker (QAMS)to determine the content of rutin,linarin,luteolin and apigenin in Herba Cirsii. Methods: Rutin was selected as the internal reference substance,the relative correction factors(RCF)of the other 3 components were established re- spectively,and then the content of each component was calculated according to respective RCF. At the same time,the external standard method was used to determine the content of all four components in Herba Cirsii,the differences be- tween calculated values and measured values were compared,and the method validation was performed. Results: No significant difference was observed between the calculated values and measured ones according to the results from ten batches of Herba Cirsii samples. Conclusion: The validated QAMS method is proved to be of good precision,reproduc- ibility,and reliability which is suitable and feasible for the quality control of Herba Cirsii

Determination of 3 components in Xinjiang Ziziphora bungeana by HPLC / 中国中药杂志

This study aimed to develop an HPLC method for simultaneous determination of the 3 components in Ziziphora bungeana. The optimum HPLC condition was as follows：ZORBAX SB-C₁₈ column（4.6 mm×250 mm, 5.0 μm）with gradient elution of methanol (A)-0.2% glacial acetic acid (B), detection wavelength 340 nm,column temperature 30 °C, flow rate 1 mL·min⁻¹. There were good linearity between peak areas and injection quantity of caffeic acid, rosmarinic acid, and linarin in the range of 2-40 mg·L⁻¹（=0.999 9）, 3-60 mg·L⁻¹（=1）, 7-140 mg·L⁻¹（r=0.999 9）, respectively. The average recoveries were 100.9%（RSD 1.3%）,98.25%（RSD 2.0%）,and 98.73%（RSD 1.5%）, respectively. The HPLC method was stable and accurate, which could be used to detect caffeic acid,rosmarinic acid, and linarin in Ziziphora bungeana.

Simultaneous Determination of Five Active Constituents in Cirsii Herba Dispensing Granules by HPLC-DAD / 中国药师

Objective: To develop an HPLC-DAD method for the simultaneous determination of five active constituents (chlorogen-ic acid, baicalin, rutin, linarin and apigenin) in Cirsii Herba dispensing granules. Methods: The chromatographic separation was per-formed on a Thermo Hypersil BDS C18column (250 mm×4. 6 mm,5 μm) with gradient elution by 0. 1% phosphoric acid-acetonitrile as the mobile phase at the flow rate of 1. 0 ml·min-1. The detection wavelength was set at 326 nm,and the column temperature was maintained at 35 ℃. Results: Chlorogenic acid, baicalin, rutin, linarin and apigenin were linear within the range of 0. 072-1. 446 μg (r=0.999 7), 0.043-0.864 μg(r =0.999 5), 0.094 2-1.884 μg (r =0.999 7), 0.150-3.000 μg (r =0.999 8) and 0.043-0. 856 μg (r=0. 999 6), respectively. The recovery was 97. 67% , 98. 19% , 97. 92% ,98. 12% and 98. 02% , and the RSDs was 0. 82% , 0. 92% , 1. 25% , 0. 98% and 1. 17% , respectively. Conclusion: The method is accurate, convenient and reproducible in the quality control of multicomponents in Cirsii Herba dispensing granules.

Quality standard of Chrysanthemum indicum / 中草药

Objective To improve the quality standard of Flos Chrysanthemum Indicum from diffrent origins by analyzing on linarin and cumambrin A qualitatively and quantitatively. Methods Qualitative analysis of linarin and cumambrin A was carried out by thin layer chromatography (TLC); Content determination of linarin and cumambrin A was using high performance liquid chromatography (HPLC) on YMC C18 column (250 mm × 4.6 mm, 5 μm); The mobile phase was a mixture of acetonitrile-0.05% phosphoric acid solution; The elution mode was gradient system (0-18 min, 25%-26% A; 18-26 min, 26%-32% A; 26-33 min, 32%-34% A; 33-35 min, 34%-40% A; 35-65 min, 40%-50% A); The flow rate was 0.8 mL/min; The detection wavelengths were 203 nm and 340 nm; The column temperature was 35 ℃. Results Linarin and cumambrin A by TLC was obvious. The concents of linarin and cumambrin A in Flos Chrysanthemum Indicum from diffrent origins were different. The concents of linarin and cumambrin A from Xinyang were the highest (6.53% and 0.81% respectively). Conclusion It is the first time to establish a method to evaluate different components in Flos Chrysanthemum Indicum by TLC and HPLC. The method is simple, accurate and reproducible, which can effectively improve the existing quality standard of Flos Chrysanthemum Indicum. The result also showed Linarin and cumambrin A could reflect the quality of Flos Chrysanthemum Indicum.

Study on chemical fingerprint of Wuwei Xiaoduyin Oral Liquid and simultaneous determination of six characteristic components / 中草药

Objective: To establish the HPLC chemical fingerprints of Wuwei Xiaoduyin Oral Liquid (WXOL) and simultaneous determination method for six major characteristic components in this herbal preparation, i.e. chlorgenic acid, luteoloside, luteolin, linarin, caffeic acid, and esculetin. Methods: Both chemical fingerprint analysis and quantitative determination were implemented by Agilent 1260 high performance liquid chromatography system with an Agilent 5 TC-C18 column (250 mm × 4.6 mm, 5 μm). A gradient elution program, with the mobile phase of 0.5% glacial acetic acid aqueous solution (A) and methanol (B), was employed as following: 0—10 min, 10%—32% B; 10—20 min, 32% B; 20—25 min, 32%—46% B; 25—31 min, 46%—48% B; 31—41 min, 48%—80% B at the flow rate of 1.0 mL/min. The detection wavelength and column temperature were set at 320 nm and 30 ℃, respectively. Additionally, fingerprint similarity of 10 batches of WXOL was evaluated by software “Similarity Evaluation System for Chromatographic Fingerprint of TCM” published by GPC (Version 2012). The amounts of six characteristic components in WXOL were also simultaneously determined. Results: The common fingerprint pattern derived from 10 batches of samples was obtained with the similarity over 0.98 and 17 common peaks defined. Meanwhile, some common peaks were identified via peak pattern match with standard substances, showing that the peak No.5, No.7, No.8, No.12, No.16, and No.17 was chlorgenic acid, esculetin, caffeic acid, luteoloside, linarin, and lutelin, respectively. Chlorgenic acid content range of 54.038 3—105.551 1 μg/mL, esculetin content range of 4.122 1—31.359 9 μg/mL, caffeic acid content range of 2.413 0—4.420 7 μg/mL, luteoloside content range of 4.042 8—11.312 8 μg/mL, linarin content range of 3.866 3—46.271 9 μg/mL, and luteolin content range of 0.990 8—2.126 8 μg/mL. In combination with the non-significant difference of multiple characteristic components content among 10 batches of samples, the quality of home-made WXOL would be stable. Conclusion: A novel quality control method, which is HPLC fingerprint in combination with simultaneous quantitative analysis of multiple components, was established in this study, with high repeatability and reliability. Therefore, this method provides an applicable approach for the quality control of WXOL.

Simultaneous Determination of Scutellarin and Linarin in Zhuang Medicine Scoparia dulcis by HPLC / 中国药房

OBJECTIVE:To establish a method for the simultaneous determination of scutellarin and linarin in Scoparia dulcis. METHODS:HPLC was performed on the column of Phenomenex C18 with mobile phase of acetonitrile-0.05% phosphoric acid (gradient elution)at a flow rate of 1.0 ml/min,the detection wavelength was 330 nm,column temperature was 30 ℃,and the in-jection volume was 10 μl. RESULTS:The linear range was 0.015 8-0.316 8 mg/ml for scutellarin(r=0.999 9)and 0.000 5-0.010 4 mg/ml for linarin (r=0.999 7);RSDs of precision,stability and reproducibility tests were lower than 3%;recoveries were 99.56%-102.38%(RSD=1.07%,n=6)and 95.14%-98.29%(RSD=1.24%,n=6),respectively. CONCLUSIONS:The method is simple with good stability and reprodicibility,and can be used for the simultaneous determination of scutellarin and linarin in S. du-lcis.

Study on balance of process of alcohol precipitation of ganmaoling granules / 中国中药杂志

Ganmaoling granule is the first brand of domestic cold medicine sales, but its preparation method and process control parameters are relatively rough. Therefore it is urgent to upgrade the technologies of large varieties of traditional Chinese medicine (TCM). This paper focused on the balance between the remove of impurity and the retention of linarin during the process of alcohol precipitation of Ganmaoling granules. The effects of four factors on the process were investigated via single factor experiments. The results showed that the precipitating period, the initial ethanol concentration and the final ethanol concentration had a great effect on retention of linarin while the initial density of the extract has not. Similarly, the initial ethanol concentration, the final ethanol concentration and the initial extract density have a great effect on the yield of dry extract while the time of alcohol precipitation has not. The parameters of alcohol precipitation of Ganmaoling granules were optimized as 16 h of precipitating period, 95% ethanol as the initial reagent, 70% of the final ethanol concentration, and 1.10 of the initial extract density.

Modeling of thermal degradation of linarin during concentration process / 中国中药杂志

Ganmaoling granule, with annual sale of over one billion yuan, is the first brand of domestic cold medicine sales. As the only traditonal Chinese medicine（TCM） quality control indicator of Ganmaoling granule, linarin is thermally unstable. Its content will be changed significantly during the production process, which would then affect the quality of the finished product. In this paper, the law of degradation of linarin was investigated. The experimental results showed that degradation reaction of linarin belongs to the first reaction characteristics. The effective methods to reduce the loss of linarin would be realized fortunately by strictly controlling the heating temperature or shortening the heating time.

Simultaneous Determination of Four Components in Shenju Lotion by HPLC / 中国药房

OBJECTIVE:To establish a method for simultaneous determination of matrine,linarin,berberine hydrochloride and osthole in Shenju lotion. METHODS:HPLC was performed on the column of Dionex C18 with mobile phase of acetonitrle-0.1% triethylamine(pH was adjusted to 7.2 with phosphoric)(gradient elution)at a flow rate of 1.0 ml/min,detection wavelength was 210 nm,column temperature was 30 ℃,and injection volume was 20 μl. RESULTS:The linear range was 1.400-8.400 μg for matrine (r=0.999 4),0.152-0.912 μg for linarin (r=0.999 6),0.248-1.488 μg for berberine hydrochloride (r=0.999 9) and 0.128-0.768 μg for osthole (r=0.999 4);limit of quantitation was 84.85,3.15,15.03,7.76 ng,limit of detection was 24.13, 0.89,4.27,2.20 ng;RSDs of precision,stability and reproducibility tests were lower than 2%;recoveries were 96.29%-99.06%(RSD=1.10%,n=9),96.49%-98.27%(RSD=0.69%,n=9),99.26%-101.13%(RSD=0.62%,n=9) and 96.27%-98.64%(RSD=0.89%,n=9). CONCLUSIONS:The method is simple,accurate and reproducible,and can be used for the simultaneous determination of matrine,linarin,berberine hydrochloride and osthole in Shenju lotion.

Simultaneous Determination of Seven Active Constituents in Different Parts of Dandrantherna Morifolium ( Ramat. ) Tzvel. by HPLC-DAD / 中国药师

Objective:To develop an HPLC-DAD method for the simultaneous determination of seven active constituents chloro-genic acid, 3,5-dicaffeoylquinic acid,linarin, luteolin-7-O-β-D-glucoside, apigenin-7-O-β-D-glucoside , luteolin and acacetin in the different parts of Dandrantherna morifolium( Ramat. ) Tzvel. Methods:The chromatographic separation was performed on a Diamonsil C18 column (200 mm × 4. 6 mm,5 μm) with 0. 2% phosphoric acid-acetonitrile as the mobile phase with gradient elution at the flow rate of 1 ml·min-1 . The detection wavelength was set at 348 nm,and the column temperature was maintained at 30 ℃. Results:Chlorogenic acid, 3,5-dicaffeoylquinic acid, linarin, luteolin-7-O-β-D-glucoside, apigenin-7-O-β-D-glucoside , luteolin and acacetin was linear within the range of 0.014-0.560 μg(r =0.999 5) ,0.031-1.260 μg(r =0.999 6), 0.022-0.900 μg(r =0.999 8), 0. 036-1. 440 μg(r=0. 999 6),0. 029-1. 160 μg (r=0. 999 7) , 0. 005 5-0. 220 0 μg (r=0. 999 8) and 0. 006 5-0. 260 0 μg (r=0. 999 9), respectively. The average recovery was 97. 91%, 97. 91%, 97. 55%, 96. 43%, 96. 44%, 97. 07% and 97. 48%, and RSDs was 1. 24%, 1. 29%, 1. 48%, 1. 83%, 0. 67%, 1. 88% and 1. 80%, respectively. Conclusion: The method is convenient, accurate and reproducible in the quality control of the multicomponents in different parts of Dandrantherna morifolium( Ramat. ) Tzvel.

Effects of Cynaroside, Cynarin and Linarin on Secretion, Production and Gene Expression of Airway MUC5AC Mucin in NCI-H292 Cells

In this study, we investigated whether cynaroside, cynarin and linarin derived from Chrysanthemum indicum L. affect the secretion, production and gene expression of MUC5AC mucin in airway epithelial cells. Confluent NCI-H292 cells were pretreated with cynaroside, cynarin or linarin for 30 min and then stimulated with PMA (phorbol 12-myristate 13-acetate) for 24 h. The MUC5AC mucin gene expression, mucin protein production and secretion were measured by RT-PCR and ELISA, respectively. Effect of linarin on EGF (epidermal growth factor) - or TNF-alpha (tumor necrosis factor-alpha)-induced MUC5AC mucin gene expression and mucin protein production was also examined. The results were as follows: (1) Cynaroside and cynarin did not significantly affect PMA-induced MUC5AC mucin secretion from NCI-H292 cells. However, linarin decreased MUC5AC mucin secretion; (2) Cynaroside did not affect PMA-induced MUC5AC mucin production and gene expresion from NCI-H292 cells. However, cynarin and linarin inhibited the production and gene expression of MUC5AC mucin; (3) Linarin also inhibited the production and gene expression of MUC5AC mucin induced by EGF- or TNF-alpha from NCI-H292 cells. These results suggest that linarin can regulate the gene expression, production and secretion of mucin, by directly acting on airway epithelial cells.