The present study is to investigate which kinds of solvent extracts

The present study is to investigate which kinds of solvent extracts of Inulae Flos inhibit the chemokine productions in HaCaT cell and whether the inhibitory capacity of Inulae Flos is related with constitutional compounds. fraction and inhibited the secretion of chemokines dose-dependently more than the other compounds. Luteolin also represented dose-dependent inhibition on chemokine productions although it was contained at lower levels in 70% methanol extract and solvent fractions. These results suggest that the inhibitory effects of Inulae Flos on chemokine production in HaCaT Calcipotriol manufacturer cell could be related with constituent compounds contained, especially 1, 5-dicaffeoylquinic acid and luteolin. 1. Introduction Inulae Flos, the inflorescence of or (Asteraceae), has demonstrated therapeutic efficacy by Calcipotriol manufacturer reducing phlegm, advertising the dissipation of pathological water, redirecting the qi downward, and preventing vomiting. The therapeutic effectiveness of Inulae Flos offers prompted its use in the treatment of symptoms such as the build up of phlegm and fluids clogging up the lungs, vomiting, hiccough, belching, and cough with excessive expectoration of phlegm [1]. Recent pharmacological studies of Inulae Flos have shown hepatoprotective [2], immunoregulatory [3], antidiabetic [4], hypolipidemic [4], anticancer [5], antiinflammatory [6], antioxidant, and neuroprotective properties [7] when it was evaluated as water or organic solvent components of the whole natural medicine. Its pharmacological activity has been associated not only with the whole natural medicinal draw out but also with compounds extracted from your natural medicine. Although crude components of a single natural medicine or natural formula can show striking biological effects, their mechanisms cannot be fully established because countless compounds are contained in even a solitary natural medicine. Most studies of the biological effects or mechanisms of herbal medicines concentrate on the main compound of the natural medicine. Compounds isolated from Inulae Flos have shown pharmacological activities, such as iNOS inhibition by 1-O-acetyl-4R,6S-britannilactone [8], the antitumour effects of sesquiterpenelactones [9], the antidiabetic effects of polysaccharides [10], the antioxidative effects of flavonoids [11], and the inhibition of NO production by sesquiterpenes [12]. The compounds contained in natural medicine can be recognized with analytical techniques, and the predominant compound is definitely often thought to be strongly associated with the biological effect. The chemical Calcipotriol manufacturer compounds in Inulae Flos were analysed with high-performance liquid chromatography-ultraviolet detection (HPLC-UV), as reported in earlier papers, and the constructions of the flavonoids and sesquiterpenes were identified [13, 14]. In the present study, we extracted Inulae Flos with different solvent compositions then further partitioned the components to determine the constituent having the predominant biological effect. The concentrations of eight compounds of Inulae Flos were quantified in extract by different solvent compositions and solvent fractions to determine the relationships between the inhibitory effect of Inulae Flos and its constituent compounds on chemokine productions in HaCaT cell. 2. Materials and Method 2.1. Reagents and Flower Materials HPLC-grade methanol, ethanol, acetonitrile, and water were purchased from J. T. Baker Inc. (Phillipsburg, NJ, USA). Caffeic acid (99%) and chlorogenic acid (99%) were purchased from Acros Organics (NJ, USA). Rutin LIMK2 (95%), quercetin (98%), and luteolin (99%) were from Sigma-Aldrich (St Louis, MO, USA). 6-Methoxy-luteolin and kaempferol-3-O-glucoside were purchased from ChromaDex (Irvine, CA, USA) and 1,5-dicaffeoylquinic acid (99.2%) from Chengdu Biopurify Phytochemicals (Chengdu, China). The chemical constructions of the standard compounds were classified as phenylpropanoids and flavonoids, as demonstrated in Number 1. Inulae Flos was from local market of natural medicine (Kwangmyungdang Medicinal Natural herbs, Ulsan, Republic of Korea). A voucher specimen (ST2011-13) was deposited in the Basic Natural Medicine Research Group of the Korea Institute of Oriental Medicine. Open in a separate window Number 1 Chemical constructions of the components of Inulae Flos. Chlorogenic acid (1), caffeic acid (2), 1,5-dicaffeoylquinic acid (3), rutin (4), kaempferol-3-O-glucoside (5), quercetin (6), luteolin (7), and 6-methoxy-luteolin (8). 2.2. Extraction of the Natural Medicine The dried aerial portion of Inulae Flos (1.0?g) was pulverized through a 60 mesh sieve and extracted with 100?mL of 70% (v/v) methanol, 70% (v/v) ethanol, 100% methanol, 100% ethanol, and deionized water for 60?min with sonication, respectively. Each draw out was filtered through a SmartPor GHP syringe filter (Woongi Technology, Seoul, Korea) before it was injected into the HPLC apparatus. The remaining components were filtered through a paper filter (Advantec, Japan) and concentrated having a rotary evaporator under vacuum for biological testing. The yields of the components were 10.05% in 70% methanol, 11.93% in 70% ethanol, 7.29% in 100% methanol, 4.04% in 100% ethanol, and 12.51% in deionized water. 2.3. Partitioning of the.