Their possible cytotoxic effect was examined using MTT assay. using Western blot analysis. Results: Nitric oxide level produced by LPS-primed macrophages was significantly decreased with all prepared fractions in a dose-dependent manner. Saturated sesquiterpene lactones-rich species (and plants may be associated with the inhibition of pro-inflammatory signaling pathways. However, these effects could be dependent on the type of their sesquiterpene content. These findings also introduce new species cultivated in Iran as a useful anti-inflammatory agents. belongs the family Asteraceae, which are comprises about 550 species distributed in Europe, Asia, and North America (1). The leaves are very aromatic, alternate, often with long, narrow segments, usually grayish or silvery, hairy. The flower heads are greenish or brownish, without rise. The fruits are seeded, usually flattened or ribbed, without a pappus. The genus in Iran has 34 species which two of them are endemic (2). Some species including L. (Afsantin), L. (Shih), L. (Qaysum Zakar) and L. (Tarkhun) have been used extensively in folk medicine to alleviate several ailments (3). species exhibit a wide range of leishmanicidal (4), cytotoxic (5-7), anti-microbial (8) and anti-oxidant activities (8). More importantly, the anti-inflammatory effects of species have been reported (9-11). A number of bioactive compounds including acetylenes, coumarins, terpenes, monoterpenes, monoterpene lactones, sesquiterpenes, sesquiterpene lactones, flavonoids, dipeptides, phenolics, coumarin, ethers, esters, esterols and polysaccharides are commonly found in members of the genus (12). In recent years, several sesquiterpenes derived from including artemisinin and dihydroarteannuin (13), artemisolide and eupatilin (14), scoparone and capillarisin (15), scopoletin (16) have received special attention due to their pharmacological activity on inflammatory processes and other illnesses.? Nitric oxide (NO) is mainly synthesized by inducible NO synthase (iNOS) which is largely involved in the pathophysiology of many inflammatory diseases (17). Another key enzyme in inflammatory responses is cyclooxygenase-2 (COX-2) which is responsible for prostaglandin E2 (PGE2) production (18). Several inflammatory stimuli such as bacterial lipopolysachharide (LPS) could activate iNOS and COX-2 expression. Various agents could serve as an important therapeutic target in the treatment of various inflammation-based pathologies (19, 20). Many medicines commonly used for the treatment of inflammatory diseases could impart various adverse side-effects (21). Numerous researches have focused on herbal cures with lower adverse effects and improved efficacy. Taking into consideration the above facts, present study aimed to evaluate and compare the anti-inflammatory effects of sesquiterpene fractions isolated from various species through effects on the production of NO and PGE2 as well as on the expression of iNOS and COX-2 by LPS-primed J774A.1 macrophages. To our knowledge, no other study has been carried out to compare the anti-inflammatory effect of species in regard to their sesquiterpene contents. Aside from this, essentially nothing is known regarding the potential anti-inflammatory property of some species tested here. Materials and Methods lipopolysaccharide (LPS, serotype 0111:B4), sodium nitrite, species (Table 1) collected from different regions of Khorasan Province, Iran and identified by Dr V Mozaffarian (Research Institute of Forest and Rangelands, Ministry of Jahad Keshavarzi, Iran). Voucher specimens were deposited in the Herbarium of National Botanical Garden of Iran (TARI). The shade dried and powdered plant samples were preserved for further experimentations. Table 1 Tested species Open in a separate window Open in a separate window Table 2 1H-NMR key data for the sesquiterpene lactone fractions of tested species Open in a separate window Open in a separate window Sesquiterpene fractions were prepared using Herz-H?genauer technique (22). The chlorophyll and common phenolic were removed by lead-()-acetate precipitation and preparing crude sesquiterpene samples for further chromatographic and spectral investigations. Dried and ground plant materials (20 g) were soaked in dichloromethane (DCM; 100 ml) overnight. The slurry products were then filtered and evaporated production species were 92.490.86, 90.402.60, 90.002.15, 91.540.56, most effectively reduced NO release by LPS-activated J774A.1 macrophages. Open in a separate window Figure 1 Effect of sesquiterpene lactone fractions of different species including A) on LPS-stimulated J774A.1 macrophages production of NO. Cells were treated for a total of 24 hr with LPS alone or after an initial 2-hr treatment.To our knowledge, no other study has been carried out to compare the anti-inflammatory effect of species BI 2536 in regard to their sesquiterpene contents. was significantly decreased with all prepared fractions in a dose-dependent manner. Saturated sesquiterpene lactones-rich species (and plants may be associated with the inhibition of pro-inflammatory signaling pathways. However, these effects could be dependent on the type of their sesquiterpene content. These findings also introduce new species cultivated in Iran as a useful anti-inflammatory agents. belongs the family SERK1 Asteraceae, which are comprises about 550 species distributed in Europe, Asia, and North America (1). The leaves are very aromatic, alternate, often with long, narrow segments, usually grayish or silvery, hairy. The flower heads are greenish or brownish, without rise. The fruits are seeded, usually flattened or ribbed, without a pappus. The genus in Iran has 34 species which two of them are endemic (2). Some species including L. (Afsantin), L. (Shih), L. (Qaysum Zakar) and L. (Tarkhun) have been used extensively in folk medicine to alleviate several ailments (3). species exhibit a wide range of leishmanicidal (4), cytotoxic (5-7), anti-microbial (8) and anti-oxidant activities (8). More importantly, the anti-inflammatory effects of species have been reported (9-11). A number of bioactive compounds including acetylenes, coumarins, terpenes, monoterpenes, monoterpene lactones, sesquiterpenes, sesquiterpene lactones, flavonoids, dipeptides, BI 2536 phenolics, coumarin, ethers, esters, esterols and polysaccharides are commonly found in members of the genus (12). In recent years, several sesquiterpenes derived from including artemisinin and dihydroarteannuin (13), artemisolide and eupatilin (14), scoparone and capillarisin (15), scopoletin (16) have received special attention due to their pharmacological activity on inflammatory processes and other illnesses.? Nitric oxide (NO) is mainly synthesized by inducible NO synthase (iNOS) which is largely involved in the pathophysiology of many inflammatory diseases (17). Another key enzyme in inflammatory responses is cyclooxygenase-2 (COX-2) which is responsible for prostaglandin E2 (PGE2) production (18). Several inflammatory stimuli such as bacterial lipopolysachharide (LPS) could activate iNOS and COX-2 expression. Various agents could serve as an important therapeutic target in the treatment of various inflammation-based pathologies (19, 20). Many medicines commonly used for the treatment of inflammatory diseases could impart various adverse side-effects (21). Several researches have focused on natural remedies with lower adverse effects and improved effectiveness. Taking into consideration the above details, present study targeted to evaluate and compare the anti-inflammatory effects of sesquiterpene fractions isolated from numerous varieties through effects within the production of NO and PGE2 as well as within the manifestation of iNOS and COX-2 by LPS-primed J774A.1 macrophages. To our knowledge, no additional study has been carried out to compare the anti-inflammatory effect of varieties in regard to their sesquiterpene material. Aside from this, essentially nothing is known concerning the potential anti-inflammatory BI 2536 house of some varieties tested here. Materials and Methods lipopolysaccharide (LPS, serotype 0111:B4), sodium nitrite, varieties (Table 1) collected from different regions of Khorasan Province, Iran and recognized by Dr V Mozaffarian (Study Institute of Forest and Rangelands, Ministry of Jahad Keshavarzi, Iran). Voucher specimens were deposited in the Herbarium of National Botanical Garden of Iran (TARI). The color dried and powdered flower samples were maintained for further experimentations. Table 1 Tested varieties Open in a separate window Open in a separate window Table 2 1H-NMR important data for the sesquiterpene lactone fractions of tested varieties Open in a separate window Open in a separate windowpane Sesquiterpene fractions were prepared using Herz-H?genauer technique (22). The chlorophyll and common phenolic were eliminated by lead-()-acetate precipitation and preparing crude sesquiterpene samples for further chromatographic and spectral investigations. Dried and ground flower materials (20 g) were soaked in dichloromethane (DCM; 100 ml).