Anti-inflammatory Effect of lanceolata Root Extracts Hee-Ock Boo1*, Jeong-Hun Park1, Hag-Hyun Kim2, Soo-Jeong Kwon2, Moon-Soon Lee3 and Sun-Hee Woo4 1WELLPHYTO Co. Ltd., BI Center, GIST, Gwangju 61005, Republic of 2Faculty of Food Nutri.&Cookery, Woosong Information College, Daejeon 34606, Republic of Korea 3Dept. of Industrial Sci.&Tech., Chungbuk National University, Cheongju 28644, Republic of Korea 4Dept. of Crop Science, Chungbuk National University, Cheongju 28644, Republic of Korea Introduction Codonopsis lanceolata is a perennial belonging to the family , and is grown commercially in East Asia. The roots of C. lanceolata have been used as a crude tonic drug in Korea, and an edible plant mainly contain triterpenoid saponins including codonolaside Ⅰ-V, lancemaside A-G. The saponins obtained from the plant have exhibited anti-inflammatory effects such as bronchitis and cough, insomnia, and hypomnesia. C. lanceolata is well known to affect various pharmacological effects on human health, and its consumption is increasing. Recently, plant and plant-derived products have treated a part of the healthcare system by applying the bioactive phytochemicals. This study was conducted to investigate candidate material as an anti-inflammation agent from root extracts of C. lanceolata. Materials and Methods Roots of C. lanceolata plant were freeze-dried and then ground into a fine powder. The powder was stored at -20˚C for further experiments. The powder was immersed in 30% ethanol, and the filtrate was collected for three times with constant stirring of the mixture at every 24 hrs interval of a 72 hrs total collection period. The extract was then concentrated under reduced pressure at 45˚C using a vacuum rotary evaporator. The concentrated extract was partitioned between hexane and water. The aqueous layer further fractionated with methyl chloride, ethyl acetate, and butyl alcohol. Four solvent fractions (hexane, methyl chloride, ethyl acetate and butyl alcohol) were collected and concentrated using vacuum rotary evaporator. Anti-inflammatory activities were evaluated on the several inflammation-related factors such as nitric oxide (NO) and the release of proteinflammatory cytokine [tumor necrosis factor-alpha (TNF-α), Interleukin-6 (IL-6) and Interleukin-1 beta (IL-1β)] in lipopolysaccharide (LPS)- treated RAW 264.7 cells. Results and Discussion This study was executed to evaluate candidate material as an anti-inflammation agent from C. lanceolata. The inhibitory effect of TNF-α, IL-6, IL-1β secretion and NO production in LPS--treated RAW 264.7 cells was found highly dependent on the ethanol concentration. The anti-inflammatory effects of the extract fractions were higher in the ethyl acetate and the n-hexane fraction than in the other fractions. Most of the fractions were thought to have anti-inflammatory activity. The result from this investigation suggests that the extract of C. lanceolata has potential anti-inflammatory efficacy in vitro cells, and then could be an addition to medicine for anti-inflammatory modulation.

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0 0 0.0 0.00 LPS H.W 30% 50% 70% 100% Hot water EtOH 30% EtOH 50% EtOH 70% EtOH 100% LPS H.W 30% 50% 70% 100% LPS H.W 30% 50% 70% 100% LPS (1ug/ml) LPS (1ug/ml) LPS (1ug/ml) ______CE ( 5ug/ml) CE ( 5ug/ml) CE ( 5ug/ml) Fig. Effect of Codonopsis lanceolata according to the percentage of EtOH solvent on cell viability, and NO production, and LPS-induced TNF-α, IL-1β and IL-6 in RAW 264.7 cells.

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0 0.0 0.0 0.00 LPS n-H MC EA n-BA DW LPS n-H MC EA n-BA DW LPS n-H MC EA n-BA DW LPS n-H MC EA n-BA DW LPS (1ug/ml) LPS (1ug/ml) LPS (1ug/ml) LPS (1ug/ml) ______CE ( 5ug/ml) CE ( 5ug/ml) CE ( 5ug/ml) CE ( 5ug/ml) Fig. Inhibitory effect by different solvent fractions of Codonopsis lanceolata on NO production, and LPS-induced TNF-α, IL-1β and IL-6 in RAW 264.7 cells. n-H: n-hexane, MC: Methylene chloride, EA: Ethyl acetate, n-BA: n- Butyl alcohol, DW: distilled water Acknowledgement This research was supported by High Value-added Food Technology Development Program (No. 112076-03-3- HD030) of IPET, Ministry of Agriculture, Food and Rural Affairs, Republic of Korea. *Corresponding Author E-mail: [email protected]