Pharmaceutical composition for the prevention or treatment of alopecia comprising eremochloa ophiuroides extract or fractions thereof as an active ingredient
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example 1
ON OF CENTIPEDE GRASS EXTRACT
[0111]The centipede grass seeds were purchased from Fukukaen Nursery and Blub Co. Ltd, Japan, which grew in the field at Advanced Radiation Technology Institute, Korea Atomic Energy Research Institute, Jeongeup, Korea. Leaves were collected from the grown centipede grass. The obtained leaves were stored at −80° C. 7 kg of the centipede grass leaves were mixed with 225 l of 80% methanol was added thereto 3 times, and the mixture was grinded in a mixer. Precipitation was induced at room temperature for 3 days. The obtained extract was concentrated under reduced pressure at 50° C. As a result, 502.5 g of a centipede grass methanol extract was prepared.
example 2
ON OF CENTIPEDE GRASS FRACTION
[0112]The centipede grass methanol extract obtained in Example 1 was fractionated using organic solvents according to the polarity. The methanol extract concentrated under reduced pressure was suspended in 3 l of water added with 10% methanol. First, 3 l of hexane (N-hexane), the low-polar solvent was added thereto 5 times. The fraction was concentrated under reduced pressure and dried. As a result, 44.0311 g of a hexane fraction was obtained. Then, 3 l of ethyl acetate was added to the water layer 5 times, followed by fractionation. The fraction was concentrated under reduced pressure and dried. As a result, 28.1328 g of an ethyl acetate fraction was obtained. Then, 3 l of butanol was added to the water layer 5 times, followed by fractionation. The fraction was concentrated under reduced pressure and dried. As a result, 111.1267 g of a butanol fraction and 213.371 g of a water-soluble fraction were obtained.
experimental example 1
ELL PROLIFERATION ACTIVITY
[0113]The effect of the ethyl acetate and butanol fractions of the centipede grass extract on the skin cell (HaCaT) proliferation was investigated by MTT assay.
[0114]Particularly, HaCaT cells were diluted in the medium supplemented with 10% FBS, which was distributed in a 24 well plate at the density of 50,000 cells / well, followed by culture for 24 hours. Upon completion of the culture, the medium was replaced with the medium supplemented with 1% FBS to minimize the effect of other ingredients included in the serum, followed by culture for 24 hours. Thereafter, the medium was added or not added with the centipede grass fraction at the concentration of 100 ng / ml, 1 μg / ml, or 10 μg / ml, and the cells were further cultured. The medium of the cultured cells was replaced with the medium supplemented with 0.5 mg / ml of MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] (Amresco, USA), followed by culture at 37° C. 3 hours later, MTT reagent was clea...
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