Composition for relieving pain and preventing or alleviating stress-involved disease, comprising cannabis extract as active ingredient
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[0077]Preparation of cannabis Extract
[0078]Cannabis including leaves and flowers was washed thoroughly with running water and then air-dried completely. The dried cannabis was pulverized with a mixer and then prepared as a powder. After 50% ethanol as an extraction solvent was added to the powder sample at a ratio of 1:10 (w:v) and then the powder sample was completely immersed, extraction was performed repeatedly three times at 80° C. under reflux for each 3 hours. The liquid extract was filtered with a Whatman No. 2 filter paper. A cannabis extract (CE) was prepared by concentrating the filtrate at 60° C. under reduced pressure.
[0079]2. Preparation of Other Natural Extracts
[0080]An Erigeron annuus (L.) Pers extract (EE), a Ceratonia siliqua (L.) Taub. extract (IE), and a Stellaria media extract (SE) were prepared using a method which is the same as the preparation method of the cannabis extract (CE).
[0081]3. Preparation of Complex Extract
[0082]A complex extract was formed by mi...
experimental example 2
in
[0087]Cyclooxygenase (COX)-2 Inhibition Ability
[0088]The cannabis extract (CE), which was a sample used for the cyclooxygenase (COX)-2 enzyme inhibition ability measurement experiment, was dissolved in a DMSO solution and used. First, after the COX-2 enzyme, heme, and an inhibitory sample were put into a test tube containing a buffer for a reaction (the control group contained the same amount of DMSO instead of the inhibitory sample), the mixture was allowed to react at 37° C. for 10 minutes. After the reaction was completed for 10 minutes, 10 μL of a substrate (arachidonic acid) was added thereto, and the reaction was further performed for 2 minutes. When the reaction was completed, the reaction was terminated by adding 50 μL of a 1M HCl solution thereto, and a reduction reaction was caused by adding tartaric acid. After each test tube reaction solution was diluted by 2000 fold and 4000 fold, 50 μL of the reaction solution was each added to a 96-well plate (well plate). After 50 ...
experimental example 3
and Improving Stress
[0101]4-week-old ICR male mice {SAMTAKO Bio Korea, Korea} were subjected to an adaptation time of 1 week, and then bred in a space where lighting is turned on and off repeatedly in a unit of 12 hours, and an indoor temperature of 18 to 23° C. and a humidity of 60% were maintained. As a feed, a solid feed was supplied, and there was no restriction on the feed or grade other than the process of inducing stress.
[0102]After the complex extracts (MX1 to MX6) in the Preparation Example were dissolved in distilled water, the resulting solutions were orally administered (40 mg / kg) continuously once a day at 14:00 to 15:00 for one month. The experimental group consisted of a total of three groups, which are a control group to which only stress was applied, a group to which the complex extracts (MX1 to MX6) were administered, and a normal control group to which neither stress nor extract was given. Five experimental animals were used for each experimental group, and stress...
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