Application of active material of sea cucumber as medication or health products
A technology for active substances and health products, which is applied in the application field of sea cucumber active substances as medicines or health products, and can solve problems such as no sea cucumber extracts.
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Embodiment 1
[0020] Extraction of sea cucumber active substances.
[0021] Fresh sea cucumber, gutted, washed, broken into 1cm pieces 3 The left and right pieces are homogenized by adding 5-15 times of water.
[0022] The homogenate was subjected to pepsin (the amount of enzyme was 0.4% of the wet weight of sea cucumber, pH 2.0-2.5, temperature 50°C, time 5h), trypsin (the amount of enzyme was 0.45% of the wet weight of sea cucumber, pH 8.0-8.5, Temperature 50°C, time 5h) enzymolysis, after enzymolysis, boiling to inactivate the enzyme activity, centrifugation (10000r / min, 10min) to take the supernatant.
[0023] The supernatant was precipitated with alcohol, centrifuged (6000r / min, 15min) to obtain the precipitate, then dissolved in distilled water, separated through a DEAE cellulose column, and the absorption peak was detected at 210nm, and the active component was collected (using the activity of nerve cells as an index), and lyophilized.
[0024] The freeze-dried sea cucumber active...
Embodiment 2
[0027] Proliferative effect of sea cucumber active substances on neural stem cells.
[0028] Wistar rats at 13.5-14 days pregnant were sacrificed by cervical dislocation.
[0029] Isolate and take out the embryos under aseptic conditions, and separate them in a sterile petri dish under a dissecting microscope to obtain nerve cells. Place them in a carbon dioxide incubator at 37°C and 5% carbon dioxide saturated humidity for 48 hours before performing group experiments.
[0030] After the nerve cells were cultured in the 96-well plate for 48 hours, 5 mg / ml of the sea cucumber extract prepared in Example 1 were added to make the final concentrations of the sea cucumber extract respectively 100 μg / ml, 200 μg / ml, 300 μg / ml, and 400 μg / ml, a normal control group was set up, cultured at 37°C, 5% carbon dioxide saturated humidity for 48 hours, and then MTT was measured.
[0031] Dose / (μg·ml-1)
[0032] It can be seen from Table 1 that the extract of sea cucumber has obvi...
Embodiment 3
[0034] Protective effect of sea cucumber active substances on oxidative damage of nerve cells.
[0035] Pregnant 13.5-14 day Wistar rats were taken and sacrificed by cervical dislocation.
[0036] Isolate and take out the embryos under aseptic conditions, and separate them in a sterile petri dish under a dissecting microscope to obtain nerve cells. Place them in a carbon dioxide incubator at 37°C and 5% carbon dioxide saturated humidity for 48 hours before performing group experiments.
[0037] After the nerve cells were cultured in the 96-well plate for 48 hours, 5 mg / ml of the sea cucumber extract prepared in Example 1 were added to make the final concentrations of the sea cucumber extract respectively 100 μg / ml, 200 μg / ml, 300 μg / ml, and 400 μg / ml, a normal control group was set up.
[0038] After culturing for 12 hours, glutamic acid culture solution was added to each well so that the final concentration of glutamic acid was 2 mmol / ml. After culturing for 36 hours at 37...
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