Cockroach polypeptide effective part for treating ulcerative colitis as well as preparation method and application thereof
A technology for ulcerative colitis and effective parts, which is applied in the field of medicine and can solve problems such as poor product activity
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Embodiment 1
[0058] Add 500g of fresh cockroach body, add petroleum ether 5L, soak overnight in a multifunctional extraction tank, heat and reflux at 50°C for extraction, extract 3 times, each time for 3h, filter, discard the filtrate, dry the residue at 50°C for 6h, and dry the residue About 150g. Put 150g of the dried residue in a multifunctional extraction tank, add 1.5L of water, heat and extract at 90°C, extract 3 times, each time for 2h, filter, and concentrate the filtrate to a density of 1.14-1.16g / mL (60°C). Add 95% ethanol, adjust the alcohol content to 50%, let stand at 15°C for 48 hours, filter, recover ethanol, concentrate to a relative density of 1.28-1.30g / mL (60°C), freeze-dry to obtain the crude extract of cockroaches PA1.
[0059] Dry 180g of cockroach body, add petroleum ether 5L, soak in a multifunctional extraction tank overnight, heat and reflux at 50°C for extraction, extract 3 times, each time for 3h, filter, discard the filtrate, dry the residue at 50°C for 6h, an...
Embodiment 2
[0064] Add 500g of fresh cockroaches, add petroleum ether 5L, soak overnight in a multifunctional extraction tank, heat and reflux at 50°C for extraction, extract 3 times, each time for 3h, filter, discard the filtrate, dry the residue at 50°C for 6h, and the dried residue is about 150g . Put 150g of the dried residue in a multifunctional extraction tank, add 1.5L of water, heat and extract at 90°C, extract 3 times, each time for 2h, filter, and concentrate the filtrate to a density of 1.14-1.16g / mL (60°C). Add 95% ethanol to adjust the alcohol content, let stand at 15°C for 48h, filter, recover ethanol, concentrate to a relative density of 1.28-1.30g / mL (60°C), freeze-dry to obtain the crude extract of cockroaches.
[0065] Table 2 Yield and activity index evaluation of cockroach bold substance under different alcohol contents
[0066]
[0067] According to the results in Table 2, with the increase of alcohol content, the total peptide mass and the 24h relative migration ...
Embodiment 3
[0069] Dissolve the crude extract of cockroaches in sterile double-distilled water, adjust the pH with 5% ammonia water, place in a dialysis bag with a molecular weight of 500Da, stir magnetically at 300rpm, and dialyze 4 times, each time for 5h; then transfer the solution in the dialysis bag Put it into a 5KDa dialysis bag, stir magnetically at 300rpm, continue dialysis, dialysis 4 times, 5h each time; collect the dialysate, filter, combine the filtrates, concentrate at 50°C, and freeze-dry to obtain the effective part of the cockroach polypeptide.
[0070] Table 3 Yield and Activity Index Evaluation of Effective Parts of Cockroach Polypeptides at Different pH
[0071]
[0072] According to the results in Table 3, with the increase of pH, the yield, the quality of the polypeptide, and the relative migration ability of Caco-2 cells for 24 hours all increased, and the in vitro anti-inflammatory activity did not change significantly. Considering the cost control of large-scale...
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