阅读说明：本技术 一种促进成骨细胞增殖治疗骨质疏松的制备方案 (Preparation scheme for promoting osteoblast proliferation and treating osteoporosis ) 是由 王澄宇 于 2021-10-25 设计创作，主要内容包括：本发明公开了一种促进成骨细胞增殖治疗骨质疏松的制备方案,包括如下制备步骤：选取罗非鱼鳞,在罗非鱼鳞中加入其重量5～7倍、质量浓度为1～5g/L的酸溶液,均匀浸泡40～150分钟；然后加热至55～60℃并保温搅拌10～60分钟；过滤除掉滤渣,在滤液中按6000～8000U/g胶原基质添加蛋白酶,并调整pH在6.5～7.5范围内、温度50～55℃下反应7～8h；然后升温至沸腾并保持1～2min,即得酶解液。本发明通过将可增加骨密度的组合物制备颗粒,相对于现有的重要服用方法而言,方便服用,便于运输,质量更加稳定,剂量准确,同时,罗非鱼鳞促骨形成肽具有较好的促进成骨细胞增殖活性,可促进成骨细胞ALP表达,有利于成骨细胞的分化和矿化。(The invention discloses a preparation scheme for promoting osteoblast proliferation and treating osteoporosis, which comprises the following preparation steps: selecting tilapia mossambica scales, adding an acid solution which is 5-7 times of the tilapia mossambica scales in weight and has a mass concentration of 1-5 g/L, and uniformly soaking for 40-150 minutes; then heating to 55-60 ℃ and stirring for 10-60 minutes under heat preservation; filtering to remove filter residues, adding protease into the filtrate according to 6000-8000U/g of collagen matrix, and adjusting the pH value to be 6.5-7.5 and reacting for 7-8 h at the temperature of 50-55 ℃; and then heating to boil and keeping for 1-2 min to obtain the enzymatic hydrolysate. Compared with the existing important taking method, the preparation method has the advantages that the preparation method is convenient to take and transport, the quality is more stable, the dosage is accurate, and meanwhile, the tilapia mossambica scale bone formation promoting peptide has good activity of promoting osteoblast proliferation, can promote the ALP expression of osteoblasts and is beneficial to differentiation and mineralization of osteoblasts.)

1. A preparation scheme for promoting osteoblast proliferation and treating osteoporosis is characterized by comprising the following preparation steps:

s1, selecting tilapia mossambica scales, adding an acid solution which is 5-7 times of the tilapia mossambica scales in weight and has a mass concentration of 1-5 g/L, and uniformly soaking for 40-150 minutes;

s2, heating to 55-60 ℃, and stirring for 10-60 minutes under heat preservation;

s3, filtering to remove filter residues, adding protease into the filtrate according to 6000-8000U/g of collagen matrix, and adjusting the pH value to be 6.5-7.5 and reacting at 50-55 ℃ for 7-8 h; then heating to boil and keeping for 1-2 min to obtain an enzymolysis liquid;

s4, filtering the enzymatic hydrolysate while the enzymatic hydrolysate is hot to obtain clear liquid, and then concentrating and drying the clear liquid to obtain collagen peptide;

s5 selecting radix Puerariae, adding saline water into radix Puerariae, stirring, adding 2 parts of salt into per 100 parts of radix Puerariae, parching, taking out, and cooling;

s6 adding 6 times of water into parched radix Puerariae, decocting for 2 hr, filtering to obtain filtrate, concentrating the filtrate, oven drying at low temperature to obtain radix Puerariae extract, and grinding into powder;

s7 mixing collagen peptide and radix Puerariae extract, adding calcium carbonate, and mixing.

2. The preparation method of claim 1, wherein the mixture obtained in step S7 is hermetically packaged.

3. The preparation method of promoting osteoblast proliferation for treating osteoporosis as claimed in claim 1, wherein the acid solution of step S1 is hydrochloric acid, acetic acid, lactic acid, or phosphoric acid.

4. The preparation method of claim 1, wherein the filtrate obtained in step S6 is concentrated under reduced pressure at 80-85 ℃ to obtain a thick paste with a relative density of 1.20-1.25.

5. The preparation method of claim 1, wherein the step S4 is to perform coarse filtration on the obtained enzymolysis solution, add activated carbon to the filtrate after the coarse filtration, keep the temperature at 40-50 ℃ for 20-25 min, and finally perform membrane filtration to obtain a clear solution.

6. The preparation scheme of claim 5, wherein the pore size of the membrane filtration is 0.1 μm or 0.22 μm.

7. The preparation method of claim 1, wherein the step S4 is implemented by drying the supernatant through vacuum freeze drying or low-temperature spray drying.

Technical Field

The invention relates to the technical field of improving bone mineral density, in particular to a preparation scheme for promoting osteoblast proliferation and treating osteoporosis.

Background

Osteoporosis is a common and frequent disease in China, and has a very serious influence on the health of middle-aged and elderly people, especially postmenopausal women. Osteoporosis can cause various complications such as fracture, thoracic deformity and the like, and not only can lead patients to suffer from pain, but also can lead the patients and families to bear heavy life and economic pressure. It is expected that the number of patients with osteoporosis and low bone mass will increase to 2.8 hundred million by 2020 in our country. National large-scale epidemiological survey also shows that the total disease rate of the osteoporosis in China is 12.4 percent, and the disease proportion of the old people is over 50 percent. Osteoporosis has seriously threatened the health of middle-aged and elderly people.

The patent with the application number of CN201510689506.2 discloses a preparation method of bioactive peptide, which aims to solve the problem that the prior method for obtaining protein peptide for promoting osteoblast proliferation activity from lactoferrin is lacked. The preparation method comprises the following steps: firstly, mixing lactoferrin with deionized water, and adjusting the pH value; secondly, adding pepsin into the lactoferrin solution, and carrying out water bath reaction; thirdly, separating and purifying by adopting a cation exchange column after filtering, and then dialyzing and freeze-drying; fourthly, purifying by adopting a gel filtration chromatographic column after filtering; and fifthly, drying to obtain the lactoferrin peptide. The lactoferrin peptide prepared by using bovine lactoferrin as a raw material has a good effect of promoting the proliferation of osteoblasts. The prepared lactoferrin peptide can be applied to health-care food or functional food. However, the reasonable utilization rate of the processing by-products is low at present, and the reasonable utilization is inconvenient.

Disclosure of Invention

Based on the technical problems in the background art, the invention provides a preparation scheme for promoting osteoblast proliferation and treating osteoporosis.

The invention provides a preparation scheme for promoting osteoblast proliferation and treating osteoporosis, which comprises the following preparation steps:

s1, selecting tilapia mossambica scales, adding an acid solution which is 5-7 times of the tilapia mossambica scales in weight and has a mass concentration of 1-5 g/L, and uniformly soaking for 40-150 minutes;

s2, heating to 55-60 ℃, and stirring for 10-60 minutes under heat preservation;

s3, filtering to remove filter residues, adding protease into the filtrate according to 6000-8000U/g of collagen matrix, and adjusting the pH value to be 6.5-7.5 and reacting at 50-55 ℃ for 7-8 h; then heating to boil and keeping for 1-2 min to obtain an enzymolysis liquid;

s4, filtering the enzymatic hydrolysate while the enzymatic hydrolysate is hot to obtain clear liquid, and then concentrating and drying the clear liquid to obtain collagen peptide;

s5 selecting radix Puerariae, adding saline water into radix Puerariae, stirring, adding 2 parts of salt into per 100 parts of radix Puerariae, parching, taking out, and cooling;

s6 adding 6 times of water into parched radix Puerariae, decocting for 2 hr, filtering to obtain filtrate, concentrating the filtrate, oven drying at low temperature to obtain radix Puerariae extract, and grinding into powder;

s7 mixing collagen peptide and radix Puerariae extract, adding calcium carbonate, and mixing.

Preferably, the mixture obtained in step S7 is sealed and packaged.

Preferably, the acid solution of step S1 is hydrochloric acid, acetic acid, lactic acid, or phosphoric acid.

Preferably, the filtrate obtained in step S6 is concentrated under reduced pressure at 80-85 ℃ to form a thick paste with a relative density of 1.20-1.25.

Preferably, in the step S4, the obtained enzymolysis solution is firstly subjected to coarse filtration, then activated carbon is added to the filtrate after the coarse filtration, the temperature is kept at 40-50 ℃ for 20-25 min, and finally, a clear solution is obtained through membrane filtration.

Preferably, the pore size of the membrane filtration is 0.1 μm or 0.22 μm.

Preferably, the step S4 is to perform a drying process on the clear solution by vacuum freeze drying or low-temperature spray drying.

Compared with the existing important taking method, the preparation method has the advantages that the preparation method is convenient to take and transport, the quality is more stable, the dosage is accurate, meanwhile, tilapia mossambica scale bone formation promoting peptide has good activity of promoting osteoblast proliferation, can promote the ALP expression of osteoblasts and is beneficial to differentiation and mineralization of the osteoblasts.

Drawings

FIG. 1 is a schematic flow chart of a preparation scheme for treating osteoporosis by promoting osteoblast proliferation according to the present invention.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments.

Referring to fig. 1, a preparation scheme for promoting osteoblast proliferation to treat osteoporosis comprises the following preparation steps:

s1, selecting tilapia mossambica scales, adding an acid solution which is 5-7 times of the tilapia mossambica scales in weight and has a mass concentration of 1-5 g/L, and uniformly soaking for 40-150 minutes;

s2, heating to 55-60 ℃, and stirring for 10-60 minutes under heat preservation;

s3, filtering to remove filter residues, adding protease into the filtrate according to 6000-8000U/g of collagen matrix, and adjusting the pH value to be 6.5-7.5 and reacting at 50-55 ℃ for 7-8 h; then heating to boil and keeping for 1-2 min to obtain an enzymolysis liquid;

s4, filtering the enzymatic hydrolysate while the enzymatic hydrolysate is hot to obtain clear liquid, and then concentrating and drying the clear liquid to obtain collagen peptide;

s5 selecting radix Puerariae, adding saline water into radix Puerariae, stirring, adding 2 parts of salt into per 100 parts of radix Puerariae, parching, taking out, and cooling;

s6 adding 6 times of water into parched radix Puerariae, decocting for 2 hr, filtering to obtain filtrate, concentrating the filtrate, oven drying at low temperature to obtain radix Puerariae extract, and grinding into powder;

s7 mixing collagen peptide and radix Puerariae extract, adding calcium carbonate, and mixing.

In the present invention, the mixture obtained in step S7 is hermetically packaged.

In the present invention, the acid solution of step S1 is hydrochloric acid, acetic acid, lactic acid, or phosphoric acid.

In the invention, the filtrate obtained in step S6 is concentrated under reduced pressure at 80-85 ℃ to form thick paste with the relative density of 1.20-1.25.

In the invention, step S4 is to firstly carry out coarse filtration on the obtained enzymolysis liquid, then add active carbon into the filtrate after the coarse filtration, keep the temperature at 40-50 ℃ for 20-25 min, and finally obtain clear liquid by membrane filtration.

In the present invention, the pore size of the membrane filtration is 0.1 μm or 0.22. mu.m.

In the present invention, the step S4 is to perform a drying process on the clear solution by vacuum freeze drying or low-temperature spray drying.

The invention comprises the following steps: selecting tilapia mossambica scales, adding an acid solution which is 5-7 times of the tilapia mossambica scales in weight and has a mass concentration of 1-5 g/L, and uniformly soaking for 40-150 minutes; then heating to 55-60 ℃ and stirring for 10-60 minutes under heat preservation; filtering to remove filter residues, adding protease into the filtrate according to 6000-8000U/g of collagen matrix, and adjusting the pH value to be 6.5-7.5 and reacting for 7-8 h at the temperature of 50-55 ℃; then heating to boil and keeping for 1-2 min to obtain an enzymolysis liquid; filtering the enzymatic hydrolysate while the enzymatic hydrolysate is hot to obtain clear liquid, and then concentrating and drying the clear liquid to obtain collagen peptide; selecting radix Puerariae, adding saline water into radix Puerariae, stirring, adding 2 parts of salt into 100 parts of radix Puerariae, parching thoroughly, taking out, and cooling; adding 6 times of water by weight into fried dried radix puerariae, decocting for 2 hours, filtering to obtain filtrate, concentrating the filtrate, drying at low temperature to obtain radix puerariae extract, and grinding into powder; mixing collagen peptide and radix Puerariae extract, adding calcium carbonate, and mixing.

The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be considered to be within the technical scope of the present invention, and the technical solutions and the inventive concepts thereof according to the present invention should be equivalent or changed within the scope of the present invention.