阅读说明：本技术 东北赤杨提取物及制备方法、应用 (Alnus japonica extract, preparation method and application thereof ) 是由 张昌浩 邬楠 金莉莉 尹雄杰 于 2020-08-10 设计创作，主要内容包括：本发明公开了一种东北赤杨提取物,是按下列方法制备：取东北赤杨干燥的皮,称重；以料液比1g：30mL的比例加入65%~95%乙醇溶液,浸泡24h；在恒温55℃~70℃加热回流提取,提取3次,每次提取时间3h,趁热过滤,舍去滤渣,合并滤液；将滤液减压浓缩至浸膏,放入烘箱干燥；取烘干后浸膏,即得。本发明还公开了一种东北赤杨提取物的制备方法、应用。本发明具有工艺简便,提取速度快,提取率高且活性强等优点,具有较好的提取效果。提取物具有肝保护作用,可用于制备肝保护药物等研究。(The invention discloses a northeast Alnus japonica extract, which is prepared by the following method: weighing dried bark of Alnus japonica in northeast China; 1g of liquid-material ratio: adding 65-95% ethanol solution into 30mL of the mixture, and soaking for 24 hours; heating and refluxing for extraction at the constant temperature of 55-70 ℃, extracting for 3 times, wherein the extraction time is 3h each time, filtering while hot, discarding filter residues, and combining filtrates; concentrating the filtrate under reduced pressure to obtain extract, and drying in oven; and (5) drying the extract to obtain the extract. The invention also discloses a preparation method and application of the Alnus japonica extract. The invention has the advantages of simple and convenient process, high extraction speed, high extraction rate, strong activity and the like, and has better extraction effect. The extract has liver protecting effect, and can be used for preparing liver protecting medicine.)

1. A Buxus japonica extract is prepared by the following steps:

(1) weighing dried bark of Alnus japonica in northeast China;

(2) 1g of liquid-material ratio: adding 65-95% ethanol solution into 30mL of the mixture, and soaking for 24 hours;

(3) heating and refluxing for extraction at the constant temperature of 55-70 ℃, extracting for 3 times, wherein the extraction time is 3h each time, filtering while hot, discarding filter residues, and combining filtrates;

(4) concentrating the filtrate under reduced pressure to obtain extract, and drying in oven;

(5) and (5) drying the extract to obtain the extract.

2. The extract of Alnus japonica Thunb as claimed in claim 1, wherein the dried bark of Alnus japonica Thunb is extracted by refluxing with 95% ethanol at a constant temperature of 65 ℃.

3. A preparation method of an Alnus japonica extract comprises the following steps:

(1) weighing dried bark of Alnus japonica in northeast China;

(2) 1g of liquid-material ratio: adding 65-95% ethanol solution into 30mL of the mixture, and soaking for 24 hours;

(3) heating and refluxing for extraction at the constant temperature of 55-70 ℃, extracting for 3 times, wherein the extraction time is 3h each time, filtering while hot, discarding filter residues, and combining filtrates;

(4) concentrating the filtrate under reduced pressure to obtain extract, and drying in oven;

(5) and (5) drying the extract to obtain the extract.

4. Use of the extract of Alnus japonica Thunb as claimed in claim 1 for liver protection.

Technical Field

The invention belongs to the technical field of plant extraction and pharmacological activity of medicinal plants, and particularly relates to a northeast Alnus japonica extract as well as a preparation method and application thereof.

Background

The northeast Alnus japonica belongs to Alnus plant of Betulaceae. Alnus (Alnus)Alnus Mill) The plant is deciduous tree or shrub. The genus is more than 40 species, and is distributed in Asia, Africa, Europe and North America. 7 Chinese-made Chinese herbs1 variety, distributed in northeast, east, south, China and southwest. The inferior classification included short alder, alder group and single-sequence group. Alder plants generally have anti-inflammatory and antioxidant effects, and are widely used in oriental folks as a medicine for treating fever, hemorrhage, diarrhea, and alcohol. It has been reported that many bioactive natural ingredients are isolated from this genus of plants, including diarylheptanes, polyphenols, flavones, terpenes, steroids, and the like. The alder plant can be used for medicine without toxic and side effects, and has good development prospect and research potential.

The liver is responsible for the management of qi, blood and water circulation in the body. Emotional disorder, sleep, diet, even drugs, etc., all affect the function of the liver to smooth flow of qi, resulting in liver damage. If the treatment is not timely carried out, more serious physical diseases may appear, and in recent years, liver protection is found in some medicinal plants, so that the liver protection tea is worthy of intensive study.

Disclosure of Invention

The invention provides a high-efficiency extracted Alnus japonica extract and a preparation method thereof, and application of the Alnus japonica extract in liver protection, aiming at the prior art, thereby providing a foundation for subsequent drug development.

In order to realize the purpose, the provided technical scheme is as follows:

a Buxus japonica extract is prepared by the following steps:

(1) weighing dried bark of Alnus japonica in northeast China;

(2) 1g of liquid-material ratio: adding 65-95% ethanol solution into 30mL of the mixture, and soaking for 24 hours;

(3) heating and refluxing for extraction at the constant temperature of 55-70 ℃, extracting for 3 times, wherein the extraction time is 3h each time, filtering while hot, discarding filter residues, and combining filtrates;

(4) concentrating the filtrate under reduced pressure to obtain extract, and drying in oven;

(5) and (5) drying the extract to obtain the extract.

Further, the dried bark of the Alnus japonica in northeast China is extracted by 95% ethanol in a reflux manner at a constant temperature of 65 ℃.

A preparation method of an Alnus japonica extract comprises the following steps:

(1) weighing dried bark of Alnus japonica in northeast China;

(2) 1g of liquid-material ratio: adding 65-95% ethanol solution into 30mL of the mixture, and soaking for 24 hours;

(3) heating and refluxing for extraction at the constant temperature of 55-70 ℃, extracting for 3 times, wherein the extraction time is 3h each time, filtering while hot, discarding filter residues, and combining filtrates;

(4) concentrating the filtrate under reduced pressure to obtain extract, and drying in oven;

(5) and (5) drying the extract to obtain the extract.

The application of the above-mentioned Alnus japonica extract in liver protection is provided.

The invention provides an optimal extraction process of the northeast Alnus japonica, which adopts an ethanol solution to carry out heating reflux extraction, can obtain higher extraction effect in a shorter time and effectively improve the utilization rate of the northeast Alnus japonica; in the experiment, a liver injury model is manufactured for a mouse, LPS/D-GaIN mixed solution is injected into the abdominal cavity, and then the liver of the liver injury model mouse, the liver of the northeast Alnus japonica dosing group and the liver of the blank group are compared to confirm the liver protection effect of the northeast Alnus japonica.

The extraction process disclosed by the invention has the advantages of simple process, high extraction speed, high extraction rate, strong activity and the like, and has a better extraction effect. The extract has liver protecting effect, and can be used for preparing liver protecting medicine.

Drawings

FIG. 1 shows liver function test values of each experimental group (A: blank group; B: model group; C: low dose group; D: high dose group) in the present embodiment.

FIG. 2 is an observed view of HE stained liver sections of each experimental group (A: blank group; B: model group; C: low dose group; D: high dose group) according to an embodiment of the present invention.

Detailed Description

A Buxus japonica extract is prepared by the following steps:

(1) weighing dried bark of Alnus japonica in northeast China;

(2) 1g of liquid-material ratio: adding 65-95% ethanol solution into 30mL of the mixture, and soaking for 24 hours;

(3) heating and refluxing for extraction at the constant temperature of 55-70 ℃, extracting for 3 times, wherein the extraction time is 3h each time, filtering while hot, discarding filter residues, and combining filtrates;

(4) concentrating the filtrate under reduced pressure to obtain extract, and drying in oven;

(5) and weighing the dried extract, and calculating the extraction rate.

The invention relates to an extraction process of Alnus japonica in northeast China, which comprises the following 3 key conditions: ethanol, reflux extraction temperature and reflux extraction time. The following single factor experiment was performed for three aspects.

Effect of ethanol concentration

Table 1 shows the results of a single-factor experiment of ethanol concentration at an extraction temperature of 65 ℃ and an extraction time of 3 h: the ethanol concentration is preferably 95%.

TABLE 1 influence of ethanol concentration on extraction yield

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Influence of extraction temperature

Table 2 shows the results of a single-factor experiment of the extraction temperature at an ethanol concentration of 95% and an extraction time of 3 h: the extraction temperature is preferably 65 deg.C.

TABLE 2 Effect of extraction temperature on extraction yield

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Influence of extraction time

Table 3 shows the results of a single factor experiment of extraction time at an ethanol concentration of 95% and an extraction temperature of 65 ℃: the extraction time is preferably 3 h.

TABLE 3 Effect of extraction time on extraction yield

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Orthogonal test of extraction process conditions of Alnus japonica

TABLE 4 results of orthogonal experiments

。

The above experiments show that: through single factor experiment and orthogonal experiment, the extraction rate under different extraction process conditions is obtained, and the optimal process conditions are that the ethanol concentration is 95%, the extraction temperature is 65 ℃, and the extraction time is 3 hours.

The application of the above-mentioned Alnus japonica extract in liver protection, in particular to the research on the liver protection effect of Alnus japonica in northeast, comprises the following steps:

(1) male ICR mice weighing 23-28g were divided into 4 groups (blank, model, high and low), five per group; alternating light and shade for 12h, and cutting off grains and water;

(2) preparing LPS (15 mug/Kg) and D-GaIN (600 mg/Kg) into a mixed solution for later use;

(3) preparing 300mg/Kg and 100mg/Kg of the Alnus japonica extractum for later use;

(4) respectively gavage 300mg/Kg and 100mg/Kg of northeast Alnus japonica to high and low dose mice; the other groups are infused with physiological saline with the same amount as the stomach;

(5) after 30min, injecting LPS/D-GaIN mixed solution into abdominal cavity of other groups except blank group, and injecting equal amount of normal saline into blank group;

(6) after 6h, serum aspartate Aminotransferase (AST) and alanine Aminotransferase (ALT) are detected by blood collection of orbital venous plexus of mice; liver indexes MDA and SOD are detected and statistical analysis is carried out.

(7) And (3) taking the livers of the mice of each group to carry out wax block embedding, and preparing tissue sections to carry out HE staining observation.

The study on the liver protection effect of the northeast Alnus japonica comprises the success of model building and whether the liver injury of mice is inhibited by different doses of the northeast Alnus japonica. The experiment was carried out in two ways.

The mouse is subjected to the intraperitoneal injection of the LPS/D-GaIN mixed solution, the mouse is observed for 48 hours, the mouse does not die, the liver of the model group mouse is taken out and compared with the liver taken out of the normal group mouse, the color of the liver is darker, a certain number of white nodules appear, the liver cells of the model group under an electron microscope deform, and a large number of vacuoles exist in the cells, so that the liver of the group of mice is proved to have certain damage, and the model is proved to be successfully manufactured.

The mice adopt high and low doses for intragastric administration, the liver damage degree of a high and low dose administration group is detected to be improved compared with a model group, and the liver damage degree is expressed as AST reduction, ALT reduction, MDA reduction and SOD rise, the improvement effect of the high dose group is better, all experimental data are expressed by x +/-s, single-factor variance analysis is carried out by using GraphPadPrism5.0 statistical software, and P <0.05 shows that the difference has statistical significance; after taking out the 4 groups of livers, HE staining comparison is carried out, compared with a model group, the damage degree of the livers of the administration group is reduced, the effect of the high-dose group is more obvious, and the populus deltoids have certain liver protection effect and have no toxic or side effect.

The liver function test values of the experimental groups (A: blank group; B: model group; C: low dose group; D: high dose group) were as shown in FIG. 1, and the HE stained liver section observation pattern was as shown in FIG. 2, which indicated that: experimental research shows that the populus deltoides has liver protection effect and no toxic or side effect, and can be used for preparation of subsequent liver protection medicines and other researches.

The foregoing is only a preferred embodiment of the present invention and the present invention is not limited to the above examples. Modifications, changes or substitutions by one of ordinary skill in the art within the spirit of the present invention should be included within the scope of the present invention.