CN105616462B - Moringa oleifera extract for promoting intestinal growth and preparation method and application thereof - Google Patents
Moringa oleifera extract for promoting intestinal growth and preparation method and application thereof Download PDFInfo
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- CN105616462B CN105616462B CN201610140138.0A CN201610140138A CN105616462B CN 105616462 B CN105616462 B CN 105616462B CN 201610140138 A CN201610140138 A CN 201610140138A CN 105616462 B CN105616462 B CN 105616462B
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- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
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- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/55—Liquid-liquid separation; Phase separation
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Abstract
The invention discloses a moringa oleifera extract for promoting intestinal growth and a preparation method and application thereof. The preparation method of the moringa extract comprises the following steps: extracting moringa oleifera with ethanol, extracting the extract with ethyl acetate, and layering to obtain ethyl acetate extract; concentrating the ethyl acetate extract, loading the ethyl acetate extract to a silica gel chromatographic column, eluting by using a petroleum ether-acetone system, then eluting by using methanol, and collecting methanol eluent; concentrating the methanol eluent, loading the concentrated methanol eluent on macroporous adsorption resin, eluting with ethanol, collecting ethanol eluent, concentrating, and drying to obtain the moringa oleifera extract. The invention has the advantages that: can effectively promote intestinal canal and intestinal villus growth.
Description
Technical Field
The invention belongs to the technical field of medicines, and particularly relates to a moringa oleifera extract for promoting intestinal growth, and a preparation method and application thereof.
Background
The term "intestinal Failure" appears in the literature in the 50 s of the 20 th century and is used up to now, but has no complete meaning and has definite monitoring parameters like other organs. Before the 80's of the 20 th century, the main function of the intestinal tract was considered to be digestion and absorption. Therefore, the monitoring index of the intestinal function is mainly the degree of digestion and absorption. After the 80 s, the importance of intestinal mucosal barrier function was recognized in clinical work, and intestinal bacterial translocation (enterobacterial translocation 2) has had a significant role in causing pathophysiological changes in critically ill patients. Intestinal mucosal barrier function has been considered as another important function of the intestinal tract, particularly late-stage concurrent infections in many critically ill patients, although possibly originating from other routes, and is impaired by factors such as hypoxia, ischemia, etc., and intestinal bacterial translocation is the major source. Intestinal bacterial translocation has been clearly demonstrated in animal experiments, and bacteria can be observed to enter the lymphatic or portal systems either directly through the intestinal mucosal cell tight junctions or through the cells.
Short bowel syndrome is one of the major causes of intestinal failure, and it refers to a group of clinical syndromes characterized mainly by diarrhea, weight loss and malnutrition due to extensive excision of the small intestine. Survival by long-term TPN is achieved when adults retain at least 60-80cm (15% to 20% of normal length) of the small intestine. Survival is supported by parenteral nutrition when the newborn infant has a residual small intestine (no ileocaecal valve) of at least 40cm or a ileocaecal valve of at least 10-20 cm. The quality of life and outcome of the patient depends on the remaining length of the small intestine and its functional status. During the course of increased intestinal adaptation following intestinal resection, patients are generally encouraged to take food orally unless they are clinically stimulated to adapt to the progress, unless intestinal failure and its severity. This is due to adaptation changes depending on food, bile and pancreatic secretions in the intestinal lumen; the intestinal peripheral nutrition support such as TPN has no effect on the adaptation and growth of intestinal tracts, but can cause a series of complications such as gallstone, liver failure and the like. Therefore, it is a safe and effective treatment to administer food or medicine effective for promoting intestinal growth during the oral ingestion treatment stage of patients with short bowel syndrome.
Moringa (Moringa oleifera Lam.) is a Moringa plant of the Moringaceae family, belonging to the genus small arbor plant, evergreen or semideciduous, originally produced in India, also known as Qizhu, drumstick, and perennial deciduous tree, widely planted in the tropical and subtropical regions of Asia and Africa, with about 13 Moringa species throughout the world, mainly distributed in India, China, Japan, Egypt, Kenya, Elsinobia, angora, Nanbia, Sudan of Asia, Mexico of America, USA, etc. over 30 tropical, subtropical countries and regions. At present, China has introduced and cultivated India traditional moringa oleifera, India improved moringa oleifera (early-growing and high pod yield) and African moringa oleifera (originally produced only near Kenya Turca lake and southwest Etrussian), and the three varieties are mainly distributed in provinces such as Yunnan, Guizhou, Guangdong, Guangxi, Hainan, Taiwan and the like. At present, the traditional moringa oleifera, the improved moringa oleifera and the african moringa oleifera are mainly eaten. The moringa oleifera can be developed and utilized for the whole body, has wide application, can be used for ornamental, edible and medicinal purposes, can be used for purifying vegetable oil and beer, and is also an ideal oil plant, honey plant, livestock feed and firewood.
The moringa leaves and stems are rich in nutrition, rich in vitamin C which is 6 times that of citrus, carotene which is 4 times that of carrot, calcium and protein which are 4 times and 2 times that of milk respectively, and the contents of potassium, iron, magnesium and the like which are higher than those of other vegetables, fruits and the like. According to the calculation, the moringa oleifera leaf powder with only three tablespoons contains 270% of vitamin A, 42% of protein, 125% of calcium, 70% of iron and 22% of vitamin C which are required by infants every day. The moringa oleifera is a new favorite of the health food industry due to the characteristics of high protein, low fat, high fiber and high vitamin content and special health care effects of reducing blood sugar, resisting bacteria, diminishing inflammation and strengthening heart, and is praised as a new era of health food. The edible moringa oleifera has been recorded hundreds of years ago, has rich nutritional value, is called '21 st century human body dart' in Taiwan province, is recognized as 'national first-push green food' by China green food development center in 2012, and is evaluated as 'special food for state banquet' in the same year. The research results at home and abroad show that the moringa oleifera has rich nutritional value, and the nutritional value is high in seeds, flowers, leaves and stems.
In India and Africa, the moringa oleifera is often used as a traditional medicinal material for treating diseases such as diabetes, hypertension, skin diseases, hypoimmunity, anemia, bone diseases, depression, arthritis, digestive organ tumors and the like, and modern pharmacological researches find that the moringa oleifera has the activities of spasmolysis, antibacterium, antivirus, blood pressure reduction, inflammation diminishing and tumor resisting, and the existence of various active ingredients in the moringa oleifera is very important. The active ingredients mainly refer to fatty acids, flavones, other phenolic components, sugar and glycosides, alkaloids, terpenes and steroids.
However, no research report that the moringa oleifera can improve the intestinal health and promote the intestinal growth through reasonable moringa oleifera nutritional treatment on patients with intestinal failure or short bowel syndrome is found so far.
Disclosure of Invention
The invention aims to provide a moringa oleifera extract with a remarkable effect of promoting intestinal growth.
In order to solve the technical problems, the technical scheme of the invention is the application of the moringa extract in preparing the medicine for promoting the growth of intestinal tract or intestinal villus; the preparation method of the moringa extract comprises the following steps: extracting moringa oleifera with ethanol, extracting the extract with ethyl acetate, and layering to obtain ethyl acetate extract; concentrating the ethyl acetate extract, loading the ethyl acetate extract to a silica gel chromatographic column, eluting by using a petroleum ether-acetone system, then eluting by using methanol, and collecting methanol eluent; concentrating the methanol eluent, loading the concentrated methanol eluent on macroporous adsorption resin, eluting with ethanol, collecting ethanol eluent, concentrating, and drying to obtain the moringa oleifera extract.
Preferably, the extraction is carried out by adopting a soaking or heating reflux extraction mode; the extraction times are 1-5 times, and the extraction time is 1-5 hours each time.
Preferably, the mass percentage concentration of the ethanol is 70-95% during the ethanol extraction.
Preferably, the volume ratio of the petroleum ether to the acetone in the petroleum ether-acetone system is 40-60: 1.
Preferably, the model of the macroporous adsorption resin is D101.
Preferably, the concentration of ethanol in percentage by mass is 70-95% when the ethanol is eluted.
The moringa oleifera extract is obtained by extracting with ethanol and ethyl acetate and then separating with a silica gel chromatographic column and macroporous adsorption resin, and can effectively promote intestinal canal or intestinal villus growth.
Drawings
FIG. 1 is a graph showing the effect of Moringa oleifera extract of the present invention on body length of normal mice;
FIG. 2 is a graph showing the effect of Moringa oleifera extract of the present invention on the intestinal length and body length ratio of normal mice;
FIG. 3 is a graph showing the effect of Moringa oleifera extract of the present invention on the ratio of small intestine length to body length in normal mice;
FIG. 4 is a graph showing the effect of Moringa oleifera extract of the present invention on the length of the large intestine and the body length ratio of normal mice;
FIG. 5 is a photograph showing the effect of low and high dosage groups of moringa extract on the intestinal length of normal mice in a blank control group;
FIG. 6 is a photograph of the effect of a blank control group on the length of normal mouse intestinal villi;
FIG. 7 is a photograph showing the effect of low-dose groups of moringa oleifera extract on the length of intestinal villi in normal mice;
FIG. 8 is a photograph showing the effect of high-dose groups of Moringa oleifera extract on the length of normal mouse intestinal villi;
FIG. 9 shows the effect of low and high dose groups of moringa oleifera extract on the length of intestinal villi in normal mice in a blank control group.
Detailed Description
The following further describes embodiments of the present invention with reference to the drawings. It should be noted that the description of the embodiments is provided to help understanding of the present invention, but the present invention is not limited thereto. In addition, the technical features involved in the embodiments of the present invention described below may be combined with each other as long as they do not conflict with each other.
Example 1
A method for preparing Moringa oleifera extract comprises the following steps: soaking moringa oleifera in 75% ethanol by mass for 2 hours, heating and refluxing the moringa oleifera with 75% ethanol by mass for 2 times, wherein the extraction time is 3 hours each time, combining ethanol soaking solution and ethanol extracting solution, extracting the extracting solution with ethyl acetate, and layering to obtain ethyl acetate extracting solution; concentrating the ethyl acetate extract, loading the ethyl acetate extract to a silica gel chromatographic column, eluting by using a petroleum ether-acetone system with the volume ratio of 50:1, then eluting by using methanol, and collecting methanol eluent; concentrating the methanol eluent, loading the concentrated methanol eluent on D101 macroporous adsorption resin, eluting with 95% ethanol by mass, collecting ethanol eluent, concentrating, and drying to obtain the moringa oleifera extract.
Example 2
A method for preparing Moringa oleifera extract comprises the following steps: soaking moringa oleifera in 95% ethanol for 5 hours, heating and refluxing the moringa oleifera with 795% ethanol for 5 times, wherein the extraction time is 1-5 hours each time, combining the ethanol soaking solution and the ethanol extracting solution, extracting the extracting solution with ethyl acetate, and layering to obtain an ethyl acetate extracting solution; concentrating the ethyl acetate extract, loading the ethyl acetate extract into a silica gel chromatographic column, eluting by using a petroleum ether-acetone system with the volume ratio of 60:1, then eluting by using methanol, and collecting methanol eluent; concentrating the methanol eluent, loading the concentrated methanol eluent on D101 macroporous adsorption resin, eluting with 95% ethanol by mass, collecting ethanol eluent, concentrating, and drying to obtain the moringa oleifera extract.
Example 3
A method for preparing Moringa oleifera extract comprises the following steps: soaking moringa oleifera in 70% ethanol for 1 hour, heating and refluxing the moringa oleifera for 1 time by using the 70% ethanol for extraction for 1 hour, combining ethanol soaking solution and ethanol extracting solution, extracting the extracting solution by using ethyl acetate, and layering to obtain ethyl acetate extracting solution; concentrating the ethyl acetate extract, loading the ethyl acetate extract to a silica gel chromatographic column, eluting by using a 40:1 (volume ratio) petroleum ether-acetone system, then eluting by using methanol, and collecting methanol eluent; concentrating the methanol eluent, loading the concentrated methanol eluent on D101 macroporous adsorption resin, eluting with 70% ethanol by mass percent, collecting ethanol eluent, concentrating and drying to obtain the moringa oleifera extract.
Example 4
A method for preparing Moringa oleifera extract comprises the following steps: soaking moringa oleifera in 90% ethanol for 3 times, each time for 3 hours, to obtain an ethanol extract, extracting the extract with ethyl acetate, and layering to obtain an ethyl acetate extract; concentrating the ethyl acetate extract, loading the ethyl acetate extract to a silica gel chromatographic column, eluting by using a petroleum ether-acetone system with the volume ratio of 50:1, then eluting by using methanol, and collecting methanol eluent; concentrating the methanol eluent, loading the concentrated methanol eluent on D101 macroporous adsorption resin, eluting with 85% ethanol by mass percent, collecting ethanol eluent, concentrating and drying to obtain the moringa oleifera extract.
Example 5
A method for preparing Moringa oleifera extract comprises the following steps: coarsely crushing moringa oleifera, heating and refluxing the crushed moringa oleifera for 3 times by using 75% ethanol in percentage by mass, wherein the extraction time is 3 hours each time to obtain an ethanol extract, extracting the extract by using ethyl acetate, and layering to obtain an ethyl acetate extract; concentrating the ethyl acetate extract, loading the ethyl acetate extract to a silica gel chromatographic column, eluting by using a petroleum ether-acetone system with the volume ratio of 55:1, then eluting by using methanol, and collecting methanol eluent; concentrating the methanol eluent, loading the concentrated methanol eluent on D101 macroporous adsorption resin, eluting with 80% ethanol by mass percentage, collecting ethanol eluent, concentrating, and drying to obtain the moringa oleifera extract.
In order to verify the technical effect of the invention, the following tests are specially carried out:
the experimental animals are KM mice and male mice, and are divided into a blank control group, a moringa oleifera extract high-dose group and a moringa oleifera extract low-dose group, wherein each group comprises 10 mice.
One week after acclimation of the animals, feeding intervention was initiated: the control group was fed with normal feed, the high-dose group was fed with feed prepared by mixing normal feed with moringa extract at a ratio of 1:1, and the low-dose group was fed with feed prepared by mixing normal feed with moringa extract at a ratio of 10: 1.
Animals continuously eat for 65 days, and physiological indexes such as food intake, water intake, weight and the like are detected in the period.
Subjecting mice to CO2Measuring body length after euthanasia, dissecting, taking mouse intestinal tract, measuring total length, small intestine length and large intestine length respectively, injecting precooled pH 7.0 Phosphate Buffered Saline (PBS) into intestinal cavity from one section of intestinal tract by using 20ml injector, cleaning intestinal tract contents, cleaning, and fixing with formalin for 24 h.
As shown in fig. 1 to 9, experimental statistics show that the intestinal tracts of mice fed with the moringa oleifera extract in both the high dose group and the low dose group are significantly increased compared with those in the control group, and the increase ratio is larger after the ratio correction with the body length is performed. The site of growth was found to be primarily in the small intestine, as seen after the smaller and large intestine, respectively. It also can promote intestinal villus growth.
Therefore, the invention proves that the moringa oleifera extract can obviously increase intestinal tracts and mainly increase small intestines through oral ingestion by in vivo animal experiments, and the increase of intestinal villi is found through morphological detection (particularly shown in figures 6-8), so that the extract has good application prospects in the aspects of improving intestinal health and promoting intestinal tract growth.
Wherein, figure 1 is animal body length, figure 2 is the ratio of animal intestinal length to body length, figure 3 is the ratio of animal small intestine length to body length, figure 4 is the ratio of animal large intestine length to body length; FIG. 5 is a photograph of randomly selected animal intestines (the upper intestines of the photograph are blank control group, moringa extract low dose group, and moringa extract high dose group), and the length of the intestines is measured with a 30cm ruler with a precision of 0.1 cm; the photos are taken by randomly selecting one sample in each group and taking a photograph by using a 2000-ten-thousand-pixel lens with a ruler of 5 cm. FIG. 6 is a graph showing the effect of Moringa oleifera extract of the present invention on the length of intestinal villi in normal mice. FIG. 6 is a graph of the effect of a blank control group on the length of normal mouse intestinal villi; FIG. 7 is a graph showing the effect of low-dose groups of moringa oleifera extract on the length of intestinal villi in normal mice; FIG. 8 shows the effect of high-dose groups of Moringa oleifera extract on the length of villi in the small intestine of a normal mouse; fig. 9 is a blank control group, and the influence of the low and high dose groups of the moringa extract on the length of intestinal villi of normal mice (the length of intestinal villi of each group is clearly marked in a chart form, and it can be clearly seen from the chart that the moringa extract obtained by the invention can obviously increase the intestinal villi).
In addition, the ratio of intestinal length to body length is shown in the normal distribution, and the intestinal length of the moringa extract treatment groups with different dosages increases along with the concentration gradient.
The embodiments of the present invention have been described in detail with reference to the accompanying drawings, but the present invention is not limited to the described embodiments. It will be apparent to those skilled in the art that various changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, and the scope of protection is still within the scope of the invention.
Claims (1)
1. Use of Moringa oleifera extract in preparing a medicament for promoting growth of intestinal or intestinal villi; the preparation method of the moringa extract comprises the following steps: extracting moringa oleifera with ethanol, extracting the extract with ethyl acetate, and layering to obtain ethyl acetate extract; concentrating the ethyl acetate extract, loading the ethyl acetate extract to a silica gel chromatographic column, eluting by using a petroleum ether-acetone system, then eluting by using methanol, and collecting methanol eluent; concentrating the methanol eluent, loading the concentrated methanol eluent on macroporous adsorption resin, eluting with ethanol, collecting ethanol eluent, concentrating, and drying to obtain moringa oleifera extract; during the ethanol extraction, the mass percentage concentration of the ethanol is 70-95%;
the extraction is carried out by adopting a soaking or heating reflux extraction mode; the extraction times are 1-5 times, and the extraction time is 1-5 hours each time; the volume ratio of the petroleum ether to the acetone in the petroleum ether-acetone system is 40-60: 1; the model of the macroporous adsorption resin is D101; when the ethanol is eluted, the mass percentage concentration of the ethanol is 70-95%.
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| CN108938690B (en) * | 2018-09-27 | 2021-06-18 | 云南农业大学 | Moringa oleifera leaf ethyl acetate extract and extraction method and application thereof |
| CN111515384B (en) * | 2020-04-10 | 2022-05-24 | 深圳大学 | Nano-silver material mediated and synthesized by moringa seed extract |
| CN116459286B (en) * | 2023-06-07 | 2024-05-14 | 河北瑞龙生物科技有限公司 | Method for extracting moringa flavone from moringa leaves |
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| 辣木叶总黄酮乙醇提取工艺的研究;陈瑞娇;《食品研究与开发》;20070430;第29-31页 * |
| 辣木的化学成分研究;刘长倩等;《安徽农业科学》;20160210(第5期);第142-144页 * |
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