阅读说明：本技术 一种抗糖尿病肾病的女贞子提取物及制备方法和应用 (Glossy privet fruit extract for resisting diabetic nephropathy and preparation method and application thereof ) 是由 张学兰 栾茹乔 李慧芬 吴鹏 于 2020-06-02 设计创作，主要内容包括：本发明公开了一种抗糖尿病肾病的女贞子提取物,通过向女贞子中加入黄酒和/或水,隔水蒸制10～30h,取出女贞子,50～80℃干燥3～8h,粉碎成粗粉,加入粗粉30～80wt%的乙醇超声提取2～3次,每次0.5～2h,合并滤液,减压回收乙醇至无醇味,得浸膏,浸膏加水分散,上AB-8型大孔树脂,依次用5BV水洗脱和5BV体积浓度50%的乙醇溶液以2BV/h流速洗脱,收集乙醇洗脱液,回收乙醇,冷冻干燥得女贞子提取物。本发明女贞子活性成分转化后,提升了女贞子的利用价值,活性强,可开发成为治疗DN药物或保健食品,制成各种剂型,方便临床应用。且提取的复合成分效果要好于红景天苷,避免了传统提取红景天苷提取程序复杂,解决了红景天苷治疗效果有限的问题。(The invention discloses a glossy privet fruit extract for resisting diabetes and nephropathy, which is prepared by adding yellow wine and/or water into glossy privet fruit, steaming for 10-30 hours in a water-proof manner, taking out the glossy privet fruit, drying for 3-8 hours at 50-80 ℃, crushing into coarse powder, adding 30-80 wt% of ethanol into the coarse powder, performing ultrasonic extraction for 2-3 times, each time for 0.5-2 hours, combining filtrates, recovering ethanol under reduced pressure until no alcohol smell exists, obtaining extract, adding water into the extract for dispersing, adding AB-8 type macroporous resin, sequentially eluting with 5BV water and 5BV ethanol solution with volume concentration of 50% at the flow rate of 2BV/h, collecting ethanol eluent, recovering ethanol, and performing freeze drying to obtain the glossy privet fruit extract. After the active ingredients of the glossy privet fruit are converted, the utilization value of the glossy privet fruit is improved, the glossy privet fruit has strong activity, and the glossy privet fruit can be developed into medicaments for treating DN or health-care food to be prepared into various formulations, thereby being convenient for clinical application. The effect of the extracted composite components is better than that of salidroside, so that the complex extraction procedure of the traditional extraction of salidroside is avoided, and the problem of limited curative effect of salidroside is solved.)

1. A glossy privet fruit extract for resisting diabetic nephropathy is characterized by being prepared by the following method:

(1) adding yellow wine and/or water into glossy privet fruit, steaming for 10-30 h in a water-proof manner, taking out the glossy privet fruit, drying for 3-8 h at 50-80 ℃, and crushing into coarse powder;

(2) adding ethanol with the amount of 30-80 wt% of the coarse powder into the coarse powder in the step (1), performing ultrasonic extraction for 2-3 times, each time for 0.5-2 hours, filtering, combining filtrates, and recovering ethanol under reduced pressure until no ethanol smell exists, thereby obtaining an extract;

(3) dispersing the extract with water, loading onto AB-8 type macroporous resin, sequentially eluting with 5BV water and 5BV 50% ethanol solution at flow rate of 2BV/h, collecting ethanol eluate, recovering ethanol under reduced pressure, and freeze drying to obtain fructus Ligustri Lucidi extract.

2. The fructus ligustri lucidi extract of claim 1, wherein the fructus ligustri lucidi in step (1) is dry fructus ligustri lucidi decoction pieces, 10-30 g yellow wine and 50-100 g water are added to each 100g fructus ligustri lucidi decoction pieces.

3. The fructus Ligustri Lucidi extract as claimed in claim 1, wherein the fructus Ligustri Lucidi in step (1) is placed in a stainless steel container and steamed in water, and the steaming temperature is 80-100 ℃.

4. The fructus ligustri lucidi extract as claimed in claim 1, wherein the drying in step (1) is performed by a hot air circulation drying oven.

5. The fructus Ligustri Lucidi extract as claimed in claim 1, wherein the ethanol extract in step (2) is 50% ethanol solution.

6. A method for preparing the extract of fructus Ligustri Lucidi for treating diabetic nephropathy according to any one of claims 1 to 5, comprising the steps of:

(1) adding yellow wine and/or water into glossy privet fruit, steaming for 10-30 h in a water-proof manner, taking out the glossy privet fruit, drying for 3-8 h at 50-80 ℃, and crushing into coarse powder;

(2) adding ethanol with the amount of 30-80% of the coarse powder into the coarse powder in the step (1), performing ultrasonic extraction for 2-3 times, each time for 0.5-2 hours, filtering, combining filtrates, and recovering ethanol under reduced pressure until no ethanol smell exists, thereby obtaining an extract;

(3) dispersing the extract with water, loading onto AB-8 type macroporous resin, sequentially eluting with 5BV water and 5BV 50% ethanol solution at flow rate of 2BV/h, collecting ethanol eluate, recovering ethanol under reduced pressure, and freeze drying to obtain fructus Ligustri Lucidi extract.

7. The application of the glossy privet fruit extract for resisting diabetic nephropathy in any one of claims 1-5, which is used for preparing a medicament or health product for resisting diabetic nephropathy.

8. The use of claim 7, wherein the anti-diabetic nephropathy pharmaceutical or health product is in the form of tablets, capsules, granules, pills, pellets, powder, dripping pills, decoction, syrup, mixture, soft extract or extract.

Technical Field

The invention belongs to the field of extraction of effective components of traditional Chinese medicines, and particularly relates to a glossy privet fruit extract for resisting diabetic nephropathy, and a preparation method and application thereof.

Background

Diabetic Nephropathy (DN) is a common diabetic microvascular complication and one of the leading causes of death in diabetic patients. In recent years, the incidence rate of DN is on an increasing trend, but no effective medicine for treating DN exists at present. The traditional Chinese medicine contains various effective components, can generate comprehensive treatment effect through multiple ways and multiple targets, and has obvious advantages and characteristics for preventing and treating chronic metabolic diseases. The intensive research on the traditional Chinese medicine with exact efficacy for treating DN is helpful for better developing new medicaments for treating DN.

Fructus Ligustri Lucidi is Ligustrum lucidum belonging to OleaceaeLigustrum lucidumThe dry mature fruit of ait has sweet and bitter taste and cool property, has the functions of nourishing liver and kidney, improving eyesight and blackening hair, and is a traditional Chinese medicine for nourishing liver and kidney which is commonly used in clinic. In traditional Chinese medicine, the medicine is used for treating internal heat and diabetes, is commonly used for diabetes and complications thereof, and is a core medicine for treating DN. The processing method for women in the past mainly comprises steaming, wine mixing, nine times of steaming and nine times of drying in the sun and the like, and the women are processed mainly by wine steaming and wine stewing at present. The prescription of "Chinese pharmacopoeia" 2015 is recorded with two processing specifications of glossy privet fruit and wine glossy privet fruit. Long-term clinical practice proves that the glossy privet fruit can relieve the cold and slippery property of the glossy privet fruit after being processed with wine and enhance the effect of nourishing liver and kidney, so the glossy privet fruit is mainly taken as a wine product for medicine in clinical all the times. Fructus Ligustri Lucidi mainly contains iridoid glycosides, triterpenes, and phenethyl alcohol as effective components. Iridoid glycoside compounds separated from fructus Ligustri Lucidi mainly include specnuezhenide (specnuezhenide), nuezhenide G13 (G13), oleonuezhenide, nuezhendic acid (nuezhenic acid), neonuezhenide (neujuzhenide), etc., triterpenes mainly include oleanolic acid (oleanolic acid), ursolic acid (ursolic acid), etc., phenethyl alcohol mainly includes salidroside (salidroside), hydroxytyrosol (hydroxytyrosol), tyrosol (tyrosol), verbascoside (acteoside), echinacoside (echinacoside), etc. The glossy privet is also a common greening tree and a common street tree, and a large amount of mature glossy privet falls on the ground in autumn every year to dye the ground black, thereby causing environmental pollution and pollutionThe resources are wasted.

Salidroside is one of main active ingredients of rhodiola rosea of Crassulaceae. According to the report of the literature, salidroside has antioxidant function, can reduce ROS generated by cells, remove free radicals, and improve the activity of antioxidant enzymes such as CAT and SOD. The research proves that salidroside can inhibit proliferation of high-sugar-induced mesangial cells, and can also play a role in protecting the ischemia-reperfusion injury of the kidney and chronic kidney diseases. Research has shown that rhodiola rosea has the function of treating or inhibiting DN. Rhodiola plants grow in plateau areas, are poor in resources and expensive in price, and as a substitute thereof, traditional Chinese medicine glossy privet fruit contains various active ingredients, wherein salidroside is contained, but the following problems also exist: the content of salidroside in natural fructus Ligustri Lucidi is limited, and the active ingredients in fructus Ligustri Lucidi are various, and the steps for separating salidroside are complicated.

At present, no report on an extract with DN resistance obtained by extracting and separating glossy privet fruit serving as a raw material is found. In order to fully utilize the resources of glossy privet fruits and reduce the resource waste and the environmental pollution, it is necessary to carry out deep research on the active ingredients and the pharmacological action of the glossy privet fruits, so as to expand the medicinal approach of the glossy privet fruits and provide guidance for the clinical application of the glossy privet fruits.

Disclosure of Invention

Aiming at the problems of resource waste of glossy privet fruits and high price of rhodiola rosea in the prior art, the invention provides a DN (DN) -resistant glossy privet fruit extract and a preparation method and application thereof.

The invention is realized by the following technical scheme:

a glossy privet fruit extract for resisting diabetic nephropathy is prepared by the following method:

(1) adding yellow wine and/or water into glossy privet fruit, steaming for 10-30 h in a water-proof manner, taking out the glossy privet fruit, drying for 3-8 h at 50-80 ℃, and crushing into coarse powder;

(2) adding ethanol with the amount of 30-80 wt% of the coarse powder into the coarse powder in the step (1), performing ultrasonic extraction for 2-3 times, each time for 0.5-2 hours, filtering, combining filtrates, and recovering ethanol under reduced pressure until no ethanol smell exists, thereby obtaining an extract;

(3) dispersing the extract with water, loading onto AB-8 type macroporous resin, sequentially eluting with 5BV water and 5BV 50% ethanol solution at flow rate of 2BV/h, collecting ethanol eluate, recovering ethanol under reduced pressure, and lyophilizing to obtain fructus Ligustri Lucidi extract.

Further, the glossy privet fruit in the step (1) is dry glossy privet fruit decoction pieces, and 10-30 g of yellow wine and 50-100 g of water are added into each 100g of glossy privet fruit decoction pieces.

Further, the glossy privet fruit in the step (1) is put into a stainless steel container to be steamed in a water-proof way, and the steaming temperature is 80-100 ℃.

Further, the drying in the step (1) adopts a hot air circulation drying oven for drying.

Further, the ethanol extract in the step (2) is a 50% ethanol solution.

In the invention, the preparation method of the glossy privet fruit extract for resisting diabetic nephropathy comprises the following steps:

(1) adding yellow wine and/or water into glossy privet fruit, steaming for 10-30 h in a water-proof manner, taking out the glossy privet fruit, drying for 3-8 h at 50-80 ℃, and crushing into coarse powder;

(2) adding ethanol with the amount of 30-80% of the coarse powder into the coarse powder in the step (1), performing ultrasonic extraction for 2-3 times, each time for 0.5-2 hours, filtering, combining filtrates, and recovering ethanol under reduced pressure until no ethanol smell exists, thereby obtaining an extract;

(3) dispersing the extract with water, loading onto AB-8 type macroporous resin, sequentially eluting with 5BV water and 5BV 50% ethanol solution at flow rate of 2BV/h, collecting ethanol eluate, recovering ethanol under reduced pressure, and freeze drying to obtain fructus Ligustri Lucidi extract.

In the invention, the application of the glossy privet fruit extract for resisting diabetic nephropathy is used for preparing a medicament or a health-care product for resisting diabetic nephropathy; the invention fully utilizes the glossy privet fruit medicinal material resources, and researches the active ingredients of the glossy privet fruit to prepare the glossy privet fruit extract with high active ingredient content and strong DN resisting activity, has a protective effect on DN rats, has little adverse reaction, and is expected to be developed into new DN resisting medicines and health care products.

Furthermore, the anti-diabetic nephropathy medicine or health product is in the dosage forms of tablets, capsules, granules, pills, pellets, powder, dripping pills, decoction, syrup, mixture, soft extract or extract.

Advantageous effects

(1) The invention mainly takes the glossy privet fruit as the raw material, after the active ingredients are converted, the active site of the glossy privet fruit is extracted, the utilization value of the glossy privet fruit is improved, the comprehensive utilization of glossy privet fruit resources is realized, the refined active site of the glossy privet fruit for resisting DN is developed, the activity is strong, the glossy privet fruit can be developed into a new medicament or health food for treating DN, and the medicaments in various dosage forms can be prepared by convenient and pharmaceutically acceptable carriers, thereby facilitating the clinical application.

(2) The chemical components and pharmacological activity of the glossy privet fruit are researched and screened, so that the content of the active components of the glossy privet fruit extract obtained by the invention is up to more than 50%; can remarkably reduce the blood sugar, urine protein, Serum Creatinine (SCR) and urea nitrogen (BUN) of DN rats, can adjust the blood lipid disorder of the DN rats, improve the atrophy and the sclerosis of glomeruli, and relieve the adiposis and inflammatory cell infiltration of renal tubular epithelial cells; and the effect of the extracted compound components is better than that of the single salidroside, and the synergistic effect of the compound components is further proved.

(3) On the basis of fully utilizing the traditional Chinese medicine resources of the glossy privet fruit, the method not only avoids the problem of complex extraction procedure of the traditional extraction of the salidroside, but also solves the problem of limited curative effect of the salidroside, and the extracted compound active ingredient has better curative effect on DN.

Drawings

FIG. 1 is a high performance liquid chromatogram of the extract of Ligustrum lucidum in example 1;

FIG. 2 is a high performance liquid chromatogram of the extract of Ligustrum lucidum of comparative example 1, hydroxytyrosol 2, Salidroside 3, tyrosol 4, Echinacoside 5, Ligustrum neoleucum glycoside 6, verbascoside 7, Ligustrum teucrium glycoside 8, Ligustric acid 9, Ligustrum lucidum glycoside G1310, oleoneuzhenide;

FIG. 3 is a kidney histomorphogram (X200) for each group of rats.

Detailed Description

For a further understanding of the invention, reference will now be made to the preferred embodiments of the invention in conjunction with the following examples, but it will be understood that the description is intended to illustrate the features and advantages of the invention further, and not to limit the invention.