CN114470072A - Ampelopsis grossedentata extract, preparation method thereof and application thereof in preparation of diuretic drugs - Google Patents
Ampelopsis grossedentata extract, preparation method thereof and application thereof in preparation of diuretic drugs Download PDFInfo
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/87—Vitaceae or Ampelidaceae (Vine or Grape family), e.g. wine grapes, muscadine or peppervine
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P7/00—Drugs for disorders of the blood or the extracellular fluid
- A61P7/10—Antioedematous agents; Diuretics
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/12—Antihypertensives
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- 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
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation, decoction
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- 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
- A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- 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
- A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization
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Abstract
The invention provides a vine tea extract, a preparation method thereof and application thereof in preparing diuretic medicines, belonging to the technical field of extraction of natural medicinal components. The ampelopsis grossedentata extract is prepared by extracting ampelopsis grossedentata samples with water and carrying out ultrasonic assistance, wherein the main active ingredients of the ampelopsis grossedentata extract are flavonoids such as dihydromyricetin more than or equal to 280mg/g, myricitrin more than or equal to 10mg/g, myricetin more than or equal to 30mg/g, quercetin more than or equal to 10mg/g, and taxifolin more than or equal to 30 mg/g. The diuretic activity and the nephrotoxicity of the ampelopsis grossedentata extract are evaluated by using an SD rat model, and the result shows that compared with a blank control group, the urine volume of the SD rat in the ampelopsis grossedentata extract gastric perfusion treatment group is remarkably increased by more than or equal to 35% in the period of 2-6 hours, and the SD rat has no nephrotoxicity. The method provided by the invention can realize high-efficiency extraction of the diuretic active ingredients in the vine tea, and further provides a reference for development and utilization.
Description
Technical Field
The invention belongs to the technical field of extraction of natural medicinal components, and particularly relates to a vine tea extract, a preparation method thereof and application thereof in preparation of diuretic medicines.
Background
Hypertension is a leading cause of death and disability in the world. Medical research shows that the intake of sodium is closely related to blood pressure, and the diuretic can excrete sodium to induce diuresis, so that blood volume is reduced, cardiac blood output is reduced, and blood pressure is reduced. At present, the most commonly used antihypertensive drug diuretic has side effects such as metabolic disorder, diuretic resistance and the like, so that the development of a novel diuretic antihypertensive drug without side effects is urgently needed. The natural medicine from the plant plays a role in reducing blood pressure through multiple ways, multiple systems and multiple target points, and has smaller toxic and side effects than western chemical medicines.
The ampelosis grossedentata (hand. -Mazz.) W.T.Wang) which is a new food resource has the effects of clearing away heat and toxic materials, relieving sore throat and swelling, calming the liver and reducing blood pressure, and promoting blood circulation and removing obstruction in channels, and is widely applied to prevention and treatment of diseases such as blood pressure reduction and the like. However, the diuretic antihypertensive chemical active ingredients in the vine tea are not extracted systematically, which hinders the application of the vine tea in developing drugs and functional foods with higher efficacy for lowering blood pressure.
Disclosure of Invention
In view of the above, the present invention aims to provide an ampelopsis grossedentata extract and a preparation method thereof, which can efficiently extract diuretic active ingredients from ampelopsis grossedentata and provide a basis for preparing diuretic drugs.
The invention provides a preparation method of a vine tea extract, which comprises the following steps:
1) mixing vine tea powder with water, and extracting under ultrasound to obtain vine tea crude extract;
2) centrifuging the ampelopsis grossedentata crude extract through a filter bag to obtain a purified ampelopsis grossedentata extract;
3) and freeze-drying the purified vine tea extract to obtain the vine tea extract.
Preferably, the mass ratio of the vine tea powder to the water in the step 1) is 1: (60-70).
Preferably, the ultrasonic treatment time in the step 1) is 40-50 min.
Preferably, the temperature of the ultrasound in the step 1) is 50-55 ℃.
Preferably, the power of the ultrasound in the step 1) is 190W-210W.
Preferably, the particle size of the vine tea powder in the step 1) is less than or equal to 75 microns.
Preferably, when the filter bag is centrifuged in the step 2), the aperture of the filter bag is 1000 meshes, and the rotating speed of the centrifugation is 3000-3500 rpm.
Preferably, the vacuum degree of the freeze drying in the step 3) is 25-30 MPa, the temperature of the freeze drying is-40 to-45 ℃, and the time of the freeze drying is 12-15 h.
The invention provides a vine tea extract prepared by the preparation method, and the active ingredients comprise the following components in percentage by weight: the content of dihydromyricetin is more than or equal to 280mg/g, the content of myricitrin is more than or equal to 10mg/g, the content of myricetin is more than or equal to 30mg/g, the content of quercetin is more than or equal to 10mg/g, and the content of taxifolin is more than or equal to 30 mg/g.
The invention provides application of the vine tea extract in preparing a diuretic medicine.
The preparation method of the vine tea extract provided by the invention comprises the steps of mixing vine tea powder and water, extracting under ultrasonic, centrifuging through a filter bag and freeze-drying to obtain the vine tea extract. The ampelopsis grossedentata extract prepared by the method has high content of active ingredients, and provides a basis for the subsequent preparation of diuretic medicines or foods. Moreover, the preparation method provided by the invention is simple and convenient to operate, has good repeatability, does not adopt toxic and harmful reagents, and is green and environment-friendly.
The invention provides application of the Ampelopsis grossedentata extract prepared by the scheme in preparation of a diuretic medicine or food. The prepared ampelopsis grossedentata extract is applied to rats screened by an Aston diuresis-imitating screening method through gastric lavage, the diuresis effect of the rats is evaluated, the renal toxicity evaluation and other detection results of rats of an ampelopsis grossedentata extract application group show that the diuresis activity is obviously superior to that of other control groups, and the renal function index and the renal HE staining result show that the ampelopsis grossedentata extract has no renal toxicity to SD rats. Therefore, the vine tea extract provided by the invention has excellent diuretic activity and no toxic or side effect, and can be applied to industrial production.
Drawings
FIG. 1 is a structural diagram showing the influence of Ampelopsis grossedentata extract on urine volume of SD rats, which is prepared in example 1 of the present invention;
FIG. 2 shows the effect of Ampelopsis grossedentata extract on the level of renal function index in SD rats and HE stained tissue sections in example 1 of the present invention, wherein 1, Ampelopsis grossedentata extract low dose group, normal diet + Ampelopsis grossedentata extract (0.5 g/kg); 2. vine tea extract high dose group, normal feed + vine tea extract (2.5 g/kg); 3. blank experiment group, normal feed + normal saline; A. d, G is blank group; B. e, H is low dose intragastric group containing Ampelopsis grossedentata extract; C. f, K is tissue of kidney cortex region of Ampelopsis grossedentata extract in high dose intragastric group, with magnification of 10 ×,20 ×, and 40 ×).
Detailed Description
The invention provides a preparation method of a vine tea extract, which comprises the following steps:
1) mixing vine tea powder with water, and extracting under ultrasound to obtain vine tea crude extract;
2) centrifuging the ampelopsis grossedentata crude extract through a filter bag to obtain a purified ampelopsis grossedentata extract;
3) and freeze-drying the purified vine tea extract to obtain the vine tea extract.
The method mixes vine tea powder with water, and extracts the vine tea powder under ultrasound to obtain vine tea crude extract.
In the present invention, the particle size of the ampelopsis grossedentata powder is an important factor that affects the extraction of active ingredients. Too large particle size and insufficient solvent action; the granularity is too small, the cost is consumed, and the processing personnel are not good. Experiments prove that the grain size of the vine tea powder is preferably less than or equal to 75 mu m, and the vine tea powder has great help for improving the extraction of diuretic active ingredients in the extract.
In the invention, the mass ratio of the vine tea powder to water is preferably 1: (60-70), more preferably 1: 65. The present invention is not particularly limited in kind of water, and water known in the art, such as purified water, distilled water, etc., may be used. The ultrasonic treatment time is preferably 40-50 min, and more preferably 45 min. The ultrasonic time has an influence on the extraction effect of the active ingredients in the extract, and the extraction of the diuretic active ingredients is not facilitated by too high or too low ultrasonic time. The temperature of the ultrasound is preferably 50-55 ℃, and more preferably 52 ℃. The ultrasonic temperature has an influence on the extraction effect of the active ingredients in the extract, and too high or too low temperature is not beneficial to the extraction of the diuretic active ingredients. The power of the ultrasound is preferably 190W to 210W, more preferably 200W. The power of the ultrasound also has an effect on the extraction effect of the active ingredients in the extract, and too high or too low ultrasound power is not beneficial to the extraction of the diuretic active ingredients.
After the vine tea crude extract is obtained, the vine tea crude extract is centrifuged by a filter bag to obtain a purified vine tea extract.
In the present invention, when the filter bag is centrifuged, the pore size of the filter bag is preferably 1000 mesh. The rotation speed of centrifugation is preferably 3000-3500 rpm, and more preferably 3300 rpm. The purpose of the bag centrifugation is to remove solid foreign particles.
After the purified vine tea extract is obtained, the purified vine tea extract is subjected to freeze drying to obtain the vine tea extract.
In the present invention, the vacuum of the freeze-drying is preferably 25 to 30MPa, and more preferably 28 MPa. The temperature of the freeze drying is preferably-40 to-45 ℃, and more preferably-42 ℃. The freeze drying time is preferably 12-15 h, and more preferably 13 h.
The invention provides a vine tea extract prepared by the preparation method, and the active ingredients comprise the following components in percentage by weight: the content of dihydromyricetin is more than or equal to 280mg/g, the content of myricitrin is more than or equal to 10mg/g, the content of myricetin is more than or equal to 30mg/g, the content of quercetin is more than or equal to 10mg/g, and the content of taxifolin is more than or equal to 30 mg/g.
The invention provides application of the vine tea extract in preparing a diuretic medicine.
In the invention, renal function indexes of urea, creatinine and urea/creatinine are evaluation standards, and the diuretic activity of the vine tea extract prepared by the invention is also obviously superior to that of the vine tea extract prepared by a comparative example. Meanwhile, experiments prove that the ampelopsis grossedentata extract has no nephrotoxicity to SD rats and can be used for industrial production.
In the present invention, the dosage form of the drug is preferably an oral preparation. The present invention is not particularly limited in kind of the oral preparation, and oral preparations known in the art, for example, oral liquid, tablets, pills, etc. may be used. The content of the vine tea extract in the medicine is preferably 50-60% by mass, and more preferably 55% by mass. The medicine also comprises a certain amount of pharmaceutically acceptable auxiliary materials. The type of the auxiliary materials is adjusted according to the dosage form. The method for preparing the medicine is not particularly limited, and the method for preparing the medicine known in the field can be adopted. 1-2 g of the medicine is poured into a cup and dissolved in 100-200 mL of boiled water for drinking.
The ampelopsis grossedentata extract, the preparation method thereof and the application thereof in preparing diuretic drugs provided by the present invention will be described in detail with reference to the following examples, but they should not be construed as limiting the scope of the present invention.
Experimental raw materials, consumables and instrument source description:
the ampelopsis grossedentata raw material: biche Biba tea professional cooperative society of Xifeng county;
purified water: nyshi yunwishan drinks, ltd;
a pulverizer: guangdong Zhonghuaheng science and technology, Inc.;
separating and screening: tengde wire mesh products, Inc., Anping county;
an ultrasonic instrument: kunshanshumei ultrasonic instruments ltd;
a freeze dryer: ningbo Xinzhi Biotechnology GmbH;
a centrifuge: zhanggang city happy surplus motor equipment factory;
a filter bag: fine Kokok textile printing and dyeing Equipment Co., Ltd, Fushan City.
Example 1
(1) In 23 days 5 months in 2020, 10kg of vine tea raw material is purchased from Biche coerulescens of Xifeng county, the vine tea raw material is crushed by a crusher (ZGP-2320Q) and is sieved by a 200-mesh sieve, and the crushed particle size of the vine tea sample is less than or equal to 75 mu m;
(2) the method is characterized by using a kq5200e ultrasonic instrument, using purified water as an extraction solvent, and mixing the raw materials according to a material-liquid ratio of 1: 60 (the proportion of the vine tea to the water solution), and carrying out ultrasonic treatment for 40min at 50 ℃ and with the power of 190w to obtain an extracting solution;
(3) centrifuging the extractive solution with a filter bag of 3000r/min and 1000 meshes by use of a centrifuge (model 200);
(4) lyophilizer (SCIENTZ-30F), conditions of lyophilization: vacuum degree of 25MPa, temperature of-40 deg.C, and time of 12 hr, lyophilizing Ampelopsis Grossdentata extract to obtain Ampelopsis Grossdentata extract with diuretic activity.
(5) The content of main flavonoid active ingredients including dihydromyricetin, myricitrin, myricetin, quercetin and taxifolin in the vine tea extract is detected according to a detection method provided by a patent of detection methods of the types and content of flavonoid compounds in vine tea (patent number: ZL 201910739376.7).
Example 2
(1) In 11 days 6 and 2020, 9.8kg of vine tea raw material is purchased from Biche camara specialty cooperative, Xifeng county, and is crushed by a crusher (ZGP-2320Q), and is sieved by a 400-mesh sieve, so that the crushed particle size of the vine tea sample is less than or equal to 38 mu m;
(2) the method is characterized by using a kq5200e ultrasonic instrument, using purified water as an extraction solvent, and mixing the raw materials according to a material-liquid ratio of 1: 60 (the proportion of the vine tea to the water solution), and carrying out ultrasonic treatment for 50min at the temperature of 55 ℃ and the power of 210w to obtain an extracting solution;
(3) centrifuging the extractive solution with a filter bag of 3000r/min and 1000 meshes by use of a centrifuge (model 200);
(4) lyophilizer (SCIENTZ-30F), conditions of lyophilization: vacuum degree of 25MPa, temperature of-45 deg.C, and time of 12 hr, lyophilizing Ampelopsis Grossdentata extract to obtain Ampelopsis Grossdentata extract with diuretic activity.
(5) The content of main flavonoid active ingredients including dihydromyricetin, myricitrin, myricetin, quercetin and taxifolin in the vine tea extract is detected according to a detection method provided by a patent of detection methods of the types and content of flavonoid compounds in vine tea (patent number: ZL 201910739376.7).
Example 3
(1) 7 months and 7 days in 2020, 10.4kg of vine tea raw material is purchased from Biche camara specialty cooperative of Xifeng county, ZGP-2320Q, crushed and sieved by a 200-mesh sieve, and the crushed particle size of the vine tea sample is less than or equal to 75 mu m;
(2) the method is characterized by using a kq5200e ultrasonic instrument, using purified water as an extraction solvent, and mixing the raw materials according to a material-liquid ratio of 1: 70 (the ratio of the vine tea to the water solution), and carrying out ultrasonic treatment for 45min at the temperature of 55 ℃ and the power of 200w to obtain an extracting solution;
(3) centrifuging the extractive solution with a centrifuge (model 200) and a filter bag of 3300r/min and 1000 meshes;
(4) lyophilizer (SCIENTZ-30F), conditions of lyophilization: vacuum degree of 30MPa, temperature of-48 deg.C, and time of 15 hr, lyophilizing Ampelopsis Grossdentata extract to obtain Ampelopsis Grossdentata extract with diuretic activity.
(5) The content of main flavonoid active ingredients including dihydromyricetin, myricitrin, myricetin, quercetin and taxifolin in the vine tea extract is detected according to a detection method provided by a patent of detection methods of the types and content of flavonoid compounds in vine tea (patent number: ZL 201910739376.7).
Example 4
(1) In 16 days 8 months in 2020, 11.1kg of vine tea raw material is purchased from Biche camara specialty cooperative of Xifeng county, ZGP-2320Q, crushed and sieved by a 200-mesh sieve, and the crushed particle size of the vine tea sample is less than or equal to 75 μm;
(2) the method is characterized by using a kq5200e ultrasonic instrument, using purified water as an extraction solvent, and mixing the raw materials according to a material-liquid ratio of 1: 70 (the ratio of the vine tea to the water solution), and carrying out ultrasonic treatment for 45min at the temperature of 55 ℃ and the power of 200w to obtain an extracting solution;
(3) centrifuging the extractive solution with 3500r/min 1000 mesh filter bag by using centrifuge (model 200);
(4) lyophilizer (SCIENTZ-30F), conditions of lyophilization: vacuum degree of 28MPa, temperature of-45 deg.C, and time of 15 hr, lyophilizing Ampelopsis Grossdentata extract to obtain Ampelopsis Grossdentata extract with diuretic activity.
(5) The content of main flavonoid active ingredients including dihydromyricetin, myricitrin, myricetin, quercetin and taxifolin in the vine tea extract is detected according to a detection method provided by a patent of detection methods of the types and content of flavonoid compounds in vine tea (patent number: ZL 201910739376.7).
Comparative example 1
Compared with the example 1, only the step (1) is changed, and the vine tea sample is sieved by a 100-mesh sieve, so that the crushed particle size of the vine tea sample is less than or equal to 150 mu m.
Comparative example 2
Compared with example 1, only step (2) was changed, and the sonication time was 30 min.
Comparative example 3
Compared with the example 1, only the step (2) is changed, and the ultrasonic time is 60 min.
Comparative example 4
Compared with example 1, only step (2) was changed, the temperature was 60 ℃.
Comparative example 5
Compared with the embodiment 1, only the step (2) and the power 180w are changed.
Comparative example 6
Compared with the embodiment 1, only the step (2) and the power 220w are changed.
Comparative example 7
Compared with the example 1, only the step (3) is changed, and the centrifugal rotating speed is set to be 2500 r/min.
Comparative example 8
Compared with the embodiment 1, only the step (3) is changed, and the centrifugal rotating speed is set to be 4000 r/min.
Comparative example 9
Only step (4) was changed to set a degree of vacuum of 20MPa as compared with example 1.
Comparative example 10
Only step (4) was changed to set a vacuum degree of 35MPa as compared with example 1.
Comparative example 11
Only step (4) was changed as compared with example 1, and the temperature was set at-60 ℃.
Comparative example 12
Compared with example 1, only step (4) was changed, and the setting time was 10 h.
Comparative example 13
Compared with example 1, only step (4) was changed, and the setting time was 17 h.
Example 5
For the active ingredient extract obtained in the test, the extract yield is calculated by adopting the following formula I:
the content of main flavone active ingredients is detected by adopting a detection method provided by a patent of detection method of flavonoid compound types and content in vine tea (patent number: ZL201910739376.7), and a specific formula is shown in a formula II.
The vine tea extract yield and the content of main flavonoid active ingredients of examples 1, 2, 3 and 4 have no significant difference. Compared with the comparative example, the method provided by the invention can obviously improve the yield of the active extract in the example and obviously improve the content of the main flavonoid active ingredient.
TABLE 1 Ampelopsis grossedentata extract yield and major flavonoid active ingredient content
Note: different lower case letters in the same column represent significant differences in data.
Example 6
Animal experiments were carried out on Ampelopsis grossedentata extracts obtained in examples 1 to 4 and comparative examples 1 to 13, and rats were screened before administration by an imitation Aston diuresis screening method (literature source: Chenjing, Sun Yun Chao, ran Xiaokuo, Yuan, sinus Qiang. research on diuresis of Atractylodes macrocephala [ J ]. modern Chinese medicine 2016,18(05):563-567.) and healthy male SD rats weighing 180 to 200g were selected for evaluation of renal toxicity function. The breeding environment is a barrier environment, the temperature is 26 + -2 deg.C, the relative humidity is 55%, and the day and night cycle time is 12 h. Animals are raised in cages, each cage is provided with 8 animals, and the animals can freely drink and eat water. Respectively intragastrically infusing Ampelopsis grossedentata extracts with different dosages: low dose (10mg/kg), high dose (50 mg/kg); the control group was gavaged with equal volume of normal saline. Immediately after administration, rats were placed in metabolic cages and urine volume was measured 1 time every 1h for a total of 6 h. Evaluation of renal toxicity: the blood is taken from the eyeball to detect the renal function index once every 4 treatment courses (30 days): urea (blood urea), creatinine (creatinine), urea/creatinine (blood urea/creatinine). The uric acid detection is carried out 3 times according to the reference standard WS/T53-1996 and the creatinine detection is carried out according to the reference standard WS/T98-1996. Dissecting and picking the kidney, embedding paraffin, HE staining, slicing, and observing the state of the tissue by panoramic scanning. Three groups of indexes related to kidney function before and after administration are analyzed in a comparative way, and the kidney toxicity of the vine tea extract (less than or equal to 2.5g/kg) to SD rats is evaluated.
TABLE 2 Ampelopsis grossedentata extract yield and effect on renal function index in Normal SD rats
Note: different lower case letters in the same column represent significant differences in data.
The main renal function indexes of examples 1, 2, 3 and 4 are urea, creatinine and urea/creatinine without significant difference. The method of the present invention can make the diuretic activity of the examples significantly better than that of the other examples as well, compared with the comparative examples. Meanwhile, the renal function indexes of urea, creatinine and urea/creatinine in the embodiment have no significant difference between the change of the renal function indexes before and after administration. In the renal HE staining, the renal tissues (glomeruli and renal tubules) of the examples are all structurally intact, and the cells are clearly stained. The cortex and medulla of the kidney tissue are obviously demarcated, more glomeruli are concentrated in the cortex area, and fewer glomeruli are in the medullary area. The boundaries of the glomeruli are evident and capillary vessels within the glomeruli are observed. The renal tubule lumen has clear boundary, no inflammatory cell infiltration and mesenteric stromal cell proliferation, no atrophy or expansion, obvious difference between the proximal convoluted tubule and the distal convoluted tubule, star-shaped radial proximal convoluted tubule lumen, and elliptic or circular distal convoluted tubule lumen (figure 2). The process for obtaining the ampelopsis grossedentata extract with different dosages has no nephrotoxicity to SD rats. The method provided by the invention is simple and convenient to operate, low in cost, good in repeatability and applicable to production.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (10)
1. A preparation method of a vine tea extract is characterized by comprising the following steps:
1) mixing vine tea powder with water, and extracting under ultrasound to obtain vine tea crude extract;
2) centrifuging the ampelopsis grossedentata crude extract through a filter bag to obtain a purified ampelopsis grossedentata extract;
3) and freeze-drying the purified vine tea extract to obtain the vine tea extract.
2. The preparation method according to claim 1, wherein the mass ratio of the vine tea powder to the water in the step 1) is 1: (60-70).
3. The preparation method according to claim 1, wherein the time of the ultrasonic treatment in the step 1) is 40 to 50 min.
4. The method according to claim 1, wherein the temperature of the ultrasound in step 1) is 50 to 55 ℃.
5. The method for preparing the ultrasonic probe according to claim 1, wherein the power of the ultrasonic in the step 1) is 190W to 210W.
6. The preparation method according to claim 1, wherein the particle size of the vine tea powder in step 1) is less than or equal to 75 μm.
7. The preparation method of claim 1, wherein when the filter bag is centrifuged in the step 2), the aperture of the filter bag is 1000 meshes, and the rotation speed of the centrifugation is 3000-3500 rpm.
8. The preparation method according to any one of claims 1 to 7, wherein the degree of vacuum of the freeze drying in the step 3) is 25 to 30MPa, the temperature of the freeze drying is-40 to-45 ℃, and the time of the freeze drying is 12 to 15 hours.
9. The Ampelopsis grossedentata extract prepared by the preparation method according to any one of claims 1 to 8, wherein the active ingredients comprise the following components in percentage by weight: the content of dihydromyricetin is more than or equal to 280mg/g, the content of myricitrin is more than or equal to 10mg/g, the content of myricetin is more than or equal to 30mg/g, the content of quercetin is more than or equal to 10mg/g, and the content of taxifolin is more than or equal to 30 mg/g.
10. Use of the Ampelopsis grossedentata extract of claim 9 for the preparation of a diuretic.
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