CN110787240A - Application of longan shell extract in preparing composition for regulating and controlling SREBP-1c, ACC and SCD-1 proteins - Google Patents
Application of longan shell extract in preparing composition for regulating and controlling SREBP-1c, ACC and SCD-1 proteins Download PDFInfo
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- CN110787240A CN110787240A CN201810860922.8A CN201810860922A CN110787240A CN 110787240 A CN110787240 A CN 110787240A CN 201810860922 A CN201810860922 A CN 201810860922A CN 110787240 A CN110787240 A CN 110787240A
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- longan
- longan shell
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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)
- A61K36/77—Sapindaceae (Soapberry family), e.g. lychee or soapberry
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- 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
- A23L33/17—Amino acids, peptides or proteins
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
Abstract
The invention relates to application of a longan shell extract, in particular to application of the longan shell extract in preparing a composition for regulating and controlling SREBP-1c, ACC and SCD-1 protein. The invention provides application of a longan shell extract in preparing a composition for reducing expression of SREBP-1c, ACC and SCD-1 protein, wherein the longan shell extract is obtained by extracting longan shells with water, alcohol or a solvent containing water and alcohol. The longan shell extract can obviously inhibit lipogenesis, reduce GPT index and protect liver.
Description
Technical Field
The invention relates to application of a longan shell extract, in particular to application of the longan shell extract in preparing a composition for regulating and controlling SREBP-1c, ACC and SCD-1 protein.
Background
The liver diseases of Taiwan population are caused by virus infection, drug abuse, long-term alcoholism, etc. According to the statistics data of the health administration of Taiwan administrative department in 2015, the liver diseases such as chronic liver diseases and liver cirrhosis are shown to be in the tenth position of ten major causes of death in Taiwan.
In recent years, the consumption of alcoholic beverages in taiwan has been greatly increased, and the incidence of alcoholic liver diseases has also been relatively increased. After long-term alcohol intake, the biochemical and pathological injuries induced in the human body are very complex, and among them, the tissue most deeply affected is the liver, the main tissue of alcohol metabolism. Liver damage from long-term alcohol consumption includes: fatty liver, alcoholic hepatitis, cirrhosis and the like, and liver cancer is caused if the treatment is not performed in time.
Fatty liver is the earliest sign of alcoholic liver disease. Studies have shown that patients with alcoholic fatty liver have a greatly increased probability of developing liver fibrosis and cirrhosis, and studies have indicated that fatty liver is associated with liver cancer. Therefore, if fatty liver can be improved, the chances of hepatic fibrosis and cirrhosis can be reduced. However, there is no effective treatment method for fatty liver, and most of the clinical suggestions are that patients eat less food with high fat content, take more vegetables and fruits, exercise properly and keep normal work, and most importantly, the patients are prohibited from taking alcohol. However, it is very difficult to withdraw alcohol in a short time for patients suffering from alcohol dependence of fatty liver, and the maximum therapeutic effect can be achieved if the planned reduction of alcohol intake is accompanied by consumption of food capable of reducing fatty liver. Since the liver is an important organ of the human body responsible for detoxification or metabolism. Alcohol abuse and hepatitis virus infection are two main causes of liver diseases, so that the search for antioxidant foods for preventing or alleviating liver damage is an important issue in nutrition medicine.
The longan is produced in 7-9 months, the production place is in Tainan, and the annual output of the whole province is about one hundred thousand to thirteen thousand tons. The longan is baked and peeled to obtain the longan which can be used as a dietary supplement medicinal material and can be stored for a long time. However, after processing longan, a large amount of longan shell byproducts are generated. If these by-products, which can only be discarded, can be further effectively utilized and have a health-care effect on liver diseases, they are a great welfare for human beings.
Disclosure of Invention
In order to solve the above problems, the present invention provides a use of a longan shell extract for preparing a composition for regulating SREBP-1c and ACC and SCD-1 protein, wherein the longan shell extract is obtained by extracting longan shell with water, alcohol, or a solvent containing water and alcohol. The composition may further comprise a pharmaceutically acceptable carrier.
In one embodiment of the present invention, the composition may be in the form of a powder, granules, liquid, gel or paste.
In an embodiment of the present invention, the time of the solvent extraction is 0.5 to 3 hours, the temperature of the solvent extraction is 50 to 100 ℃, and the concentration of the longan shell extract is more than 4 mg/ml.
The longan shell extract of the present invention can inhibit lipogenesis, reduce GPT index and protect liver.
The following description of the present invention is provided in connection with the accompanying drawings, which are included to illustrate and not to limit the scope of the present invention, and it will be understood by those skilled in the art that various changes and modifications may be made therein without departing from the spirit and scope of the invention, and it is intended to cover all modifications and equivalents as may fall within the true spirit and scope of the invention.
Drawings
FIG. 1 shows evaluation of oil red quantification of longan hull extract for inhibiting lipogenesis in HepG2 cells treated with 4mg/mL longan hull extract;
FIG. 2 is a graph of oil red staining for inhibition of adipogenesis by longan hull extracts evaluated from HepG2 cells treated with 4mg/mL longan hull extracts;
FIG. 3 shows HepG2 cells after pretreatment with 4mg/mL longan shell extract and H2O2Analyzing the GPT content of the induced damage;
FIG. 4 shows HepG2 cells after pretreatment with 4mg/mL longan shell extract and H2O2Fluorescence staining pattern of injured cells;
FIG. 5 shows HepG2 cells after pretreatment with 4mg/mL longan shell extract and H2O2Analysis results of expression quantity of genes related to lipogenesis synthesis induced by injury.
Detailed Description
The invention provides an application of longan shell extract in preparing a composition for regulating and controlling SREBP-1c, ACC and SCD-1 proteins, the longan shell extract is obtained by extracting longan shells at different temperatures, extracting the longan shells by using a solvent and centrifuging, and can be used for inhibiting fatty liver formation, preventing liver injury, reducing GTP (aspartate pyruvate transaminase) value and reducing liver fat synthesis related genes.
The longan shell extract of the invention is obtained by an extraction method, which comprises the following steps: (a) extracting longan shell with an extraction solvent, wherein the extraction is carried out at 50-100 ℃, and the volume ratio of the extraction solvent to the longan shell is 5-20: 1-5; (b) centrifuging the obtained product; (c) filtering the centrifuged supernatant to obtain a filtrate; (d) carrying out reduced pressure concentration on the filtrate at 45-70 ℃ to obtain a concentrated product; and (e) spray drying the concentrated product, wherein the extraction solvent is water, alcohols, aqueous alcohols, or a combination thereof.
The present invention also provides a composition for modulating SREBP-1c and ACC and SCD-1 protein, comprising an effective amount of an extract of longan shell and a pharmaceutically acceptable carrier, wherein the composition is in the form of powder, granule, liquid, gel or paste, and the dosage form is provided in the form of food, drink, pharmaceutical product, reagent or nutritional supplement.
The detailed extraction method of the longan shell extract of the present invention and the test experiments of the longan shell extract for inhibiting fatty liver formation, preventing liver damage, reducing GTP value and reducing genes related to liver fat synthesis will be described in detail below to confirm the effects of the longan shell extract.
The longan shell extraction process comprises the following steps:
1. after the longan shell is cleaned and sundries are removed, the longan shell is dried for subsequent extraction.
2. And (3) crushing the fragments with the crushing distance of about 0.2-0.5 cm.
3. Taking the crushed longan shell, and mixing the extraction solvent and the crushed longan shell according to the volume ratio of 5-20: 1-5, and extracting in a solvent at 50-100 ℃ for 0.5-3 hours.
4. And cooling to room temperature.
5. Centrifuging at 5000r.p.m. for 10 min, and removing residue.
6. Filtered through a 400 mesh screen.
7. Concentrating under reduced pressure at 45-70 deg.C.
Fatty liver cell assay:
experimental materials:
(1) cell line HepG2(ATCC, HB-8065)
(2) Culture solution Dulbecco's Modified Eagle culture solution (Gibco, Cat.12100-038) containing 10% fetal bovine serum (Gibco, Cat.10438-026) and 1% penicillin/streptomycin (Gibco, Cat.15140-122)
(3) Free fatty acid (Oleic acid; Sigma, # O1008)
(4) Bovine serum albumin (Bio Basic Inc., Cat. AD0023)
(5) Oil red O (Sigma, Cat. O0625)
(6) Isopropanol (Echo chemical, PH-3101)
(7) 10% Formaldehyde (Echo chemical, Cat. TG1794-4-0000-72NI)
(8) Phosphate buffered saline (Gibco, Cat.14200-075)
The experimental steps are as follows:
(1) in six-well plates at 5X 10 per well5Individual cells were implanted at a density and incubated overnight at 37 ℃.
(2) Cells were treated with 4mg/mL of longan shell extract in a culture medium containing 2% fetal bovine serum for 24 hours.
(3) A culture medium containing OA-BSA conjugate, 2% fetal bovine serum, and 1% penicillin/streptomycin was prepared.
(4) Removing the original culture solution and replacing the culture solution containing OA-BSA conjugate prepared in the step (3). Adding the longan shell extract with proper concentration, and treating for 24 hours.
(5) Remove the broth and wash 2 times with 1X phosphate buffer.
(6) Cells were fixed with 10% formaldehyde for 30 min.
(7) Wash 2 times with 1X phosphate buffer and rinse 15 seconds with 50% isopropanol.
(8) Stain with oil red O in 60% isopropanol for 1 hour.
Quantification was done by microscopic observation and staining was solubilized with 100% isopropanol. Statistics were student's t-test using Excel software.
Experimental results fig. 1 shows that HepG2 cells treated with 4mg/mL longan shell extract can inhibit lipogenesis by 40.5%, and can effectively reduce lipogenesis amount of HepG2 cells, thus confirming that longan shell extract can effectively inhibit HepG2 cells from forming fat. Also, as shown in the stained photograph of fig. 2, the extract of longan shell can inhibit fat to achieve the effect of preventing fatty liver.
Gpt (alt) assay for liver injury cells:
experimental materials:
(1) cell line: HepG2(ATCC, HB-8065)
(2) Culture solution: dulbecco's Modified Eagle Medium (Gibco, Cat.12100-038) containing 10% fetal bovine serum (Gibco, Cat.10438-026) and 1% penicillin/streptomycin (Gibco, Cat.15140-122)
(3) An ELISA kit (USCN., Cat. SEA207Hu) for Alanine Aminotransferase (ALanine Aminotranferase, ALT) phosphate buffer (Gibco, Cat.14200-075)
(4) ELISA reader (BioTek)
The experimental steps are as follows:
(1) at 2X 10 per well4The density of individual cells was plated in 24-well culture plates and incubated overnight at 37 ℃.
(2) In the presence of H2O2Prior to the treatment, the cells were pretreated in culture medium for 24 hours at 4mg/ml of longan shell extract.
(3) After 24 hours of pretreatment, H was added2O2The treatment was carried out at a concentration of 500. mu.M for 6 hours.
(4) Staining the cells with propidium iodide (propidium iodide) staining solution (1:250) and Annexin V (1:250) in Annexin V bonding buffer for 15-30 min.
(5) Cells were stained with Hoechst 33342(1:20000) for 3 minutes.
(6) Cells were washed 2 times with phosphate buffer.
(7) Observed with a fluorescence microscope.
The results are shown in FIG. 3, which shows HepG2 cells passing through H2O2After injury, the GPT index is increased dramatically, however, the GPT index can be effectively reduced by 90% by the group pretreated by the longan shell extract with the concentration of 4 mg/ml.
Gene level discussion of longan shell extract
Experimental materials:
(1) cell line: HepG2(ATCC, HB-8065)
(2) Culture solution Dulbecco's Modified Eagle culture solution (Gibco, Cat.12100-038) containing 10% fetal bovine serum (Gibco, Cat.10438-026) and 1% penicillin/streptomycin (Gibco, Cat.15140-122)
(3) An ELISA kit for Alanine Aminotransferase (ALaninotransferase, ALT) (USCN., Cat. SEA207Hu)
(4) Phosphate buffer solution (Gibco, Cat.14200-075)
(5) ELISA reader (BioTek)
The experimental steps are as follows:
(1) in six-well plates at 1X 10 per well5The cells, 1mL of culture medium were implanted and incubated overnight at 37 ℃.
(2) Treating with 2mg/ml or 4mg/ml longan shell extract for 24 hours.
(3) Collecting the stimulated cells.
(4) Collecting RNA from the cells using an RNA extraction kit.
(5) Reverse transcribing the RNA with reverse transcriptase to obtain cDNA.
(6) qPCR was performed using KAPA SYBR FAST using ABI Step One Plus to quantify the target gene.
(7) Relative amount of gene expression in 2-ΔΔCtAnd (4) determining the method.
Statistical analysis was performed with student's t-test in Excel software (＊: P < 0.05; ＊＊: P <0.01)
The results are shown in FIG. 4, which shows HepG2 cells passing through H2O2The longan shell extract can improve the survival rate of cells. This indicates that the longan shell extract can protect the liver from damage caused by external stimuli. As discussed in the gene level of longan shell extract, when liver cells are externally stimulated, SREBP-1c transcription factors are generated, and the transcription factors can enable the cells to generate proteins ACC and SCD-1 related to synthetic lipid. As shown in FIG. 5, the longan shell extract showed significant differences in the reduction of SREBP-1c by 78%, the reduction of ACC by 82% and the reduction of SCD-1 by 33%, and all of the 3 proteins increased the intracellular fat (trigyceries) in the liver cells, resulting in fatty liver.
In conclusion, the longan shell extract of the present invention has the efficacy of regulating SREBP-1c, ACC and SCD-1 proteins, and can be used as food, beverage, drug, reagent or nutritional supplement for regulating SREBP-1c, ACC and SCD-1 proteins, etc. regardless of cell experiments and gene expression.
Therefore, the longan shell extract provided by the invention can effectively inhibit the formation of fatty liver, prevent liver injury, reduce GTP value and reduce genes related to liver lipogenesis synthesis so as to achieve the effect of protecting liver.
Claims (10)
1. Use of a longan shell extract for preparing a composition for reducing the expression of SREBP-1c, ACC and SCD-1 proteins, wherein the longan shell extract is obtained by extracting longan shells with a solvent selected from the group consisting of water, alcohol and aqueous alcohol.
2. The use of claim 1, wherein said composition further comprises a pharmaceutically acceptable carrier.
3. Use according to claim 1, wherein the composition is in the form of a powder, granules, liquid, gel or paste.
4. The use according to claim 1, wherein the concentration of the extract of longan shell is 4mg/ml or more.
5. The use as claimed in claim 1, wherein the volume ratio of the solvent to longan shell in the extraction is 5-20: 1 to 5.
6. Use according to claim 1, wherein the solvent is extracted for a period of 0.5 to 3 hours.
7. Use according to claim 1, wherein the solvent has an extraction temperature of 50 to 100 ℃.
8. Use according to any one of claims 1 to 7, wherein the longan shell extract inhibits lipogenesis.
9. The application of the longan shell extract in preparing the composition for protecting the liver is characterized in that the longan shell extract is obtained by extracting longan shells with water, alcohol or a solvent containing water and alcohol.
10. The use as claimed in claim 9, wherein the extract of longan hull reduces the GPT index.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115025078A (en) * | 2022-05-24 | 2022-09-09 | 长江师范学院 | Application of longan pomace polyphenol extract in preparation of medicine for preventing and treating non-alcoholic steatohepatitis |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108066468A (en) * | 2016-11-17 | 2018-05-25 | 百岳特生物科技（上海）有限公司 | Longan shell extract and its use |
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108066468A (en) * | 2016-11-17 | 2018-05-25 | 百岳特生物科技（上海）有限公司 | Longan shell extract and its use |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115025078A (en) * | 2022-05-24 | 2022-09-09 | 长江师范学院 | Application of longan pomace polyphenol extract in preparation of medicine for preventing and treating non-alcoholic steatohepatitis |
| CN115025078B (en) * | 2022-05-24 | 2024-02-23 | 长江师范学院 | Application of longan pomace polyphenol extract in preparation of medicine for preventing and treating non-alcoholic steatohepatitis |
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| TA01 | Transfer of patent application right |
Effective date of registration: 20200619 Address after: No. 989, Jinge Road, Jinshan Industrial Zone, Jinshan District, Shanghai Applicant after: TCI Co.,Ltd. Address before: Taipei City, Taiwan, China Applicant before: TCI Co.,Ltd. |
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Application publication date: 20200214 |
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