阅读说明：本技术 一种细柱五加叶总提物在制备降血糖药物中的应用 (Application of acanthopanax gracilistylus leaf total extract in preparation of hypoglycemic drugs ) 是由 鲁曼霞 刘向前 杨阳 于 2020-05-21 设计创作，主要内容包括：本发明公开了一种细柱五加叶总提物在制备降血糖药物中的应用,所述细柱五加叶总提物的制备过程为：将细柱五加叶置于多功能提取罐,以水或甲醇作为溶剂,在20～80℃提取1～3次,合并后减压回收溶剂后即得总提物。本发明创新的发现,细柱五加叶总提物具有显著的降血糖活性,基于此,本发明提供了一种细柱五加叶总提物在制备降血糖药物中的全新用途,在制备成防治糖尿病的药物方面具有广阔的应用前景。(The invention discloses an application of acanthopanax gracilistylus leaf total extract in preparing a hypoglycemic medicament, wherein the process for preparing the acanthopanax gracilistylus leaf total extract comprises the following steps: placing the acanthopanax gracilistylus leaves in a multifunctional extraction tank, extracting for 1-3 times at 20-80 ℃ by using water or methanol as a solvent, combining, and recovering the solvent under reduced pressure to obtain a total extract. The invention innovatively discovers that the acanthopanax gracilistylus leaf total extract has obvious hypoglycemic activity, and on the basis, the invention provides a brand new application of the acanthopanax gracilistylus leaf total extract in preparing hypoglycemic drugs, and the acanthopanax gracilistylus leaf total extract has wide application prospect in preparing drugs for preventing and treating diabetes.)

1. The application of the acanthopanax gracilistylus leaf total extract in preparing the hypoglycemic drug is characterized in that the acanthopanax gracilistylus leaf total extract is prepared by the following steps: placing the acanthopanax gracilistylus leaves in a multifunctional extraction tank, extracting for 1-3 times at 20-80 ℃ by using water or methanol as a solvent, combining, and recovering the solvent under reduced pressure to obtain a total extract.

2. The use of the total extract of leaves of Acanthopanax gracilistylus W.W. Smith as claimed in claim 1 for preparing hypoglycemic agent, wherein: the application of the acanthopanax senticosus leaf total extract in preparing a medicament for preventing and treating diabetes mellitus.

3. The use of the total extract of leaves of Acanthopanax gracilistylus W.W. Smith as claimed in claim 2 for preparing hypoglycemic agent, wherein: the medicament for preventing and treating diabetes is a medicament capable of inhibiting activities of alpha-glucosidase, alpha-amylase and PTP 1B.

4. The use of the total extract of leaves of Acanthopanax gracilistylus W.W. Smith as claimed in claim 2 for preparing hypoglycemic agent, wherein: the medicament for preventing and treating diabetes is a medicament capable of promoting insulin secretion under the stimulation of glucose.

5. The use of the total extract of leaves of Acanthopanax gracilistylus W.W. Smith as claimed in claim 2 for preparing hypoglycemic agent, wherein: the medicine for preventing and treating diabetes is a medicine capable of partially restoring cell activity in RIN-m5F cells subjected to cell apoptosis treatment by cytokines IL-1 beta and IFN-gamma induction RIN-m5F in a concentration-dependent mode and reducing Caspase-3 activity in RIN-m5F cells subjected to cell apoptosis treatment by the cytokines IL-1 beta and IFN-gamma induction RIN-m5F in a concentration-dependent mode.

6. The use of the total extract of leaves of Acanthopanax gracilistylus W.W. Smith as claimed in claim 2 for preparing hypoglycemic agent, wherein: the medicine for preventing and treating diabetes mellitus is a medicine capable of regulating the levels of NO and ROS in RIN-m5F cells subjected to apoptosis treatment by cytokines IL-1 beta and IFN-gamma in a concentration-dependent manner and RIN-m5F cells.

7. The use of the total extract of leaves of Acanthopanax gracilistylus W.W. Woodlings as claimed in any of claims 1 to 6 for the preparation of hypoglycemic agents, characterized in that: in the medicament for preventing and treating diabetes, the mass percentage of the acanthopanax gracilistylus leaf total extract is 0.1-100%.

8. The use of the total extract of leaves of Acanthopanax gracilistylus W.W. Woodlings as claimed in any of claims 1 to 6 for the preparation of hypoglycemic agents, characterized in that: the medicament for preventing and treating diabetes also comprises pharmaceutically acceptable auxiliary materials or carriers.

9. The use of the total extract of leaves of Acanthopanax gracilistylus W.W. Woodlings as claimed in any of claims 1 to 6 for the preparation of hypoglycemic agents, characterized in that: the dosage form of the medicament for preventing and treating the diabetes mellitus is tablets, granules, capsules, dripping pills, oral liquid or injection.

Technical Field

The invention belongs to the field of medicine application, and particularly relates to application of a acanthopanax gracilistylus leaf total extract in preparation of a hypoglycemic medicine.

Background

At present, diabetes is an endocrine metabolic disease seriously threatening human health, and the incidence rate is on the rising trend year by year. Has become a common and frequently encountered disease in many countries worldwide, with mortality rates third after cancer, cardiovascular. The number of diabetes patients in China is the first global, and about 95 percent of the diabetes patients are type II diabetes patients. Type II diabetes is non-insulin dependent diabetes mellitus, the etiology and pathogenesis of which are complex, and islet beta cell apoptosis and Insulin Resistance (IR) are the core of the study of type II diabetes. The islet beta cell apoptosis is not only a direct cause of the onset of type II diabetes, but also plays an important role in the occurrence and development of type II diabetes. Nuclear factor- κ B (NF- κ B) is involved in the immunomodulation of gene transcription mediating inflammation and various pathophysiological processes. Recent studies have shown that NF- κ B is closely associated with diabetes and obesity. Domestic and foreign researches show that the inflammation also participates in insulin resistance and pancreatic islet beta cell function damage, thereby causing the occurrence of diabetes. The inflammatory cytokine interleukin IL-1 β is a key cytokine that causes apoptosis of islet β cells in type I and type II diabetes. IL-1 beta acts on islet beta cells, influences the synthesis and secretion of insulin, and can generate a large amount of NO to cause the apoptosis of the islet beta cells by inducing the expression Increase of Nitric Oxide Synthase (iNOS) in the cells. IL-1 beta can also up-regulate beta cell transmembrane protein Fas expression by activating NF-kappa B, and then activate pro-caspase-8 to crack the protein to generate active caspase-8, so as to activate caspase-3 and finally cause islet beta cell apoptosis. Meanwhile, the oxidative stress can induce the islet beta cell apoptosis through an NF-kB-iNOS-NO signal channel. Oxidative damage is mainly caused by Reactive Oxygen Species (ROS), and among various pathways for oxidative damage to induce β cell apoptosis, activation of NF- κ B and activation of Inducible Nitric Oxide Synthase (iNOS) have been the major concerns.

Acanthopanax gracilistylus W.W.W.W.W.W.W.W.W.W.W.Wright has the functions of resisting inflammation, resisting rejection, raising immunity, promoting nucleic acid formation, resisting fatigue, etc. The acanthopanax gracilistylus leaf is less researched, and no relevant report on the aspect of reducing blood sugar is found at present.

Disclosure of Invention

The invention innovatively discovers that the acanthopanax gracilistylus total extract has obvious hypoglycemic activity. Based on the above, the invention aims to provide a brand new application of the acanthopanax gracilistylus leaf total extract in preparing a hypoglycemic medicament.

In order to achieve the technical purpose, the invention adopts the following technical scheme:

the application of the acanthopanax gracilistylus leaf total extract in preparing the hypoglycemic drug comprises the following steps: placing the acanthopanax gracilistylus leaves in a multifunctional extraction tank, extracting for 1-3 times at 20-80 ℃ by using water or methanol as a solvent, combining, and recovering the solvent under reduced pressure to obtain a total extract.

Preferably, the acanthopanax gracilistylus leaf total extract is applied to preparation of a medicament for preventing and treating diabetes.

The medicament for preventing and treating diabetes is a medicament capable of inhibiting activities of alpha-glucosidase, alpha-amylase and PTP 1B.

The medicament for preventing and treating diabetes is a medicament capable of promoting insulin secretion under the stimulation of glucose.

The medicine for preventing and treating diabetes is a medicine capable of partially recovering cell viability in RIN-m5F cells subjected to apoptosis treatment by cytokines IL-1 beta and IFN-gamma in a concentration-dependent manner and RIN-m 5F.

The medicine for preventing and treating diabetes is a medicine capable of reducing Caspase-3 activity in RIN-m5F cells subjected to apoptosis treatment by using cytokines IL-1 beta and IFN-gamma to induce RIN-m5F cells in a concentration-dependent manner.

The medicine for preventing and treating diabetes mellitus is a medicine capable of regulating the NO level in RIN-m5F cells subjected to apoptosis treatment by cytokines IL-1 beta and IFN-gamma in a concentration-dependent manner, wherein RIN-m5F cells are induced to undergo apoptosis.

The medicine for preventing and treating diabetes mellitus is a medicine capable of regulating the ROS level in RIN-m5F cells subjected to apoptosis treatment by cytokines IL-1 beta and IFN-gamma in a concentration-dependent mode and RIN-m5F cells.

Preferably, in the medicament for preventing and treating diabetes, the mass percentage of the total extract of the acanthopanax gracilistylus leaves is 0.1-100%.

Preferably, the medicament for preventing and treating diabetes further comprises a pharmaceutically acceptable adjuvant or carrier.

The application of the invention can be combined with the prior pharmaceutically acceptable auxiliary materials or carriers to prepare any pharmaceutically acceptable dosage form.

Preferably, the dosage form of the medicament for preventing and treating diabetes is tablets, granules, capsules, dripping pills, oral liquid or injections.

Has the advantages that:

(1) the acanthopanax gracilistylus leaf total extract has obvious effect of inhibiting alpha-glucosidase, alpha-amylase and PTP 1B.

(2) The acanthopanax gracilistylus leaf total extract can effectively promote insulin secretion under the stimulation of glucose.

(3) The total extract of leaves of Acanthopanax gracilistylus W.W.W.W.W.W.W.W.W.W.can partially restore cell viability in RIN-m5F cells treated with cytokine IL-1 beta and IFN-gamma-induced apoptosis of RIN-m5F in a concentration-dependent manner, and can restore cell viability to the level of the group not treated with the cytokine.

(4) The total extract of the acanthopanax gracilistylus leaf can regulate the NO level in RIN-m5F cells subjected to apoptosis treatment of RIN-m5F cells induced by cytokines IL-1 beta and IFN-gamma in a concentration-dependent mode.

(5) The total extract of the acanthopanax gracilistylus leaves can reduce the Caspase-3 activity in RIN-m5F cells subjected to apoptosis treatment of RIN-m5F cells induced by cell factors IL-1 beta and IFN-gamma in a concentration-dependent mode.

(6) The acanthopanax gracilistylus leaf total extract can regulate the ROS level in RIN-m5F cells subjected to apoptosis treatment of RIN-m5F cells induced by cytokines IL-1 beta and IFN-gamma in a concentration-dependent mode.

In conclusion, the acanthopanax gracilistylus leaf total extract has obvious hypoglycemic activity and has wide application prospect in the aspect of preparing medicines for preventing and treating diabetes.

Drawings

FIG. 1 is a graph showing the effect of the total extract of leaves of Acanthopanax gracilistylus W.W.W.W.W.W.W.W.W.W.W.W.in example 2 (methanol extract) on the activity of RIN-m5F cells.

FIG. 2 is a graph showing the results of the effect of the total extract of leaves of Acanthopanax gracilistylus W.W. (aqueous extract WA and methanol extract ME) on glucose-stimulated insulin secretion in example 3; RIN-m5F cells were cultured at basal (4mM) or stimulated (20mM) glucose concentration in the presence of Acanthopanax gracilistylus leaf total extract; p <0.05, relative to vehicle-treated control; bars represent SEM (n ═ 3).

FIG. 3 is a graph showing the effect of the total extract of leaves of Acanthopanax gracilistylus W.W.W.W.W.W.W.W.W.in example 4 (methanol extract) on cytokine-induced cell death; wherein-represents untreated, + represents treated, P <0.05 compared to control of blank treatment; # P <0.01 compared to cytokine treated group; bars represent SEM (n ═ 3).

FIG. 4 is a graph showing the effect of the total extract of leaves of Acanthopanax gracilistylus W.W.W.W.W.W.W.W.W.W.W.in example 5 (methanol extract) on the cytokine-induced production of NO in RIN-m5F cells; wherein-represents untreated, + represents treated, P <0.05 compared to control of blank treatment; # P <0.01 compared to cytokine treated group; bars represent SEM (n ═ 3).

FIG. 5 is a graph showing the effect of the total extract of leaves of Acanthopanax gracilistylus W.W.W.W.W.W.W.W.W.W.W.in example 6 on cytokine-induced Caspase-3 activity in RIN-m5F cells; wherein-represents untreated, + represents treated, P <0.05 compared to control of blank treatment; # P <0.01 compared to cytokine treated group; bars represent SEM (n ═ 3).

FIG. 6 is a graph showing the effect of the total extract of leaves of Acanthopanax gracilistylus W.W.W.W.W.W. (methanol extract) on cytokine-induced ROS levels in RIN-m5F cells in example 7; wherein-represents untreated, + represents treated, P <0.05 compared to control of blank treatment; # P <0.01 compared to cytokine treated group; bars represent SEM (n ═ 3).

Detailed Description

The acanthopanax gracilistylus leaf is extracted by water or alcohol (methanol and/or ethanol) and concentrated to obtain an extract of the acanthopanax gracilistylus leaf, and the specific preparation process comprises the following steps: placing the acanthopanax gracilistylus leaves in a multifunctional extraction tank (common extraction equipment in the field), respectively taking water and methanol as solvents, extracting for 1-3 times at 30-80 ℃, combining, and recovering the solvents under reduced pressure to obtain a total extract (an aqueous extract or a methanol extract).

Subjecting the total extract to high performance liquid chromatography with Phe-nomenex Columbus C18110A chromatographic column (250mm × 4.60mm, 5.0 μm); the column temperature is 30 ℃; taking acetonitrile-0.2% phosphoric acid water as a mobile phase, and performing gradient elution: 0-5 min, 45% A; 45-50% of A for 5-14 min; 14-28 min, 50% -60% A; 28-40 min, 60% A; the detection wavelength is 210 nm; the flow rate was 1m L/min, and was measured simultaneously. The total extract comprises impressic acid, aconkoregenin, acontrifoside A, aconkoreoside D, aconkoreoside B, 3-O-beta-D-glucopyranosyl 3 alpha, 11 alpha-dihydroxyup-20 (29) -en-28-oic acid, 3 alpha, 11 alpha, 23-trihydroxy-lup-20(29) -en-28-oic acid and 3 alpha, 11 alpha-dihydroxyx-23-oxo-lup-20 (29) -en-28-oic acid which are detected, and the total mass percent of the total extract is not less than 5-10% of the total extract.