CN112603923A

CN112603923A - Application of phillyrin in preparation of medicine for preventing or/and treating type II diabetes

Info

Publication number: CN112603923A
Application number: CN202011560012.1A
Authority: CN
Inventors: 韩荣春; 童小慧; 易雨琪; 吕燕萍; 卢孟雅; 方志正; 俞年军
Original assignee: Anhui University of Traditional Chinese Medicine AHUTCM
Current assignee: Anhui University of Traditional Chinese Medicine AHUTCM
Priority date: 2020-12-25
Filing date: 2020-12-25
Publication date: 2021-04-06

Abstract

The invention relates to the technical field of medicine application, in particular to application of phillyrin in preparing a medicine for preventing or/and treating type II diabetes, wherein the molecular weight of the phillyrin is 534.55; meanwhile, from the economic perspective, the phillyrin and the hypoglycemic product thereof have extremely wide application space.

Description

Application of phillyrin in preparation of medicine for preventing or/and treating type II diabetes

Technical Field

The invention relates to the technical field of medicine application, in particular to application of phillyrin in preparation of a medicine for preventing or/and treating type II diabetes.

Background

Diabetes is a series of metabolic disorder syndromes with continuously high blood sugar levels as important indicators. The symptoms are usually manifested as polyuria, thirst, polyphagia, etc. If not treated for a prolonged period of time, diabetic patients can develop a variety of complications. Acute complications may include hyperosmolar hyperglycemic states, diabetic ketoacidosis, and even death. Serious long-term complications include foot ulcers, stroke, chronic kidney disease, cardiovascular disease, ocular fundus disease, nervous system disorders, and cognitive disorders.

The occurrence of diabetes can be attributed to two causes: first, the pancreas cannot secrete sufficient insulin; secondly, the responsiveness of cells in the body to insulin is reduced. Diabetes can be mainly divided into three categories: type one diabetes, type two diabetes and type gestational diabetes. During the development and development of type II diabetes, cells in the body, particularly adipocytes, fail to respond correctly to insulin, leading to the phenomenon of insulin resistance. As diabetes progresses, insufficient insulin secretion may occur simultaneously. The most common causes of this may be excessive weight and lack of exercise.

By 2019, it is estimated that 8.8% of the adults worldwide suffer from diabetes, which corresponds to a population number of 4.7 billion; of these, type II diabetes accounts for 90%. Studies have shown that type two diabetes mellitus occurs without sex differences, i.e. men and women have a comparable chance of developing the disease. Meanwhile, the incidence of diabetes still tends to rise greatly along with the improvement of living standard, the change of dietary structure and the reduction of daily life exercise amount of residents. In 2019 alone, it is estimated that diabetes directly causes death of over 400 million people, making it the seventh leading death disease worldwide. While the costs associated with diabetes treatment are statistically more than seven billion dollars globally spent in 2017. The living standard of the diabetic is greatly influenced, and meanwhile, the heavy economic burden is brought to the family of the diabetic.

The common diabetes treatment drugs can be divided into eight groups. Mainly comprises insulin and mimetics, metformin compounds, sulfonylurea drugs, glucosidase inhibitors, thiazolidinedione drug sensitizers, anisic acid derivatives, DPP-4 enzyme inhibitors, receptor agonists and the like.

The diabetes treatment drugs sold on the market at present are various in variety, but most of the drugs are biological products or artificially synthesized monomeric compounds. The disadvantages include: high requirement on storage conditions, short drug action time, frequent administration, high drug selling price, great side effect and the like.

In view of the above-mentioned drawbacks, the inventors of the present invention have finally obtained the present invention through a long period of research and practice.

Disclosure of Invention

The invention aims to solve the problems that currently marketed diabetes treatment medicines are various in varieties, high in storage condition requirement, short in drug effect action time, high in medicine selling price, large in side effect and the like, and frequent administration is needed, and provides application of phillyrin in preparation of medicines for preventing or/and treating type II diabetes.

In order to realize the purpose, the invention discloses an application of phillyrin in preparing a medicament for preventing or/and treating type II diabetes, wherein the chemical structural formula of the phillyrin is as follows:

the phillyrin can improve chronic inflammation state of visceral fat in type II diabetes, and has positive effect on diabetes treatment by improving visceral fat STAT3-SOCS3 pathway abnormality in type II diabetes.

Compared with the prior art, the invention has the beneficial effects that: the monomer compound phillyrin extracted from traditional Chinese medicine forsythia is used as the only effective component for treating mouse diabetes, and a good effect is achieved; phillyrin produces a positive effect on the treatment of diabetes by ameliorating the abnormalities in the visceral fat STAT3-SOCS3 pathway in type ii diabetic state; the fructus forsythiae in China is rich in resources, and the fructus forsythiae is high in phillyrin content and low in price; the phillyrin is used for treating diabetes, so that the blood sugar of a mouse can be obviously reduced, and the living quality of experimental animals can be improved; meanwhile, from the economic perspective, the phillyrin and the hypoglycemic product thereof have extremely wide application space.

Drawings

FIG. 1 is a comparison of random blood glucose levels of various groups of experimental mice in an example of the present invention;

FIG. 2 is a comparison of fasting blood glucose levels of experimental mice in each group according to the example of the present invention;

FIG. 3 shows the results of oral glucose tolerance experiments in mice according to the present invention.

FIG. 4 shows the results of experiments on the weights of diabetic mice in the phillyrin middle and high dose groups according to the present invention;

FIG. 5 shows the effect of phillyrin intermediate and high dose groups on the expression level of inflammatory factors associated with diabetic mice in an example of the present invention;

FIG. 6 is a graph of the effect of phillyrin intermediate and high dose groups on the metabolic pathways associated with diabetic mice in an example of the present invention.

Detailed Description

The above and further features and advantages of the present invention are described in more detail below with reference to the accompanying drawings.

Hypoglycemic activity of phillyrin on type II diabetic mice

The experimental animals selected SPF grade C57BL/6J male 4 weeks old, 60 in total. The breeding condition is 24 ℃; 12 hours light, 12 hours dark; can be freely eaten and drunk. Wherein 10 mice in the normal control group are fed with the maintenance feed to the experimental mice; the remaining 50 mice (experimental group) were given 60% fat-energized high-fat diet.

Step 1: model making of type II diabetes mouse

After normal feeding for 8 weeks, the experimental animals were each subjected to a single intraperitoneal injection of streptozotocin (30mg/kg, citrate buffer pH 4.4); the mice in the normal control group were injected with citrate buffer (pH 4.4), respectively. All experimental mice were bled from the tail vein after 6h fasting (free drinking) with streptozotocin or citrate buffer injection for 18h, 42h and 90h, respectively, to test blood glucose levels with a rotz reasonger glucometer-i.e. fasting blood glucose levels were tested for all test mice at 24h, 48h and 96 h. The model making of the diabetic mice is judged to be successful when the fasting blood sugar level is higher than 13.3 mmol/L.

Step 2: hypoglycemic activity of phillyrin on type II diabetes mice

The mice in the experimental group (38 mice in total) with successful diabetes modeling are divided into a high-fat model group (9 mice), a phillyrin high-dose group (10 mice), a phillyrin medium-dose group (9 mice) and a phillyrin low-dose group (10 mice), and the mice in each experimental group are continuously fed with high-fat feed. The normal control group was continued to feed the experimental rats with the maintenance feed.

Phillyrin (purity > 98%, confirmed by high performance liquid chromatography) was suspended in 1% sodium carboxymethylcellulose (CMC-Na) solution, and the high, medium and low dose groups of phillyrin were administered by intragastric gavage for 4 weeks at the following doses, respectively: 50mg/kg,25mg/kg,12.5 mg/kg. The normal control group and the high fat model group were gavaged with a 1% sodium carboxymethylcellulose (CMC-Na) solution for 4 weeks.

After 4 weeks of gavage, randomized glycemia was measured in mice including a normal control group (n-8) and each experimental group (high-fat model group n-7, forsythia suspensa low-dose group n-8, forsythia suspensa medium-dose group n-7, and forsythia suspensa high-dose group n-8) (fig. 1). Statistical analysis was performed by T-test, and as can be seen from fig. 1, the group of high lipid models (T2DM) was significantly different (p <0.0001) compared to the normal control group (Ctrl); the T2DM group is significantly different from the phillyrin middle dose group (Med) (p < 0.0001); the T2DM group was significantly different from the phillyrin High dose group (High) (p < 0.0001). Therefore, the forsythin has obvious effect on reducing the random blood sugar level of the diabetic mice.

Meanwhile, fasting blood glucose of each group of experimental animals was measured and statistically analyzed by T-test, as shown in fig. 2, the group of high fat model (T2DM) was significantly different (p <0.0001) compared to the normal control group (Ctrl); the T2DM group is significantly different from the phillyrin middle dose group (Med) (p < 0.001); the T2DM group was significantly different from the phillyrin High dose group (High) (p < 0.0001). It can be seen that phillyrin has significant efficacy in lowering fasting blood glucose levels in diabetic mice.

Glucose tolerance effect of forsythin and II on type II diabetic mice

Subsequently, glucose tolerance test was performed on each group of mice by the following method: after fasting the mice overnight (free drinking water), the glucose solution was gavaged at 2g/kg body weight, followed by tail vein bleeding and blood glucose levels measured at 0min, 15min, 30min, 60min and 120min after the glucose gavage and area under the curve calculated (fig. 3). According to the results of oral glucose tolerance experiments of mice (figure 3), the area under the curve is analyzed statistically by T test, and the significant difference (p <0.0001) is found in the high-fat model group (T2DM) compared with the normal control group (Ctrl); the T2DM group is significantly different from the phillyrin middle dose group (Med) (p < 0.01); the T2DM group was significantly different from the phillyrin High dose group (High) (p < 0.001). It is inferred that the phillyrin has a significant effect of increasing the glucose tolerance level when administered orally at medium and high doses and thus exerts an effect of treating diabetes.

Third, the Effect of Forsythiaside on the body weight of type II diabetic mice

Before sacrifice, the weight of each group of mice was weighed. The results show that the weight of the type II diabetic mice can be obviously reduced by the medium and high dosage of the medicament. It was concluded that the medium and high doses of phillyrin, when administered orally, improved the body weight of obesity-related type ii diabetic mice (figure 4). As shown in fig. 4, the group of high lipid model (T2DM) was significantly different (p <0.001) compared to the normal control group (Ctrl); compared with the phillyrin middle dose group (Med) and the phillyrin High dose group (High), the T2DM group has significant differences (p is less than 0.001), and therefore, the phillyrin middle dose and the phillyrin High dose have significant effects on reducing the body weight of the type II diabetic mice when orally administered.

Effect of forsythin on visceral adipose tissue inflammation of type II diabetic mice

Mice were sacrificed by cervical dislocation 4 weeks after administration, followed by isolation of epididymal adipose tissue and snap-freezing with liquid nitrogen. And extracting the RNA of the epididymis adipose tissue by Trizol, and detecting the expression level of the related inflammatory genes by qRT-PCR after reverse transcription. The levels of inflammatory factors of tnf alpha, il6 and mcp1 were experimentally examined, and as a result, the visceral adipose tissue inflammation level of mice in the T2MD group was found to be significantly increased compared with that of the normal control group. In contrast, the level of inflammation in adipose tissues was significantly reduced in the phillyrin medium-dose and high-dose groups compared to the T2DM group (fig. 5). It is concluded that the intermediate and high doses of phillyrin, when administered orally, improve the chronic inflammatory state of visceral fat in type II diabetic condition.

Fifthly, the effect of phillyrin on visceral fat STAT3-SOCS3 pathway of type II diabetic mice

Mice were sacrificed by cervical dislocation 4 weeks after administration, followed by isolation of epididymal adipose tissue and snap-freezing with liquid nitrogen. And extracting the RNA of the epididymis adipose tissue by Trizol, and detecting the expression level of the related inflammatory genes by qRT-PCR after reverse transcription. The results are shown in fig. 6, and the middle and high phillyrin dose groups can significantly reverse the expression of SOCS3 gene in visceral fat of mice in type ii diabetes models. It is inferred that the intermediate dose and the high dose of forsythin, when administered orally, can improve the abnormality of visceral fat STAT3-SOCS3 pathway in type II diabetic state, thereby having a positive effect on the treatment of diabetes.

The foregoing is merely a preferred embodiment of the invention, which is intended to be illustrative and not limiting. It will be understood by those skilled in the art that various changes, modifications and equivalents may be made therein without departing from the spirit and scope of the invention as defined in the appended claims.

Claims

1. Application of phillyrin in preparing medicine for preventing and/or treating type II diabetes is provided.

2. The use of forsythin according to claim 1 in the preparation of a medicament for the prevention or/and treatment of type ii diabetes, wherein forsythin has the following chemical formula:

3. the use of forsythin according to claim 1 in the manufacture of a medicament for the prevention or/and treatment of type ii diabetes, wherein forsythin is used in the manufacture of a medicament for the amelioration of chronic inflammation of visceral fat in the type ii diabetic state.

4. The use of phillyrin in the preparation of a medicament for the prevention or/and treatment of type ii diabetes according to claim 1, wherein phillyrin is used in the preparation of a medicament for reversing the expression of SOCS3 gene in visceral fat in type ii diabetes.