CN109893655B

CN109893655B - Application of miR-327 inhibitor and/or FGF10 promoter in medicine for preventing and/or treating diabetes

Info

Publication number: CN109893655B
Application number: CN201711310791.8A
Authority: CN
Inventors: 杨云龙; 曹义海
Original assignee: Yihui Technology Shenzhen Co ltd
Current assignee: Yihui Technology Shenzhen Co ltd
Priority date: 2017-12-11
Filing date: 2017-12-11
Publication date: 2021-07-20
Anticipated expiration: 2037-12-11
Also published as: CN109893655A

Abstract

The invention provides a new application of a miR-327 inhibitor and/or FGF10 promoter, in particular to an application in preparing a medicament for preventing and/or treating diabetes. The miR-327 inhibitor and the FGF10 promoter can promote the increase of FGF10 in a stromal cell component of adipose tissues, promote an adipocyte precursor of the stromal cell component to be differentiated into brown adipocytes, promote blood sugar consumption, improve insulin sensitivity and achieve the purpose of preventing and/or treating diabetes. The invention also provides a medicament for preventing and/or treating diabetes, which comprises one or more of miR-327 inhibitor and FGF10 promoter and a pharmaceutically acceptable carrier.

Description

Application of miR-327 inhibitor and/or FGF10 promoter in medicine for preventing and/or treating diabetes

Technical Field

The invention relates to the technical field of medicines, in particular to application of a miR-327 inhibitor and/or FGF10 promoter in medicines for preventing and/or treating diabetes.

Background

Diabetes is a group of metabolic diseases characterized by hyperglycemia. Hyperglycemia is caused by a defect in insulin secretion or an impaired biological action, or both. Diabetes is generally caused by genetic or environmental factors, with hyperphagia, reduced physical activity and obesity being the most prominent environmental factors in type ii diabetes. Over time, diabetes can damage the heart, blood vessels, eyes, kidneys, and nerves. The risk of heart attack and stroke in diabetic adults rises two to three times; diabetic retinopathy, the major cause of blindness, is the result of a long-term cumulative impairment of the small vessels of the retina, and 2.6% of blindness can be attributed to diabetes; diabetes is the leading cause of renal failure. Therefore, prevention and treatment of diabetes are particularly important for self-health.

Currently, there are many methods for the prevention and treatment of diabetes, including diet control, moderate exercise, drug therapy, and the like. Wherein, the curative effect of the medicine is more obvious than that of other methods. However, most of commercially available drugs for preventing and treating diabetes control diabetes based on the principles of stimulating insulin secretion, reducing glucose absorption in the gastrointestinal tract, and the like, but long-term use of these drugs damages organs such as liver and the like, which is not good for health. Therefore, there is a need for a drug for the prevention and treatment of diabetes without compromising physical health.

Disclosure of Invention

In order to solve the problems, the invention provides application of a miR-327 inhibitor and/or FGF10 promoter in preparation of a medicament for preventing and/or treating diabetes. The miR-327 inhibitor and the FGF10 promoter can promote the increase of FGF10, promote the differentiation of fat cell precursors to brown fat cells, promote the consumption of blood sugar, improve the sensitivity of an organism to insulin and achieve the aim of preventing and/or treating diabetes.

In a first aspect, the invention provides an application of a miR-327 inhibitor and/or FGF10 promoter in preparation of a medicament for preventing and/or treating diabetes.

The miR-327 inhibitor is used for targeting miR-327 in Stromal Vascular Fraction (SVF) in adipose tissue, reduces expression of miR-327, promotes increase of fibroblast growth factor 10(FGF10) which is a target of miR-327 in non-adipose cells (namely various cells in the stromal vascular fraction), promotes differentiation of adipose cell precursors in the stromal vascular fraction to brown adipose cells, increases the metabolic rate of an organism, promotes blood glucose consumption, and improves insulin sensitivity. Thereby achieving the effect of preventing and/or treating diabetes.

The FGF10 promoter can promote the increase of FGF10 in fat matrix vascular components (SVF), promote the differentiation of fat cell precursors in the SVF to brown fat cells, increase the body metabolic rate, promote blood sugar consumption and improve insulin sensitivity. Can also be used for preventing and/or treating diabetes.

Optionally, the medicament for preventing and/or treating diabetes comprises a miR-327 inhibitor and an FGF10 promoter at the same time. Under the condition of the two components existing at the same time, the function of synergistically promoting the insulin sensitivity can be achieved.

In the present invention, miR-327 is a micro RNA (MicroRNA), a short non-coding RNA, that regulates protein expression post-transcriptionally.

Optionally, the miR-327 inhibitor comprises one or more of a chemical drug, a polypeptide drug, a protein drug and a gene drug that inhibits miR-327.

Further, the chemical drugs for inhibiting miR-327 generally refer to small molecule organic compounds. The protein drug for inhibiting miR-327 comprises natural protein and recombinant protein, including but not limited to antibody. The miR-327 inhibition "gene medicine" comprises but is not limited to nucleic acid fragments, such as DNA fragments and RNA fragments.

In one embodiment of the invention, the miR-327 inhibitor is an LNA inhibitor probe for antagonizing miR-327, which is customized by Exiqon corporation, the miR-327 inhibitor can target and inhibit miR-327, and the miR-327 inhibitor is complementary to the sequence of miR-327. Wherein, the nucleotide sequence of miR-327 comprises ACUUGAGGGGCAUGAGGAU.

Optionally, the FGF10 promoter comprises one or more of a chemical drug, a polypeptide drug, a protein drug, and a gene drug that promotes the increase of FGF 10.

Further, the chemical agents that promote the increase of FGF10 generally refer to small molecule organic compounds. The protein drugs promoting the increase of FGF10 comprise natural proteins and recombinant proteins, including but not limited to antibodies. The "gene-like drug" promoting the increase of FGF10 includes, but is not limited to, nucleic acid fragments, such as DNA fragments, RNA fragments. Specifically, the FGF10 promoter includes FGF10 protein and the like.

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

Optionally, the pharmaceutically acceptable carrier includes at least one of a solvent, a polymer, a liposome, a recombinant viral vector, and a eukaryotic recombinant expression vector, but is not limited thereto.

Wherein the solvent includes, but is not limited to, water, physiological saline, and other non-aqueous solvents.

The polymer includes one or more of polylysine, polyethyleneimine (branched and/or chain) and its modified substance, polyamidoamine dendrimer (PAMAM) and its derivative, polypropyleneimine dendrimer (PPI) and its derivative, chitosan, polylactic-co-glycolic acid (PLGA), polylactic acid, gelatin, cyclodextrin, sodium alginate, albumin, and hemoglobin, but is not limited thereto. Among them, polyethyleneimine and its modified products, PAMAM and its derivatives, PPI and its derivatives, chitosan, etc. may be referred to as cationic polymers.

The liposome can be prepared by self-assembly of cationic lipid, neutral auxiliary lipid, cholesterol, and phospholipid (such as soybean lecithin, yolk lecithin, cephalin, etc.), or can be prepared by inserting distearoylphosphatidylethanolamine-polyethylene glycol (DSPE-PEG) into phospholipid layer formed by phospholipid molecules.

The recombinant viral vector may include one or more of a lentiviral vector, an adenoviral vector, and a retroviral vector, but is not limited thereto.

In the present invention, the "pharmaceutically acceptable carrier" is used to transport the drug of the present invention to exert its intended effect. In general, delivery is from one organ or portion to another, and the carrier must be compatible with the pharmaceutical composition, not interfere with the biological activity of the drug, and be relatively non-toxic, e.g., the carrier enters the body without causing toxic side effects or having a severe reaction with the drug it carries, which does not adversely affect the patient.

The active pharmaceutical ingredients, namely the miR-327 inhibitor and/or the FGF10 promoter, can be dispersed or absorbed in the carrier to form a dispersion system, and can also be coated/encapsulated by the liposome, the polymer and the like to form a spherical structure (such as a nano capsule or a micro capsule). For example, when the miR-327 inhibitor is a nucleic acid fragment inhibiting miR-327, and the medicament for preventing and/or treating diabetes comprises but is not limited to a cationic polymer, a polypeptide, a protein medicament and the like which are wrapped, combined or blended with the nucleic acid fragment.

For another example, the miR-327 inhibitor encapsulated in the spherical structure can be subjected to sustained release, controlled release or targeted release, so that the drug can exert the optimal efficacy, and the stability of the drug can be improved. For example, albumin, gelatin, chitosan, polylactic acid can generally form microspheres that can disperse or encapsulate a drug.

Optionally, in the medicament for preventing and/or treating diabetes, the content of the miR-327 inhibitor is 5-20 mg/kg. Further preferably 5-10 mg/kg.

Optionally, in the medicament for preventing and/or treating diabetes, the content of the FGF10 promoter is 1-100 mg/kg.

Further, the content ratio of the miR-327 inhibitor to the FGF10 promoter is (0.2-20): 1.

in the medicine for preventing and/or treating diabetes, the miR-327 inhibitor can be selected as a single active ingredient, the FGF10 promoter can be selected as a single active ingredient, and the medicine can also contain the miR-327 inhibitor and the FGF10 promoter as active ingredients. This is also what is meant by "miR-327 inhibitor and/or FGF10 promoter" in the context of the present invention.

On the basis, the medicament for preventing and/or treating diabetes further comprises other pharmaceutically acceptable active ingredients. The miR-327 inhibitor and/or FGF10 promoter can act together with other pharmaceutically acceptable active ingredients to achieve the purpose of preventing and/or treating diabetes.

Wherein the other pharmaceutically acceptable active ingredients comprise one or more of glimepiride, glipizide, tolbutamide, troglitazone, voglibose and insulin.

Preferably, the medicament for preventing and/or treating diabetes contains one or two of a miR-327 inhibitor and an FGF10 promoter, and insulin; the miR-327 inhibitors and FGF10 boosters are also useful for increasing insulin sensitivity.

The term "insulin sensitivity" as used herein refers to the degree to which a body cell (or "subject") is sensitive to the action of insulin. The cell recognizes signals through an insulin receptor, and then transmits the signals through a PI3K/Akt/mTOR transduction pathway, so that the utilization of glucose by the cell is accelerated to reduce blood sugar. If the insulin sensitivity is low, also called insulin resistance (mainly referring to muscle cells), the body usually needs more endogenous or exogenous insulin to maintain blood sugar balance, and then hyperinsulinemia is generated, and the body is also susceptible to type II diabetes.

In the present invention, the other pharmaceutically acceptable active ingredient has an effect of preventing and/or treating diabetes. The Glimepiride (Glimepiride) is a second-generation sulfonylurea oral hypoglycemic agent, and the main mechanism of the hypoglycemic effect is to stimulate islet beta cells to secrete insulin, so that the sensitivity of peripheral tissues to the insulin is partially improved. The Glipizide (Glipizide) is a second-generation sulfonylurea oral hypoglycemic agent, can promote insulin secretion of islet beta cells, enhance the action of insulin on target tissues, stimulate islet alpha cells to inhibit glucagon secretion, and has the effects of inhibiting hepatic glycogenolysis, promoting muscle utilization and consuming glucose. The Tolbutamide (Tolbutamide) is a first-generation sulfonylurea oral hypoglycemic agent, selectively acts on islet beta cells, stimulates the secretion of insulin, and reduces blood sugar. The troglitazone (Noscal) is a thiazolidinedione drug, can improve insulin resistance, increase the sensitivity of human tissues to insulin, and enhance the action of insulin. The Voglibose (Voglibose) is an alpha glucosidase inhibitor, inhibits the activities of maltase, isomaltase, glucoamylase and sucrase, delays the digestion and absorption of sugar, and reduces the speed of decomposing starch into oligosaccharides such as maltose (disaccharide), maltotriose and dextrin (oligosaccharide) into glucose, and reduces the speed of decomposing sucrose into glucose and fructose, so that the absorption of glucose in intestinal tract is reduced, and the postprandial hyperglycemia is reduced. The Insulin (Insulin) is involved in regulating glucose metabolism, controlling blood sugar balance, and preventing and/or treating diabetes. Specifically, glimepiride, glipizide, tolbutamide, troglitazone, voglibose, insulin or other drugs for preventing and/or treating diabetes can be selected according to actual needs.

Optionally, the medicament for preventing and/or treating diabetes further comprises one or more of a diluent and an excipient.

The primary function of the diluent is to fill the weight or volume of the tablet to facilitate tableting. Optionally, the diluent comprises one or more of starches, sugars, celluloses, and inorganic salts. The excipient is additive except main active ingredients in the medicine, and comprises binder, filler, disintegrating agent, lubricant, wine, vinegar, medicinal juice, etc. in the tablet, matrix part in semi-solid preparation ointment and cream, antiseptic, antioxidant, corrective, aromatic, cosolvent, emulsifier, solubilizer, osmotic pressure regulator, colorant, etc. in the liquid preparation.

Alternatively, the form of the medicament for preventing and/or treating diabetes includes tablets, capsules, powders, granules, pills, syrups, solutions or suspensions. The form of the agent for the prophylaxis and/or treatment of diabetes depends on the actual use.

Alternatively, the agent for preventing and/or treating diabetes is administered orally or by injection.

Preferably, the agent for preventing and/or treating diabetes is administered by injection. In this case, the agent for preventing and/or treating diabetes is preferably a solution, for example, dissolved in water or physiological saline. Further, the injection is administered by intraperitoneal injection, subcutaneous injection, intramuscular injection or intravenous injection.

Alternatively, the agent for preventing and/or treating lipodystrophy may be administered locally or systemically.

Optionally, the agent for preventing and/or treating diabetes is administered in an amount of 5 to 20mg/kg body weight per day. Specifically, the amount of the agent for preventing and/or treating diabetes depends on various factors including, but not limited to, the desired biological activity and tolerance to the agent in the subject to be administered.

The invention provides application of a miR-327 inhibitor and/or FGF10 promoter in preparation of a medicament for preventing and/or treating diabetes. The miR-327 inhibitor and the FGF10 promoter can promote the increase of FGF10 in a fat matrix vascular component, promote the differentiation of fat cell precursors in the matrix vascular component to brown fat cells, promote the consumption of blood sugar and achieve the aim of preventing and/or treating diabetes. They also increase the body's sensitivity to insulin in the presence of insulin. The invention provides new applications of a miR-327 inhibitor and an FGF10 promoter. Opens up a new way for the treatment of diabetes, has more obvious prevention or treatment effect, and can effectively reduce the risk of people suffering from diabetes.

In a second aspect, the invention provides a medicament for preventing and/or treating diabetes, which comprises one or more of a miR-327 inhibitor and an FGF10 promoter, and a pharmaceutically acceptable carrier.

The miR-327 inhibitor is used for targeting miR-327 in Stromal Vascular Fraction (SVF) in adipose tissue, reduces expression of miR-327, promotes increase of fibroblast growth factor 10(FGF10) which is a target of miR-327 in non-adipose cells (namely various cells in the stromal vascular fraction), promotes differentiation of adipose cell precursors in the stromal vascular fraction to brown adipose cells, increases the body metabolic rate, and promotes blood sugar consumption. Thereby achieving the effect of preventing and/or treating diabetes.

Wherein the FGF10 promoter can promote the increase of FGF10 in fat-based Stromal Vascular Fraction (SVF), promote the differentiation of fat cell precursors in SVF to brown fat cells, increase the metabolic rate of the body, and promote the consumption of blood sugar. Can also be used for preventing and/or treating diabetes.

Wherein the pharmaceutically acceptable carrier is as described in the first aspect of the invention.

Wherein the medicament for preventing and/or treating diabetes also comprises insulin. At this time, both the miR-327 inhibitor and the FGF10 promoter can improve the sensitivity of the body to insulin in the presence of insulin.

The second aspect of the invention provides a medicament for preventing and/or treating diabetes, which comprises one or more of a miR-327 inhibitor and an FGF10 promoter, and a pharmaceutically acceptable carrier. The medicine can promote the increase of FGF10 in blood vessel components of fat matrix, promote the differentiation of fat cell precursors in the blood vessel components of matrix to brown fat cells, increase the metabolic rate of organism, promote the consumption of blood sugar, and further improve diabetes.

Advantages of embodiments of the invention will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of embodiments of the invention.

Drawings

FIG. 1 shows the histological examination structure of control group 2 and control group 3 which were not or not stimulated with the drug. (ii) a

FIG. 2 is a graph quantifying the mean diameter of adipocytes in control group 2 and control group 3 that were not subjected to cold stimuli;

FIG. 3 is a graph showing the results of testing glucose tolerance (a), HOMA-IR (b), and core body temperature (c) of control group 2 and control group 3, which were given cold stimuli or not;

FIG. 4 shows the results of histological examination of the test group and the control group 1, which were not or not given cold stimuli;

FIG. 5 is a test result of the maximum oxygen uptake capacity of the experimental group and the control group 1, to which cold stimuli were given or not;

FIG. 6 shows the results of the test of the insulin resistance steady state model evaluation (HOMA-IR) index of the experimental group and the control group 1, which were administered with cold stimulation or not;

fig. 7 is a test result of core body temperature of the experimental group and the control group 1, to which cold stimuli were given or not.

Detailed Description

While the following is a description of the preferred embodiments of the present invention, it should be noted that those skilled in the art can make various modifications and improvements without departing from the principle of the embodiments of the present invention, and such modifications and improvements are considered to be within the scope of the embodiments of the present invention.

Experimental groups: a customized miR-327 antagonism LNA inhibitor probe is selected as a miR-327 inhibitor (the miR-327 inhibitor can target and inhibit miR-327, the sequence of the miR-327 inhibitor is complementary with the nucleotide sequence of miR-327, wherein the nucleotide sequence of miR-327 comprises ACUUGAGGGGCAUGAGGAU). 12 female C57BL/6 mice, 8 weeks old, were placed under thermoneutral conditions (30 ℃) and the miR-327 inhibitors described above were injected subcutaneously into the mice at a dose of 8mg/kg 1 time per week, followed by cold (4 ℃) or drug stimulation (e.g.beta adrenoceptor agonists such as CL316243) for a total of 2 weeks. After 2 weeks the following experiments were performed:

(1)2 weeks later 6 of these mice were treated with excess CO₂Sacrificed and the subcutaneous fat was dissected carefully, the weight was measured on a precision balance and statistically analyzed to obtain the mouse adipose tissue weighing result. Meanwhile, the histological detection of the adipose tissues of the mouse specifically comprises the following steps: adipose tissue was fixed using 4% PFA, sliced, digested with cathepsin K (20mM), and permeabilized with 100% methanol. The samples were blocked overnight with 3% milk in 0.1% Triton X-100PBS and incubated with a primary antibody such as anti-CD 31(1:100, Cat. AF3628, R)&D) And anti-Perilipin (1:200, catalog No. 20R-PP004, Fitzgerald Industries), anti-UCP 1(1:200, Abcam), and the like. After washing with PBS, the samples were blocked with 3% milk and incubated with a fluorescently labeled secondary antibody (1: 300, Thermo Fisher Scientific Inc.) for 2 hours at room temperature. After washing, the samples were mounted, observed and measured on a Confocal microscope (LSM510Confocal, Zeiss).

(2) The core body temperature of 6 of the mice was measured after 2 weeks as follows: the body temperature of each group of mice was measured using a MicroTherma 2T thermometer with the probe inserted into the rectum of the mice (cat # TW2 and RET-3, Agnto's AB).

These female mice were then subjected to an oral glucose tolerance test, which specifically included: fasting was for 6 hours, during which time water was allowed to drink freely. Glucose catalog number G8270, Sigma-Aldrich) was then dissolved in physiological saline and the mice were fed at a dose of 15mg/G mouse body weight. Tail vein blood was collected at each time point (0, 15, 30, 60, 120 minutes) and blood glucose levels were measured using an Accu-Chek Aviva glucometer and dipstick (Roche Diagnostics).

On day 2, 6 female mice were subjected to insulin resistance homeostasis model evaluation (HOMA-IR) index experiments. Among them, HOMA-IR is a well-established method for detecting insulin resistance. Fasting insulin levels in mouse serum samples were detected using an insulin ELISA kit. Fasting blood glucose levels were measured using an Aviva glucometer. The HOMA-IR index was calculated using the following formula: fasting plasma glucose (mg dL-1). times.fasting insulin (mU L-1)/405.

On day 3, the 6 female mice were subjected to metabolic level determination, comprising the following steps: oxygen consumption was used to quantify metabolic rate and Norepinephrine (NE) -induced thermogenesis. The Oxymax CLAMS-HC system (Columbus Instruments) was used. The oxygen sensor was heated for at least 6 hours and calibrated after stabilization using two reference gases. The two gases are 100% nitrogen and 20.5% O₂Or 0.5% CO₂And (4) mixing the gases. To measure basal metabolism, mice were transferred to an Oxymax chamber and basal metabolism was measured under different conditions. The chamber was allowed to settle for 24 hours and the measurement was carried out for 48-72 hours before the measurement. The Oxymax system was maintained at 30 ℃ of heat neutrality while measuring norepinephrine-induced thermogenesis. Norepinephrine was injected subcutaneously at a dose of 1mg/kg body weight, based on the metabolic rate 24 hours prior to norepinephrine injection. The area under the curve of oxygen consumption between 30 minutes before injection and 140 minutes after injection was calculated and compared between groups.

Control group 1: no-effect miR inhibitor drugs (random sequences) that do not inhibit miR-327 at a dose of 8mg/kg were injected subcutaneously weekly into white fat of 12C 57BL/6 mice of 8 weeks of age, then given cold (4 ℃) stimulation, injected 1 time per week for a total of 2 weeks; 2 weeks later 6 of these mice were given excess CO₂Sacrifice, weighing its adipose tissue and performing histological examination; the other 6 mice were subjected to core body temperature measurement, oral glucose tolerance test, HOMA-IR index test, and metabolic level measurement in this order.

Control group 2: lentiviral vectors (Vector Biosystems) carrying non-functional miR were used at 1X 10⁹PFU/dose per dose was injected subcutaneously into white fat of 12C 57BL/6 mice 8 weeks old, followed by cold (4 ℃) stimulation one week later, 1 injection per week for a total of 2 weeks; 2 weeks later 6 of these mice were given excess CO₂Sacrifice, weighing its adipose tissue and performing histological examination; the other 6 mice were subjected to core body temperature measurement and oral glucose tolerance in sequenceQuantitative test, HOMA-IR index test and metabolic level determination.

Control group 3: adenovirus overexpressing mmu-miR-327 (Ad-EF1 a-mmu-miR-327-eGFP; 1X 109 PFU; Vector Biosystems) at 1X 10⁹PFU/dose per dose was injected subcutaneously into white fat of 12C 57BL/6 mice 8 weeks old weekly, followed by cold (4 ℃) stimulation one week later, 1 injection weekly for a total of 2 weeks; (ii) a 2 weeks later 6 of these mice were given excess CO₂Sacrifice, weighing its adipose tissue and performing histological examination; the other 6 mice were subjected to core body temperature measurement, oral glucose tolerance test, HOMA-IR index test, and metabolic level measurement in this order.

The histological results of the control group 2 and the control group 3 are shown in fig. 1 and 2, and fig. 1 is the histological result of the control group 2 and the control group 3 when the drug is not stimulated. Fig. 2 is a graph showing the quantification of the mean diameter of adipocytes in the absence and presence of cold stimuli given to control group 2 and control group 3. Wherein Ad-miRNC represents a control group 2, and Ad-miR-327 represents a control group 3. In fig. 1, H & E represents morphological staining of adipose tissue using hematoxylin and eosin. UCP1 staining showed brownish locations, PERI showed adipocyte morphology, DAPI counterstained the nuclei.

As can be seen from FIGS. 1 and 2, in the control group 3, after injecting an adenovirus expressing miR-327 in a mouse and expressing miR-327 in its adipose tissue (which may be abbreviated as Ad-miR-327), the size of adipocytes was unchanged compared to the control group 2 without stimulation to promote browning by cold, drugs, etc. (wherein double arrows indicate the size of adipocytes). In the presence of cold, drugs and other stimuli promoting browning, the cell diameter of miR-327-expressing adipose tissues is not reduced (i.e., slightly increased), UCP1 staining shows that the browning degree is reduced, and arrows indicate sites with obvious browning.

FIG. 3 is a graph showing the results of testing the glucose tolerance (a), HOMA-IR (b), and core body temperature (c) of control group 2 and control group 3, which were not or not given cold stimuli. FIGS. 3 (a) and (b) show that after expression of miR-327 in adipose tissue, neither cold nor cold stimulation affected insulin sensitivity. Where ns represents no significant difference. Fig. 3 (c) shows that, in the absence of cold, drug, or other stimulus that promotes browning, the body core temperature of control group 3 was unchanged from control group 2 after expression of miR-327 in adipose tissue. In the presence of cold, drugs and other stimuli that promote browning, the body core temperature of mice expressing miR-327 in their fat decreases.

The results of fig. 1 to 3 demonstrate that introduction of miR-327 into mouse adipose tissue using a viral vector significantly inhibits browning of white adipocytes, hindering the lipid burning effect. Reduce the energy metabolism of the mice.

The results of histological detection and metabolic test of the experimental group and the control group 1 are shown in fig. 4-7, wherein Inh-mircn represents the control group 1, and Inh-miR-327 represents the experimental group.

As can be seen from fig. 4, after the mice were injected with the miR-327 inhibitor, the size of adipocytes (double arrows indicate the size of adipocytes) had been significantly reduced compared to control 1 in the absence of the stimulus for promoting browning, such as cold, drugs, etc., and the staining of UCP1 showed that the degree of browning was significantly increased. In the presence of cold, drugs and other stimuli promoting browning, mice injected with the miR-327 inhibitor further decrease in adipocyte size, and staining by UCP1 shows that the degree of browning is further increased, with arrows indicating sites of marked browning.

As seen from FIG. 5, the maximum oxygen uptake capacity (VO) as a metabolic index of miR-327 inhibitor injected into mice is obtained without stimulation of cold, drugs, etc. for promoting browning₂) Significantly improved compared to control 1. If the mice are stimulated by cold, medicaments and the like to promote browning, the metabolic indexes of the mice injected with the miR-327 inhibitor are further improved.

As seen in FIG. 6, the HOMA-IR has been significantly reduced in non-cold conditions after the mice are injected with the miR-327 inhibitor, indicating that the miR-327 inhibitor can increase insulin sensitivity at room temperature.

As seen in FIG. 7, after injection of the miR-327 inhibitor, the body core temperature of the mice can be increased and the metabolic rate can be increased already under non-cold conditions.

The results of fig. 4-7 demonstrate that the use of the miR-327 inhibitor can significantly promote browning of white fat in mice, increase the metabolic rate of the body, promote blood glucose consumption, and increase insulin sensitivity.

The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the present invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.

Claims

The application of the miR-327 inhibitor or the combination of the miR-327 inhibitor and the FGF10 promoter in preparing a medicament for treating diabetes.
2. The use of claim 1, wherein the miR-327 inhibitor and the FGF10 promoter both promote an increase in FGF10 in the stromal vascular fraction of adipose tissue, promote differentiation of adipocyte precursors to brown adipocytes in the stromal vascular fraction, increase the rate of body metabolism, promote blood glucose consumption, and increase insulin sensitivity; wherein the miR-327 inhibitor promotes the increase of FGF10 as a target of miR-327 by inhibiting expression of miR-327 in the vascular component of fat stroma.
3. The use of claim 1, wherein said miR-327 inhibitor comprises one or more of a chemical, polypeptide, protein, and gene drug that inhibits miR-327;

the FGF10 promoter includes one or more of a chemical drug, a polypeptide drug, a protein drug, and a gene drug that promotes the increase of FGF 10.
4. The use of claim 1, wherein the medicament for the treatment of diabetes comprises both a miR-327 inhibitor and a FGF10 promoter.
5. The use of claim 1, wherein the medicament for the treatment of diabetes further comprises a pharmaceutically acceptable carrier; the pharmaceutically acceptable carrier comprises at least one of a solvent, a polymer, a liposome, a recombinant viral vector and a eukaryotic recombinant expression vector.
6. The use according to claim 1, wherein the medicament for the treatment of diabetes further comprises other pharmaceutically acceptable active ingredients; the other pharmaceutically acceptable active ingredients include one or more of glimepiride, glipizide, tolbutamide, troglitazone, voglibose and insulin.
7. The use of claim 1, wherein the diabetes treatment medicament comprises a miR-327 inhibitor or a miR-327 inhibitor and FGF10 promoter, and the diabetes treatment medicament further comprises insulin.
8. The medicine for treating diabetes is characterized by comprising a miR-327 inhibitor, an FGF10 promoter and a pharmaceutically acceptable carrier.
9. The medicament for treating diabetes mellitus of claim 8, wherein the content of the miR-327 inhibitor in the medicament for treating diabetes mellitus is 5-20 mg/kg.
10. The medicament for treating diabetes according to claim 8, further comprising insulin; the miR-327 inhibitors and FGF10 boosters are also useful for increasing insulin sensitivity.