Application of hydrogel-loaded long-acting insulin receptor antagonist in establishment of gestational diabetes animal model
Through the hydrogel-loaded long-acting insulin receptor antagonist, subcutaneous injection every 2 days, a stable and reliable model of gestational diabetes mice was established, solving the toxicity, time and time in the existing model establishment method. The low modeling rate and other problems have achieved a simulation similar to the pathogenesis of gestational diabetes in humans.
Patent Information
- Application Number
- CN202411651957.2
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-11-19
- Publication Date
- 2025-02-25
AI Technical Summary
The existing methods for establishing animal models of gestational diabetes have problems such as the toxicity of streptozotocin, high sugar and high fat induction methods that require long-term feeding and low modeling rate, short biological half-life of insulin receptor antagonists and poor compliance, which is difficult to Effectively simulate the pathophysiological status of gestational diabetes.
Using a hydrogel-loaded long-acting insulin receptor antagonist, gestational diabetes was established by subcutaneous injection of 60 nmol/kg of phenylboronic acid-hyaluronic acid hydrogel-loaded insulin receptor antagonist every 2 days starting from 7 days of pregnancy in animals. Animal model.
A stable and reliable mouse model similar to the pathogenesis of human gestational diabetes was successfully established. The modeling time was short and the success rate was high, and there was no obvious toxic effect on the female mice and embryos, which was consistent with the characteristics of human gestational diabetes.
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Figure CN119499165A_ABST
Abstract
Description
Technical Field
[0001] The invention relates to the technical field of diabetes research, and in particular to a method for using a hydrogel-loaded long-acting insulin receptor antagonist in establishing an animal model of gestational diabetes. Background Art
[0002] Gestational diabetes is a phenomenon of hyperglycemia caused by relatively insufficient insulin secretion or insulin resistance in the middle and late stages of pregnancy due to physiological changes during pregnancy. It is mainly manifested in decreased insulin sensitivity during pregnancy and increased blood sugar caused by insufficient insulin utilization. The blood sugar of patients with gestational diabetes can mostly return to normal after the placenta is delivered, but gestational diabetes can induce adverse pregnancy outcomes and have a negative impact on the long-term health of mothers and offspring, increase the risk of obesity and type 2 diabetes in offspring in adulthood, and can also cause neurodevelopmental damage in offspring.
[0003] Due to the lack of an animal model that effectively simulates the pathophysiological state of gestational diabetes in pregnant women, the development of drugs for preventing and treating gestational diabetes and its impact on the health of offspring has been slow. Therefore, the establishment of a reliable and easy-to-operate animal model of gestational diabetes will greatly promote the development of related prevention and treatment methods and drugs, including the screening and evaluation of protective drugs. Although methods for inducing gestational diabetes models have been reported, including induction by streptozotocin, natural induction by long-term high-sugar and high-fat diet, or induction by insulin receptor antagonists. At present, these methods for establishing gestational diabetes still have some shortcomings. For example, streptozotocin injection mainly simulates the type I diabetes model rather than the typical model of relatively insufficient insulin secretion in gestational diabetes. In addition, streptozotocin is toxic and can cause death of experimental animals, and is embryotoxic, so it is not suitable for studying the effects of gestational diabetes on the health of offspring. The high-sugar and high-fat induced obesity-induced gestational diabetes model requires a long period of high-sugar and high-fat diet feeding, and the modeling rate is low, with many reports showing about 50%. Moreover, this method simulates the diet-induced obesity-induced gestational diabetes model, but cannot simulate the situation of normal weight and simple insulin resistance. Insulin receptor antagonists are polypeptides composed of 43 amino acids. After entering the animal body, their biological half-life is short, about 2 to 3 hours. They need to be administered daily during modeling. Animal compliance is poor, and blood sugar is maintained for a short time, affecting the experimental results. Summary of the invention
[0004] The present invention provides a method for using a hydrogel-loaded long-acting insulin receptor antagonist in establishing an animal model of gestational diabetes, which has wide application value in the research of the influence of maternal gestational diabetes on the health of offspring and protective drugs.
[0005] The objective of the present invention is achieved through the following technical solutions:
[0006] The invention discloses an application of a hydrogel-loaded long-acting insulin receptor antagonist in establishing an animal model of gestational diabetes. Starting from the 7th day of pregnancy, 60 nmol / kg of insulin receptor antagonist loaded with phenylboronic acid-hyaluronic acid hydrogel is subcutaneously injected every 2 days to establish an animal model of gestational diabetes.
[0007] Furthermore, the animal is a C57BL / 6J female mouse.
[0008] Furthermore, the method specifically comprises:
[0009] 1) Female mice were grouped and fed: 11-week-old C57BL / 6J female mice were randomly divided into two groups after one week of adaptive feeding, namely, gestational diabetes group (GDM) and control group (CON);
[0010] 2) C57BL / 6J female mice were pregnant and the pregnancy status was observed;
[0011] 3) preparing phenylboronic acid-hyaluronic acid hydrogel injection and phenylboronic acid-hyaluronic acid hydrogel loaded insulin receptor antagonist;
[0012] 4) Insulin receptor antagonist loaded with phenylboronic acid-hyaluronic acid hydrogel induced gestational diabetes: Starting from the 7th day of pregnancy of C57BL / 6J female mice, the GDM group was subcutaneously injected with 60nmol / kg of insulin receptor antagonist loaded with phenylboronic acid-hyaluronic acid hydrogel every 2 days to establish an animal model of gestational diabetes; the CON group was subcutaneously injected with phenylboronic acid-hyaluronic acid hydrogel injection every 2 days as a control.
[0013] 5) Seven days after administration, pregnant mice were given an oral glucose tolerance test.
[0014] Furthermore, the step 1) specifically includes: after 11-week-old C57BL / 6J female mice were adaptively fed with a basic feed for 1 week, they were randomly divided into 2 groups with the same number of mice in each group, and the basic fasting blood glucose was measured.
[0015] Furthermore, the step 2) is specifically as follows: C57BL / 6J female mice are mated with male mice in a 2:1 ratio, and the vaginal plug is observed. The observation of the vaginal plug is recorded as pregnancy day 0.
[0016] Furthermore, the step 3) is specifically as follows: preparing an injectable phenylboronic acid-hyaluronic acid hydrogel injection solution with phenylboronic acid-hyaluronic acid hydrogel and PBS buffer in a mass: volume ratio of 1:4 (w:v, 1:4)
[0017] Furthermore, in the step 3), 1 mg of insulin receptor antagonist is added to 1 ml of PBS to dissolve to 206 nmol / mL, diluted 30 times with PBS buffer, and then mixed with phenylboronic acid-hyaluronic acid hydrogel in a ratio of 4:1, vortexed at room temperature, and ultrasonicated for 10 minutes to allow the phenylboronic acid-hyaluronic acid hydrogel to fully load the insulin receptor antagonist.
[0018] Furthermore, the step 4) is specifically as follows: when the C57BL / 6J female mice are 7 days pregnant, the fasting blood glucose of the pregnant mice is measured on the 7th day of pregnancy; starting from the 7th day of pregnancy of the C57BL / 6J female mice, according to the weight of the animals, the GDM group is subcutaneously injected with 60 nmol / kg of insulin receptor antagonist loaded with phenylboronic acid-hyaluronic acid every 2 days, with a solution concentration of 6.7 nmol / ml and a total injection volume of <150 μl, to establish an animal model of gestational diabetes.
[0019] Furthermore, in step 4), the CON group was subcutaneously injected with phenylboric acid-hyaluronan injection every 2 days starting from the 7th day of pregnancy as a control.
[0020] Furthermore, in the step 4), the administration time of the phenylboronic acid-hyaluronic acid loaded insulin receptor antagonist and the phenylboronic acid-hyaluronic acid injection is 5 pm every day until the day before delivery.
[0021] Furthermore, the step 5) is specifically as follows: after 7 days of administration, the GDM and CON animals were fasted for 10 hours, blood was collected from the tail vein to determine the fasting blood glucose, and then 1.2 g / kg glucose solution was gavaged, and blood glucose was measured 30 minutes, 60 minutes, 120 minutes, and 180 minutes after gavage, and an oral glucose tolerance test was performed to determine the animal modeling status.
[0022] The present invention has the following beneficial effects:
[0023] Different from the previous gestational diabetes mouse or rat models with long modeling time, low success rate, high animal mortality rate, and embryotoxicity, the present invention has established a stable and reliable mouse model similar to the human pathogenesis, with short modeling time, high success rate, and no obvious toxic effects on mother mice and embryos, which is consistent with the characteristics of human gestational diabetes. BRIEF DESCRIPTION OF THE DRAWINGS
[0024] Figure 1 is the initial baseline fasting blood glucose of C57BL / 6J mice in the embodiment of the present invention;
[0025] Figure 2 is the fasting blood sugar of C57BL / 6J mice on the 7th day of pregnancy in the embodiment of the present invention;
[0026] Figure 3This is an oral glucose tolerance test in C57BL / 6J mice after 7 days of administration in the examples of the present invention. DETAILED DESCRIPTION
[0027] The present invention is further described in detail below by means of specific embodiments in combination with the accompanying drawings. It should be understood that the embodiments are only exemplary and cannot be construed as limiting the scope of protection of the present invention.
[0028] A hydrogel-loaded long-acting insulin receptor antagonist is used in the establishment of an animal model of gestational diabetes. Starting from the 7th day of pregnancy, 60nmol / kg of phenylboronic acid-hyaluronic acid-loaded insulin receptor antagonist is subcutaneously injected every 2 days to establish an animal model of gestational diabetes.
[0029] Furthermore, the animal is a C57BL / 6J female mouse.
[0030] Furthermore, the method specifically comprises:
[0031] 1) Female mice were grouped and fed: 11-week-old C57BL / 6J female mice were randomly divided into two groups after one week of adaptive feeding, namely, gestational diabetes group (GDM) and control group (CON);
[0032] 2) C57BL / 6J female mice were pregnant and the pregnancy status was observed;
[0033] 3) preparing phenylboronic acid-hyaluronic acid hydrogel injection and phenylboronic acid-hyaluronic acid hydrogel loaded insulin receptor antagonist;
[0034] 4) Insulin receptor antagonist loaded with phenylboronic acid-hyaluronic acid hydrogel induced gestational diabetes: Starting from the 7th day of pregnancy of C57BL / 6J female mice, the GDM group was subcutaneously injected with 60nmol / kg of insulin receptor antagonist loaded with phenylboronic acid-hyaluronic acid hydrogel every 2 days to establish an animal model of gestational diabetes; the CON group was subcutaneously injected with phenylboronic acid-hyaluronic acid hydrogel injection every 2 days as a control.
[0035] 5) Seven days after administration, pregnant mice were given an oral glucose tolerance test.
[0036] Furthermore, the step 1) specifically includes: after 11-week-old C57BL / 6J female mice were adaptively fed with a basic feed for 1 week, they were randomly divided into 2 groups with the same number of mice in each group, and the basic fasting blood glucose was measured.
[0037] Furthermore, the step 2) is specifically as follows: C57BL / 6J female mice are mated with male mice in a 2:1 ratio, and the vaginal plug is observed. The observation of the vaginal plug is recorded as pregnancy day 0.
[0038] Furthermore, the step 3) is specifically as follows: preparing an injectable phenylboronic acid-hyaluronic acid hydrogel injection solution with phenylboronic acid-hyaluronic acid hydrogel and PBS buffer in a mass: volume ratio of 1:4 (w:v, 1:4)
[0039] Furthermore, in the step 3), 1 mg of insulin receptor antagonist is added to 1 ml of PBS to dissolve to 206 nmol / mL, diluted 30 times with PBS buffer, and then mixed with phenylboronic acid-hyaluronic acid hydrogel in a ratio of 4:1, vortexed at room temperature, and ultrasonicated for 10 minutes to allow the phenylboronic acid-hyaluronic acid hydrogel to fully load the insulin receptor antagonist.
[0040] Furthermore, the step 4) is specifically as follows: when the C57BL / 6J female mice are 7 days pregnant, the fasting blood glucose of the pregnant mice is measured on the 7th day of pregnancy; starting from the 7th day of pregnancy of the C57BL / 6J female mice, according to the weight of the animals, the GDM group is subcutaneously injected with 60 nmol / kg of insulin receptor antagonist loaded with phenylboronic acid-hyaluronic acid every 2 days, with a solution concentration of 6.7 nmol / ml and a total injection volume of <150 μl, to establish an animal model of gestational diabetes.
[0041] Furthermore, in step 4), the CON group was subcutaneously injected with phenylboric acid-hyaluronan injection every 2 days starting from the 7th day of pregnancy as a control.
[0042] Furthermore, in the step 4), the administration time of the phenylboronic acid-hyaluronic acid loaded insulin receptor antagonist and the phenylboronic acid-hyaluronic acid injection is 17:00 in the afternoon and the administration is continued until delivery.
[0043] Furthermore, the step 5) is specifically as follows: after 7 days of administration, the GDM and CON animals were fasted for 10 hours, blood was collected from the tail vein to determine the fasting blood glucose, and then 1.2 g / kg glucose solution was gavaged, and blood glucose was measured 30 minutes, 60 minutes, 120 minutes, and 180 minutes after gavage, and an oral glucose tolerance test was performed to determine the animal modeling status.
[0044] Example
[0045] This embodiment relates to the use of a hydrogel-loaded long-acting insulin receptor antagonist in establishing an animal model of gestational diabetes, including:
[0046] 1 Experimental Materials
[0047] Twenty SPF-grade 11-week-old C57BL / 6J female mice were purchased from Henan Sikebes Biotechnology Co., Ltd. The experimental animals were kept in an SPF-grade laboratory with an ambient temperature of 24±2℃, a humidity of 40-70%, and a light-dark cycle of 1:1. They were randomly divided into 4 groups, with 10 mice in each group. Sterile basic feed containing 10% fat was purchased from Beijing Huafukang Experimental Animal Co., Ltd.
[0048] 2 Main reagents
[0049] Insulin receptor antagonist: purchased from Nanjing Jiepeptide Biotechnology Co., Ltd.
[0050] Phenylboric acid-hyaluronic acid hydrogel: purchased from Shenzhen Meiluo Technology Co., Ltd.
[0051] 3. Preparation of main solutions
[0052] PBS. 6.7nmol / ml phenylboronic acid-hyaluronic acid loaded insulin receptor antagonist: Weigh 1mg insulin receptor antagonist powder, dissolve in 1ml PBS to prepare 200nmol / mL stock solution, dilute the stock solution 30 times with PBS buffer, add it to the phenylboronic acid-hyaluronic acid hydrogel in the ratio of insulin receptor antagonist solution volume: mass (4:1), vortex and sonicate to fully load the phenylboronic acid-hyaluronic acid hydrogel with insulin receptor antagonist; calculate the required dose based on 60nmol / kg animal body weight, and administer once every 2 days by subcutaneous injection.
[0053] 4 Main instruments
[0054] Blood glucose meter: Abbott blood glucose tester, Abbott blood glucose test strips.
[0055] 5 Specific experimental methods
[0056] 5.1Basal blood glucose in C57BL / 6J female mice
[0057] (1) After 11-week-old female mice were adaptively fed with a 10% fat-containing basal diet for 1 week, they were randomly divided into 2 groups, 10 mice in each group. The animals were fasted for 10-12 hours but were not deprived of water.
[0058] (2) Blood was collected from the tail vein and baseline fasting blood glucose was measured using an Abbott blood glucose meter and matching blood glucose test strips.
[0059] 5.2 Pregnancy of C57BL / 6J female mice
[0060] (1) C57BL / 6J female mice were mated with male mice in a 2:1 ratio. The vaginal plug was observed at 6:00 a.m. and 20:00 p.m. The day of pregnancy was recorded as 0 when the vaginal plug was observed.
[0061] (2) Weigh the mice daily and observe their weight gain during pregnancy.
[0062] 5.3 Fasting blood sugar on the 7th day of pregnancy
[0063] (1) On 7 days of gestation, mice were fasted for 10 hours and their fasting blood glucose was measured by blood sampling from the tail vein.
[0064] 5.4 Establishment of gestational diabetes model
[0065] (1) For C57BL / 6J female mice in the GDM group, on 7 days of pregnancy, 60 nmol / kg of insulin receptor antagonist loaded with phenylboronic acid-hyaluronic acid was subcutaneously injected according to the animal weight. The solution concentration was 6.7 nmol / ml, and the total injection volume was less than 150 ul, which was easy for the animals to tolerate.
[0066] (2) The injection is given every 2 days, at approximately 17:00 in the afternoon.
[0067] 5.5 Oral Glucose Tolerance Test
[0068] (1) Seven days after administration, the animals were subjected to an oral glucose tolerance test on gestational day 14. Glucose solution was administered by gavage at 1.2 g / kg.
[0069] (2) After the pregnant mice were fasted for 10 hours, blood was collected from the tail vein to measure fasting blood glucose. The mice were then given glucose solution by gavage, and blood glucose was measured 30 minutes, 60 minutes, 120 minutes, and 180 minutes after gavage, all using the tail vein blood sampling method.
[0070] 5.6 Birth weight and lactation weight of offspring
[0071] (1) The mother mice gave birth naturally and the birth weight of the pups was measured. The mother mice were allowed free access to water and food.
[0072] (2) The offspring were breastfed for 4 weeks and their body weights were measured every week.
[0073] The following are the experimental results of this embodiment:
[0074] The method of subcutaneous injection of 60 nmol / kg of insulin receptor antagonist loaded with phenylboronic acid-hyaluronic acid to 7-day pregnant mice every 2 days until delivery can successfully induce an animal model of gestational diabetes characterized by insulin resistance. Mice develop impaired glucose tolerance in the middle and late stages of pregnancy.
[0075] The successful modeling criteria include (1) abnormal glucose tolerance in late pregnancy of maternal rats; (2) the offspring are prone to acquired obesity after birth. The modeling model in this experiment meets the above modeling criteria, with a modeling success rate of 100%, and no obvious side effects or complications.
[0076] The body weight of the female mice before and after one week of adaptive feeding is shown in Table 1:
[0077] Table 1 Basal body weight of female mice
[0078]
[0079] Note: Data are expressed as mean ± standard deviation; basic control group (CON), gestational diabetes group (GDM).
[0080] Table 1 shows that there was no difference in the initial body weight after the animals were randomly divided into two groups and in the body weight after 1 week of adaptive feeding.
[0081] like Figure 1 As shown, there was no difference in baseline blood glucose between the two groups in female mice before pregnancy.
[0082] like Figure 2 As shown, there was no difference in fasting blood glucose between the two groups in female mice at 7 days of pregnancy.
[0083] like Figure 3 As shown, 7 days after administration (gestational age 14), the insulin receptor antagonist loaded with phenylboronic acid-hyaluronic acid caused abnormal glucose tolerance in mice, while the phenylboronic acid-hyaluronic acid hydrogel injection did not cause abnormal glucose tolerance in animals.
[0084] The effects of phenylboronic acid-hyaluronic acid loaded insulin receptor antagonist on the growth and development of offspring mice at birth and during lactation are shown in Table 2:
[0085] Table 2 Effects of maternal GDM induced by phenylboronic acid-hyaluronic acid loaded insulin receptor antagonist on the growth and development of offspring
[0086]
[0087] Note: Basic control group (CON), gestational diabetes group (GDM); * Compared with CON group, P<0.05
[0088] Table 2 shows that maternal GDM induced by phenylboronic acid-hyaluronan loaded insulin receptor antagonist can cause an increase in birth weight of mice. Starting from the fourth week after birth, the weight of the offspring in the GDM group was higher than that in the CON group. The offspring in the GDM group had a higher susceptibility to obesity, and the weight gain was significantly higher than that in the control group.
[0089] The gestational diabetes model was induced by subcutaneous injection of 60 nmol / kg phenylboronic acid-hyaluronan-loaded insulin receptor antagonist every 2 days starting from 7 days of pregnancy, without causing animal mortality, complications or side effects.
[0090] Gestational diabetic animals induced by subcutaneous injection of 60 nmol / kg phenylboronic acid-hyaluronan-loaded insulin receptor antagonist every 2 days starting from gestational day 7 developed glucose tolerance abnormalities at gestational day 14, affecting the obesity susceptibility of offspring during the weaning period.
[0091] The experimental results showed that the gestational diabetes model induced by subcutaneous injection of 60nmol / kg phenylboronic acid-hyaluronic acid-loaded insulin receptor antagonist every 2 days starting from the 7th day of pregnancy caused insulin resistance, insufficient insulin utilization, impaired glucose tolerance, and clinical manifestations of gestational diabetes in pregnant mice, and a gestational diabetes mouse animal model was successfully established.
[0092] The modeling method of the present invention can form a stable and standardized gestational diabetes model, which can be used for the screening of gestational diabetes drugs and the study of the effect of gestational diabetes on the growth and development of offspring. Compared with other drug-induced gestational diabetes models, the insulin resistance induced by the insulin receptor antagonist loaded with phenylboronic acid-hyaluronic acid better simulates the human pathogenesis, and is more long-lasting and has higher animal compliance than the simple insulin receptor antagonist induction, and has greater application value in the screening of gestational diabetes treatment drugs and the study of the effect of gestational diabetes on the growth and development of offspring.
[0093] The above contents are further detailed descriptions of the present invention in combination with specific implementation methods, and it cannot be determined that the specific implementation of the present invention is limited to these descriptions. For ordinary technicians in the technical field of the present invention, several simple deductions or substitutions can be made without departing from the concept of the present invention, which should be regarded as falling within the scope of protection of the present invention.
Claims
1. Use of a hydrogel-loaded long-acting insulin receptor antagonist in establishing an animal model of gestational diabetes, characterized in that: Starting from the 7th day of pregnancy, 60 nmol / kg of insulin receptor antagonist peptide loaded with phenylboronic acid-hyaluronic acid hydrogel was subcutaneously injected every 2 days to establish an animal model of gestational diabetes.
2. The method for using a hydrogel-loaded long-acting insulin receptor antagonist in establishing an animal model of gestational diabetes mellitus as claimed in claim 1, characterized in that: The animals were C57BL / 6J female mice.
3. Use of a hydrogel-loaded long-acting insulin receptor antagonist as claimed in claim 2 in establishing an animal model of gestational diabetes, characterized in that: The method for establishing an animal model of gestational diabetes mellitus comprises the following steps: 1) Female mice were grouped and fed: 11-week-old C57BL / 6J female mice were randomly divided into two groups after one week of adaptive feeding, namely, gestational diabetes group (GDM) and control group (CON); 2) C57BL / 6J female mice were pregnant and the pregnancy status was observed; 3) preparing phenylboronic acid-hyaluronic acid hydrogel injection and phenylboronic acid-hyaluronic acid hydrogel loaded insulin receptor antagonist; 4) Insulin receptor antagonist loaded with phenylboronic acid-hyaluronic acid hydrogel induced gestational diabetes: Starting from the 7th day of pregnancy of C57BL / 6J female mice, the GDM group was subcutaneously injected with 60nmol / kg of insulin receptor antagonist loaded with phenylboronic acid-hyaluronic acid hydrogel every 2 days to establish an animal model of gestational diabetes; the CON group was subcutaneously injected with phenylboronic acid-hyaluronic acid hydrogel injection every 2 days as a control; 5) Seven days after administration, pregnant mice were given an oral glucose tolerance test.
4. Use of a hydrogel-loaded long-acting insulin receptor antagonist in establishing an animal model of gestational diabetes mellitus as claimed in claim 3, characterized in that: The step 1) specifically includes: after 1 week of adaptive feeding, 11-week-old C57BL / 6J female mice are randomly divided into two groups, a CON group and a GDM group, with the same number of mice in each group, and the basal fasting blood glucose is measured.
5. Use of a hydrogel-loaded long-acting insulin receptor antagonist in establishing an animal model of gestational diabetes mellitus as claimed in claim 3, characterized in that: The step 2) specifically includes: after grouping, C57BL / 6J female mice are mated with male mice in a 2:1 ratio, and the vaginal plug is observed. The observation of the vaginal plug is recorded as pregnancy day 0.
6. Use of a hydrogel-loaded long-acting insulin receptor antagonist in establishing an animal model of gestational diabetes mellitus as claimed in claim 3, characterized in that: The step 3) is specifically as follows: preparing an injectable phenylboronic acid-hyaluronic acid hydrogel injection solution with phenylboronic acid-hyaluronic acid hydrogel and PBS buffer, wherein the ratio of the two is 1:4 (w:v, 1:4).
7. Use of a hydrogel-loaded long-acting insulin receptor antagonist in establishing an animal model of gestational diabetes mellitus as claimed in claim 3, characterized in that: The step 3) is specifically as follows: 1 mg of insulin receptor antagonist is added to 1 ml of PBS to dissolve to 206 nmol / mL, to prepare an insulin receptor antagonist stock solution, which is diluted 30 times with PBS buffer, and then mixed with phenylboronic acid-hyaluronic acid hydrogel in a ratio of 4:1, vortexed at room temperature, and sonicated for 10 minutes to allow the phenylboronic acid-hyaluronic acid hydrogel to fully load the insulin receptor antagonist.
8. Use of a hydrogel-loaded long-acting insulin receptor antagonist in establishing an animal model of gestational diabetes mellitus as claimed in claim 3, characterized in that: The step 4) is specifically as follows: when the C57BL / 6J female mice are 7 days pregnant, the fasting blood glucose is measured on the 7th day of pregnancy; starting from the 7th day of pregnancy of the C57BL / 6J female mice, according to the weight of the animals, the GDM group is subcutaneously injected with 60nmol / kg of insulin receptor antagonist polypeptide loaded with phenylboronic acid-hyaluronic acid hydrogel every 2 days to establish a gestational diabetes animal model, and the control group is injected with phenylboronic acid-hyaluronic acid hydrogel injection, and the injection volume is <150μl.
9. Use of a hydrogel-loaded long-acting insulin receptor antagonist in establishing an animal model of gestational diabetes mellitus as claimed in claim 3, characterized in that: In the step 4), the phenylboronic acid-hyaluronic acid hydrogel injection and the insulin receptor antagonist loaded by the phenylboronic acid-hyaluronic acid hydrogel are administered at 5 pm, once every 2 days until delivery.
10. Use of a hydrogel-loaded long-acting insulin receptor antagonist in establishing an animal model of gestational diabetes mellitus as claimed in claim 3, characterized in that: In step 5), after 7 days of administration, the pregnant mice with gestational diabetes and the control group were fasted for 10 hours, and blood was collected from the tail vein to determine the fasting blood sugar. Then, the pregnant mice were gavaged with glucose solution, and blood sugar was measured at 30 minutes, 60 minutes, 120 minutes, and 180 minutes after gavage, and an oral glucose tolerance test was performed.