Induction construction method of machin hypertension model

Abstract

The invention discloses an induced construction method of a machin hypertension model, which comprises the following steps: mixing a PDMS-PEG copolymer, DOCA-loaded PLGA microspheres, sodium alginate and triethyl citrate, adding a phosphate buffer solution, carrying out vortex oscillation to form homogeneous sol, and filtering and sterilizing to obtain injectable DOCA homogeneous sol; the prepared DOCA homogeneous sol is injected to cynomolgus monkeys according to the dosage of the injection volume of 0.5-1.0 mL / kg; after injection, the cynomolgus monkeys are fed with high-salt diet for 12-13 weeks, and molding is completed. Through the synergistic effect of the innovative DOCA homogeneous sol, the subcutaneous injection method and the high-salt diet, the problems of large animal trauma, high infection risk, long experimental period and the like caused by a traditional administration mode are successfully solved, and a more efficient, stable and accurate technical means is provided for construction of a hypertension model.

Description

Technical Field

[0001] The present invention relates to the technical field of disease animal model construction, and in particular to a method for inducing and constructing a cynomolgus monkey hypertension model. Background Art

[0002] The development of animal experimental models is crucial in the research of hypertension and related cardiovascular diseases. Animal experimental models bridge basic research with clinical application, providing indispensable tools for in-depth understanding of disease pathogenesis, identifying effective diagnostic methods, and developing new therapeutic strategies. By mimicking the onset and progression of human diseases, animal models allow researchers to observe the effects of disease on various body systems under relatively controlled conditions, investigate the role of various factors in disease progression, and conduct preliminary evaluations of potential treatments. For example, when studying the pathogenesis of hypertension, the development of appropriate animal models can reveal the specific roles of key components of elevated blood pressure, such as vasoconstriction, sodium and water retention, and renal dysfunction. In drug development, animal models can be used to screen and test the efficacy and safety of new antihypertensive drugs, providing important reference for clinical trials. Furthermore, animal models can facilitate the study of the pathophysiology of hypertension-induced cardiovascular complications, such as myocardial hypertrophy, heart failure, and atherosclerosis, thereby advancing the overall prevention and treatment of cardiovascular diseases. Therefore, the development of scientific, rational, and reliable animal experimental models is of irreplaceable importance for the in-depth development of research on hypertension and related cardiovascular diseases.

[0003] DOCA is a synthetic mineralocorticoid that can mimic the effects of aldosterone, promoting sodium reabsorption and potassium excretion, thereby increasing blood volume and blood pressure. A high-salt diet further aggravates sodium retention, synergizing with the effects of DOCA, leading to a significant increase in blood pressure. Currently, in terms of DOCA (deoxycorticosterone) administration, the traditional silicone rod implantation method is widely used, but it has significant problems. During the operation, the animal needs to undergo a large incision operation, which not only causes greater physical trauma to the animal and increases the risk of infection, but may also affect the accuracy of the experimental results due to surgical stress, prolong the animal's recovery time, and interfere with normal physiological metabolic processes. The traditional subcutaneous injection method requires repeated subcutaneous injections of DOCA oil or suspension, and the formation of hypertension takes a long time, thereby increasing the chance of infection in the animal and the labor intensity of the experimenter. Summary of the Invention

[0004] To address the aforementioned technical issues, the present invention aims to provide a method for inducing a hypertension model in cynomolgus monkeys. By combining an innovative DOCA homogenous sol with subcutaneous injection and a high-salt diet, this method successfully overcomes the significant animal trauma, high infection risk, and long experimental cycles associated with traditional drug administration methods, providing a more efficient, stable, and precise technical approach for constructing a hypertension model.

[0005] In order to achieve the above technical objectives and the above technical effects, the present invention is implemented through the following technical solutions:

[0006] A method for inducing and constructing a cynomolgus monkey hypertension model comprises the following steps:

[0007] (1) Preparation of DOCA homogeneous sol

[0008] PDMS-PEG copolymer, DOCA-loaded PLGA microspheres, sodium alginate, and triethyl citrate were mixed, phosphate buffer was added, and the solid content was adjusted to 20%-30%. The mixture was vortexed to form a homogeneous sol, and then filtered and sterilized to obtain an injectable DOCA homogeneous sol.

[0009] (2) Injection of DOCA homogeneous sol

[0010] The prepared DOCA homogenous sol was injected into cynomolgus monkeys at an injection volume of 0.5-1.0 mL / kg;

[0011] (3) Feeding a high-salt diet

[0012] After injection, cynomolgus monkeys were fed a high-salt diet for 12-13 weeks to complete the modeling. Furthermore, based on the total mass percentage of PDMS-PEG copolymer, DOCA-loaded PLGA microspheres, sodium alginate, and triethyl citrate as 100%, the mass percentage of PDMS-PEG copolymer was 70-80%, the mass percentage of DOCA-loaded PLGA microspheres was 10-25%, the mass percentage of sodium alginate was 1-3%, and the mass percentage of triethyl citrate was 0.5-2%.

[0013] Furthermore, in the PDMS-PEG copolymer, the molecular weight of PDMS is 900-2500 g / mol, the molecular weight of PEG is 400-1000 g / mol, and the molar ratio of PDMS to PEG is 3:4-4:5.

[0014] Furthermore, the preparation method of the DOCA-loaded PLGA microspheres is:

[0015] DOCA and PLGA were dissolved in a solvent at a certain mass ratio and ultrasonically emulsified to form an emulsion. The emulsion was added dropwise to the PVA solution, and the solvent was evaporated under reduced pressure under magnetic stirring. The microspheres were collected by centrifugation and freeze-dried for storage.

[0016] Furthermore, the drug loading amount in the DOCA-loaded PLGA microspheres is 10-20%.

[0017] Furthermore, the cynomolgus monkeys are healthy cynomolgus monkeys aged 3-5 years and weighing 5-10 kg, and the basic blood pressure screening is: systolic pressure <140 mmHg, diastolic pressure <90 mmHg.

[0018] Furthermore, the DOCA homogeneous sol is injected subcutaneously into the back or abdomen.

[0019] Furthermore, in step (3), the feeding cycle of the high-salt diet includes an induction period and a maintenance period. The induction period adopts a high-sodium, low-potassium diet, and the maintenance period adopts a step-by-step reduction in sodium content.

[0020] Furthermore, the high-salt diet is a basic high-salt feed or a high-salt and high-fat feed.

[0021] Furthermore, the basic high-salt feed includes 3-8% sodium chloride, 1.0-1.2% potassium chloride, and the remaining ingredients are standard primate maintenance feed;

[0022] The high-salt and high-fat feed comprises the following components in percentage by mass: 3%-8% sodium chloride, 0.2%-1% KCl, 15%-25% lard or tallow, 0.2%-2% cholesterol, 0.02%-0.2% bile salt, 20%-30% corn starch, 20%-30% casein, 10%-15% sucrose, 2%-5% microcrystalline cellulose, 1%-1.2% calcium dihydrogen phosphate, and 3%-5% vitamins.

[0023] Furthermore, an intelligent feeding system was used to feed a high-salt diet, which recorded daily food intake to control sodium intake error to ≤2%, and a blood pressure monitoring device was used to record the systolic and diastolic blood pressure of the crab-eating macaques.

[0024] The beneficial effects of the present invention are:

[0025] The present invention can effectively induce a state of hypertension in crab-eating macaques through a comprehensive approach of injecting a DOCA homogenous sol combined with feeding a high-salt diet. Specifically, the present invention prepares a homogenous sol by loading DOCA into PLGA microspheres and mixing it with a PDMS-PEG copolymer, sodium alginate, and triethyl citrate. The homogenous sol is liquid before injection, and after being injected into the crab-eating macaque, cross-linking is triggered by body temperature, rapidly forming a gel-like structure. This allows the release rate of DOCA to be controlled, achieving long-term and stable drug release, allowing DOCA to continue to exert its effect in the crab-eating macaque, better simulating the occurrence and development process of hypertension, and improving the stability and success rate of model construction. DOCA and a high-salt diet work together to simulate the occurrence process of human hypertension, forming a relatively stable hypertension model that conforms to physiological and pathological characteristics.

[0026] The present invention prepares an injectable DOCA homogeneous sol and adopts a subcutaneous injection method of the back or abdomen, thereby avoiding the large-incision surgery of the traditional silicone rod implantation method and reducing the physical trauma and infection risk of the animal. At the same time, a single injection of the DOCA homogeneous sol can continuously release DOCA without the need for repeated injections, which not only reduces the probability of animal infection but also enables the construction of a hypertension model in a shorter time, shortens the experimental cycle, and improves the drug administration efficiency. At the same time, the frequency of repeated injections by experimenters is reduced, the labor intensity is reduced, and the convenience of the experiment is improved.

[0027] DOCA in this invention activates the mineralocorticoid receptor in the renal tubules, promoting sodium reabsorption and potassium excretion, leading to sodium retention, increased blood volume, and, in turn, elevated blood pressure. A high-salt diet increases the concentration of sodium ions in the body, activating the sodium-potassium-ATPase enzyme in the kidneys, further promoting sodium reabsorption and potassium excretion. The synergistic effect of DOCA and a high-salt diet can rapidly induce a significant increase in blood pressure in cynomolgus monkeys, enabling the efficient construction of a hypertension model.

[0028] The advantages of using cynomolgus macaques to construct a hypertension model in this invention are: their genetic and physiological characteristics are highly similar to those of humans, accurately reflecting the pathogenesis and physiological and pathological characteristics of human hypertension. Their blood pressure regulation mechanisms are similar to those of humans, and the model is stable and reproducible, making it suitable for long-term experimental observations and drug efficacy evaluation. Furthermore, cynomolgus macaques are amenable to a variety of experimental procedures, meeting diverse research needs. They are particularly valuable in preclinical drug and vaccine research, making research results more easily translated into clinical applications and potentially aiding the prevention, diagnosis, and treatment of hypertension. DETAILED DESCRIPTION

[0029] The technical solutions of the present invention are described clearly and completely below with reference to specific embodiments. Obviously, the embodiments described are only some embodiments of the present invention, not all embodiments. All other embodiments derived by persons of ordinary skill in the art based on the embodiments of the present invention without creative effort are also within the scope of protection of the present invention.

[0030] The present invention provides a method for inducing and constructing a cynomolgus monkey hypertension model, comprising the following steps:

[0031] (1) Preparation of DOCA homogeneous sol

[0032] A PDMS-PEG copolymer, DOCA-loaded PLGA microspheres, sodium alginate, and triethyl citrate are mixed, a phosphate buffer is added, the solid content is adjusted to 20%-30%, the mixture is vortexed to form a homogeneous sol, and the mixture is filtered and sterilized to prepare an injectable DOCA homogeneous sol; wherein, based on the total mass percentage of the PDMS-PEG copolymer, DOCA-loaded PLGA microspheres, sodium alginate, and triethyl citrate being 100%, the mass percentage of the PDMS-PEG copolymer is 70-80%, the mass percentage of the DOCA-loaded PLGA microspheres is 10-25%, the mass percentage of the sodium alginate is 1-3%, and the mass percentage of the triethyl citrate is 0.5-2%.

[0033] The preparation method of PLGA microspheres loaded with DOCA is as follows:

[0034] DOCA and PLGA were dissolved in a solvent (dichloromethane) at a mass ratio of 1:3-1:5, and ultrasonic emulsification (power 200W, time 5 minutes) was used to form an emulsion; the emulsion was added dropwise to a 1% PVA solution, and the solvent was evaporated under reduced pressure under magnetic stirring for 4-6 hours. The microspheres were collected by centrifugation, washed three times with deionized water, and freeze-dried to obtain PLGA microspheres with a drug loading of 10-20% and a particle size of 80-120 nm.

[0035] In the PDMS-PEG copolymer, the PDMS molecular weight is 900-2500g / mol, the PEG molecular weight is 400-1000g / mol, and the PDMS to PEG molar ratio is 3:4-4:5. The PDMS-PEG copolymer is thermosensitive. Under body temperature, the mobility of the PDMS segments is enhanced, while the PEG segments interact with other molecules through intermolecular forces (such as hydrogen bonds and van der Waals forces). This heat-induced segment motion and interaction causes the system to transform from a flowing homogeneous sol to an elastic gel network. Sodium alginate can produce certain crosslinks with the PDMS-PEG copolymer segments, further enhancing the overall network structure and making the gel more stable. The presence of triethyl citrate helps to regulate the compatibility and flexibility of the system, allowing the various components to better fuse with each other, reducing the possibility of phase separation, and thus facilitating the formation of a uniform and stable gel system. DOCA-loaded PLGA microspheres are encapsulated within the resulting gel network. The sustained-release properties of the microspheres allow for a gradual release of DOCA; the gel network further slows the drug's diffusion rate, achieving a more sustained release effect. Under the combined influence of the in vivo environment, the entire system solidifies into a stable gel through physical cross-linking and intermolecular interactions, achieving long-term sustained release of the drug.

[0036] (2) Injection of DOCA homogeneous sol

[0037] The prepared DOCA homogenous sol was injected into cynomolgus monkeys at an injection volume of 0.5-1.0 mL / kg;

[0038] Among them, the screening criteria for crab-eating macaques are: age 3-5 years old, weight 5-10kg, basal systolic blood pressure <140mmHg, diastolic blood pressure <90mmHg; CRISPR-Cas9 gene editing can also be used to screen out individuals with congenital aldosterone synthase (CYP11B2) deficiency.

[0039] (3) Feeding a high-salt diet

[0040] After injection, the cynomolgus monkeys were fed a high-salt diet for 12-13 weeks to complete the modeling.

[0041] In step (3), the feeding cycle of the high-salt diet includes an induction period (weeks 1-4) and a maintenance period (weeks 5-13). During the induction period, a high-sodium, low-potassium diet is adopted. During the maintenance period, the sodium content is stepwise reduced to 1.8-2.5%, and potassium is supplemented to 1.5-2.0%.

[0042] The high-salt diet can be selected from a basic high-salt feed or a high-salt, high-fat feed. Preferably, the basic high-salt feed comprises 3-8% sodium chloride, 1.0-1.2% potassium chloride, and the remaining ingredients are standard primate maintenance feed; the high-salt, high-fat feed comprises the following components by mass percentage: 3%-8% sodium chloride, 0.2-1% KCl, 15%-25% lard or tallow, 0.2%-2% cholesterol, 0.02%-0.2% bile salt, 20%-30% corn starch, 20%-30% casein, 10%-15% sucrose, 2-5% microcrystalline cellulose, 1%-1.2% calcium dihydrogen phosphate, and 3%-5% vitamins.

[0043] An intelligent feeding system is used to feed a high-salt diet, record daily food intake to control sodium intake error ≤ 2%, and record the systolic and diastolic blood pressure of the crab-eating macaques using a blood pressure monitoring device. Specifically, the blood pressure monitoring method is: a wireless PPG sensor is fixed to the tail or ear of the crab-eating macaque to continuously monitor the blood pressure waveform (sampling frequency 100Hz). The tail cuff method (a special tail cuff is placed at the base of the animal's tail, and the tail artery is pressurized and released by inflation and deflation, while the blood flow signal of the tail artery is monitored) is used once a week to measure and calibrate the PPG data to ensure an error of <±5mmHg.

[0044] The high-salt diet in this invention is designed to be fed in an induction phase and a maintenance phase. The induction phase uses a high-sodium, low-potassium diet, while the maintenance phase involves a stepwise reduction in sodium content. This allows for more precise control of sodium intake, synergistically with the sustained release of DOCA to further aggravate sodium retention, more effectively inducing a significant increase in blood pressure and optimizing the development of a hypertension model. Furthermore, the combination of an intelligent feeding system and a blood pressure monitoring device allows for precise control of sodium intake and real-time monitoring of blood pressure changes.

[0045] Preparation Example

[0046] Preparation of DOCA homogeneous sol

[0047] 75% PDMS-PEG copolymer, 22% DOCA-loaded PLGA microspheres, 2% sodium alginate, and 1% triethyl citrate were mixed, phosphate buffer was added, and the solid content was adjusted to 25%. The mixture was vortexed to form a homogeneous sol, and then filtered and sterilized to obtain an injectable DOCA homogeneous sol.

[0048] The preparation method of PLGA microspheres loaded with DOCA is as follows:

[0049] DOCA and PLGA were dissolved in dichloromethane at a mass ratio of 1:3, and ultrasonic emulsification (power 200 W, time 5 min) was used to form an emulsion. The emulsion was added dropwise to a 1% PVA solution, and the solvent was evaporated under reduced pressure under magnetic stirring for 5 hours. The microspheres were collected by centrifugation, washed three times with deionized water, and freeze-dried to obtain PLGA microspheres with a drug loading of 22% and a particle size of 100 nm.

[0050] Experimental group

[0051] Selection of cynomolgus monkeys: Healthy cynomolgus monkeys aged 3-5 years and weighing 5-10 kg, with baseline systolic blood pressure <140 mmHg and diastolic blood pressure <90 mmHg, and individuals with congenital aldosterone synthase (CYP11B2) deficiency were excluded by CRISPR-Cas9 gene editing screening.

[0052] Experimental steps:

[0053] Preparation and injection of DOCA homogenous sol

[0054] Preparation of DOCA homogeneous sol: 75% PDMS-PEG copolymer (PDMS molecular weight 1500 g / mol, PEG molecular weight 700 g / mol, PDMS to PEG molar ratio 4:5), 22% DOCA-loaded PLGA microspheres (drug loading 22%), 2% sodium alginate, and 1% triethyl citrate were mixed, phosphate buffer was added, and the solid content was adjusted to 25%. The mixture was vortexed to form a homogeneous sol, and then filtered and sterilized to obtain an injectable DOCA homogeneous sol.

[0055] Injection of DOCA homogenous sol: Administer 0.8 mL / kg of DOCA homogenous sol subcutaneously to the back of cynomolgus monkeys.

[0056] Feeding a high-salt diet

[0057] High-salt diet: During the induction period (weeks 1-4), a high-sodium, low-potassium diet was adopted (a high-salt, high-fat feed was used, and the component ratio was: the high-salt, high-fat feed included the following components in mass percentage: sodium chloride 5%, KCl 1%, lard 25%, cholesterol 1%, bile salt 1%, corn starch 25%, casein 25%, sucrose 10%, microcrystalline cellulose 3%, calcium dihydrogen phosphate 1%, vitamin 3%). During the maintenance period (weeks 5-13), the sodium content was reduced in a step-by-step manner (the sodium chloride content in the above feed was reduced from 5% → 3.5% → 2.5%, and the KCl content was increased simultaneously).

[0058] Feeding system: Use an intelligent feeding system to record daily food intake and control the sodium intake error to ≤2%.

[0059] Blood pressure monitoring

[0060] Monitoring Method: A wireless PPG sensor was attached to the tail of cynomolgus monkeys to continuously monitor blood pressure waveforms (sampling frequency 100 Hz). PPG data were calibrated using a tail cuff once a week to ensure an error of <±5 mmHg.

[0061] Monitoring frequency: Monitor blood pressure three times a week and record systolic blood pressure, diastolic blood pressure and heart rate data.

[0062] control group

[0063] Selection of cynomolgus monkeys: The same as the experimental group, healthy cynomolgus monkeys aged 3-5 years and weighing 5-10 kg, with baseline blood pressure systolic pressure <140 mmHg and diastolic pressure <90 mmHg, were screened by CRISPR-Cas9 gene editing to exclude individuals with congenital aldosterone synthase (CYP11B2) deficiency.

[0064] Experimental steps:

[0065] Injection of blank sol

[0066] Preparation of blank sol: 75% PDMS-PEG copolymer (PDMS molecular weight 1500 g / mol, PEG molecular weight 700 g / mol, PDMS to PEG molar ratio 4:5), 22% blank PLGA microspheres (without DOCA), 2% sodium alginate, and 1% triethyl citrate were mixed, phosphate buffer was added, and the solid content was adjusted to 25%. The mixture was vortexed to form a homogeneous blank sol, and then filtered and sterilized to obtain an injectable blank sol.

[0067] Injection of blank sol: Administer a subcutaneous injection into the back of cynomolgus monkeys at a dose of 0.8 mL / kg.

[0068] Feeding a normal diet

[0069] Normal diet: Standard primate maintenance chow without additional high-salt ingredients.

[0070] Feeding system: Use intelligent feeding system to record daily food intake.

[0071] Blood pressure monitoring

[0072] Monitoring Method: A wireless PPG sensor was attached to the tail of cynomolgus monkeys to continuously monitor blood pressure waveforms (sampling frequency 100 Hz). PPG data were calibrated using a tail cuff once a week to ensure an error of <±5 mmHg.

[0073] Monitoring frequency: Monitor blood pressure three times a week and record systolic blood pressure, diastolic blood pressure and heart rate data.

[0074] The experimental and control groups were observed for 13 weeks, with weekly systolic and diastolic blood pressure and heart rate data recorded. The blood pressure trends of the experimental and control groups were compared. The blood pressure monitoring results for the experimental and control groups are shown in Table 1.

[0075] Table 1

[0076]

[0077] The data in Table 1 are expressed as mean ± standard deviation; *P < 0.05, **P < 0.01, ***P < 0.001, indicating that there are significant differences between the experimental group and the control group.

[0078] As shown in Table 1, the systolic and diastolic blood pressures of the experimental group gradually increased during the experiment, indicating that DOCA homogeneous sol combined with a high-salt diet can effectively induce hypertension; the heart rate of the experimental group gradually increased, which is related to the increased cardiac load caused by increased blood pressure.

[0079] Cardiac ultrasound and vascular function assessment

[0080] Echocardiography is used to assess the effects of hypertension on cardiac structure and function; the methods are:

[0081] Sample collection: At the end of the experiment (week 13), all cynomolgus monkeys underwent cardiac ultrasound examination.

[0082] Check indicators:

[0083] Left ventricular mass index (LVMI): assesses the degree of left ventricular hypertrophy;

[0084] Left ventricular ejection fraction (LVEF): assesses left ventricular systolic function;

[0085] Left ventricular end-diastolic diameter (LVEDD): assesses left ventricular diastolic function;

[0086] Left atrial diameter (LAD): Assess the degree of left atrial dilation.

[0087] The successful construction of the hypertension model was further verified by evaluating vascular function, especially endothelial function; the method was as follows:

[0088] Sample collection: At the end of the experiment (week 13), vascular function assessment was performed on all cynomolgus monkeys.

[0089] Check indicators:

[0090] Endothelium-dependent dilation (EDDF): assesses endothelial function by measuring acetylcholine-induced vasodilation;

[0091] Non-endothelium-dependent dilation (NEDDF): assesses vascular smooth muscle function by measuring nitroglycerin-induced vasodilation.

[0092] The evaluation results are shown in Table 2.

[0093]

[0094]

[0095] The data in Table 2 are expressed as mean ± standard deviation.

[0096] As shown in Table 2, the left ventricular mass index (LVMI) in the experimental group was significantly higher than that in the control group, indicating left ventricular hypertrophy, a typical manifestation of cardiac structural changes caused by hypertension. The left ventricular ejection fraction (LVEF) decreased slightly, while the left ventricular end-diastolic diameter (LVEDD) and left atrial diameter (LAD) increased significantly, indicating that cardiac function was affected to a certain extent. Both the endothelium-dependent dilation function (EDDF) and the non-endothelium-dependent dilation function (NEDDF) in the experimental group were significantly lower than those in the control group, indicating impaired endothelial function and reduced vascular smooth muscle reactivity, indicating that hypertension causes vascular dysfunction. These results indicate that the experimental group successfully established a hypertension model.

[0097] It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above and that the invention can be embodied in other specific forms without departing from the spirit or essential characteristics of the invention. Therefore, the embodiments should be considered in all respects as illustrative and non-restrictive, and the scope of the invention is defined by the appended claims rather than the foregoing description, and it is intended that all variations that come within the meaning and range of equivalents of the claims be embraced therein.

[0098] In addition, it should be understood that although this specification is described in terms of implementation methods, not every implementation method contains only one independent technical solution. This narrative method of the specification is only for the sake of clarity. Those skilled in the art should regard the specification as a whole. The technical solutions in each embodiment can also be appropriately combined to form other implementation methods that can be understood by those skilled in the art.

Claims

1. A method for inducing and constructing a cynomolgus monkey hypertension model, characterized in that: The following steps are involved: (1) Preparation of DOCA homogeneous sol PDMS-PEG copolymer, DOCA-loaded PLGA microspheres, sodium alginate, and triethyl citrate were mixed, phosphate buffer was added, and the solid content was adjusted to 20%-30%. The mixture was vortexed to form a homogeneous sol, and then filtered and sterilized to obtain an injectable DOCA homogeneous sol. (2) Injection of DOCA homogeneous sol The prepared DOCA homogenous sol was injected into cynomolgus monkeys at an injection volume of 0.5-1.0 mL / kg; (3) Feeding a high-salt diet After injection, the cynomolgus monkeys were fed a high-salt diet for 12-13 weeks to complete the modeling.

2. The method for inducing and constructing a cynomolgus monkey hypertension model according to claim 1, characterized in that: Based on the total mass percentage of PDMS-PEG copolymer, DOCA-loaded PLGA microspheres, sodium alginate, and triethyl citrate being 100%, the mass percentage of PDMS-PEG copolymer is 70-80%, the mass percentage of DOCA-loaded PLGA microspheres is 10-25%, the mass percentage of sodium alginate is 1-3%, and the mass percentage of triethyl citrate is 0.5-2%.

3. The method for inducing and constructing a cynomolgus monkey hypertension model according to claim 1, characterized in that: The preparation method of the DOCA-loaded PLGA microspheres is as follows: DOCA and PLGA were dissolved in a solvent at a certain mass ratio and ultrasonically emulsified to form an emulsion. The emulsion was added dropwise to the PVA solution, and the solvent was evaporated under reduced pressure under magnetic stirring. The microspheres were collected by centrifugation and freeze-dried for storage.

4. The method for inducing and constructing a cynomolgus monkey hypertension model according to claim 1, characterized in that: The drug loading amount in the DOCA-loaded PLGA microspheres is 10-20%.

5. The method for inducing and constructing a cynomolgus monkey hypertension model according to claim 1, characterized in that: The cynomolgus monkeys are healthy cynomolgus monkeys aged 3-5 years and weighing 5-10 kg, and the basic blood pressure screening is: systolic pressure <140 mmHg, diastolic pressure <90 mmHg.

6. The method for inducing and constructing a cynomolgus monkey hypertension model according to claim 1, characterized in that: DOCA homogeneous sol is injected subcutaneously in the back or abdomen.

7. The method for inducing and constructing a cynomolgus monkey hypertension model according to claim 1, characterized in that: In step (3), the feeding cycle of the high-salt diet includes an induction period and a maintenance period. During the induction period, a high-sodium, low-potassium diet is adopted, and during the maintenance period, the sodium content is stepwise reduced.

8. The method for inducing and constructing a cynomolgus monkey hypertension model according to claim 1, characterized in that: The high-salt diet is a basic high-salt feed or a high-salt and high-fat feed.

9. The method for inducing and constructing a cynomolgus monkey hypertension model according to claim 8, characterized in that: The basic high-salt feed includes 3-8% sodium chloride, 1.0-1.2% potassium chloride, and the remaining ingredients are standard primate maintenance feed; The high-salt and high-fat feed comprises the following components in percentage by mass: 3%-8% sodium chloride, 0.2%-1% KCl, 15%-25% lard or tallow, 0.2%-2% cholesterol, 0.02%-0.2% bile salt, 20%-30% corn starch, 20%-30% casein, 10%-15% sucrose, 2%-5% microcrystalline cellulose, 1.0%-1.2% calcium dihydrogen phosphate, and 3%-5% vitamin premix.

10. The method for inducing and constructing a cynomolgus monkey hypertension model according to claim 1, characterized in that: An intelligent feeding system was used to feed the macaques a high-salt diet, record daily food intake, control sodium intake error to ≤2%, and record the systolic and diastolic blood pressure of the macaques using a blood pressure monitoring device.

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