A method for constructing a pituitary prolactinoma animal model
By subcutaneously injecting 17B-Estradiol into the back of animals to construct a pituitary prolactinoma model, the problem of the lack of stable models was solved, and a highly reproducible and low-cost experimental simulation was achieved, which is suitable for pituitary prolactinoma research and new drug screening.
Patent Information
- Application Number
- CN202510675132.2
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- Filing Date
- 2025-05-23
- Publication Date
- 2026-02-06
- Estimated Expiration
- 2045-05-23
AI Technical Summary
The lack of stable animal models of pituitary prolactinoma in current technology hinders research on pathogenesis and the development of new drugs.
Animal models of pituitary prolactinoma were established by subcutaneous injection of 17B-Estradiol into the back of both male and female animals. The solution used consisted of 20% DMSO, 30% anhydrous ethanol, and 50% glycerol, with a final concentration of 5.5 mg/ml. The drugs were administered every other day for 28 days.
The constructed model can simulate the pathophysiological process of human pituitary prolactinoma, with good reproducibility, low mortality, simple operation, and low cost, making it suitable for large-scale experiments and filling the gaps in existing models.
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Figure CN120459112B_ABST
Abstract
Description
TECHNICAL FIELD
[0001] The present application relates to the field of biotechnology, in particular to a method for constructing a pituitary prolactin adenoma animal model. BACKGROUND
[0002] Pituitary prolactin adenoma accounts for 30-40% of pituitary adenoma, is more common in 20-30 year-old women, and is often microadenoma. The typical clinical manifestations are amenorrhea-galactorrhea-infertility triad. Male prolactinoma is relatively rare, which is larger and often develops upward to the saddle. The main clinical manifestations are complete or partial sexual dysfunction and male infertility. If intratumoral hemorrhage occurs in the pituitary prolactin adenoma growing rapidly, acute pituitary apoplexy occurs, which is manifested as sudden severe headache, nausea, vomiting and visual impairment, even coma and exophthalmos, which requires emergency rescue. The patient may develop pituitary hypofunction after successful rescue.
[0003] There are various treatment options for pituitary prolactin adenoma in clinic, including surgery, radiotherapy and drug therapy. Endoscopic transsphenoidal pituitary tumor resection has become the main treatment for pituitary adenoma due to its definite effect, small trauma, clear vision, more extensive exposure, accurate operation, and rapid postoperative recovery. However, how to improve the prognosis of pituitary prolactin adenoma patients is still the most concerned topic in neurosurgery. However, adenoma is difficult to completely resect, and residual tumor tissue is prone to recurrence. In addition, some pituitary prolactin adenomas develop drug resistance, which has a poor prognosis and is a difficult problem to be solved in clinic.
[0004] At present, there is no stable pituitary prolactin adenoma experimental animal model, which seriously hinders the research on the pathogenesis of pituitary prolactin adenoma and the development of new drugs. Therefore, it is an urgent problem for those skilled in the art to provide a method for constructing a stable pituitary prolactin adenoma animal model. SUMMARY
[0005] In view of the deficiencies of the prior art, the present application provides a method for constructing a pituitary prolactin adenoma animal model, which constructs a pituitary prolactin adenoma animal model by subcutaneous injection of 17B-Estradiol preparation. The present application can better simulate the pathophysiological process of human pituitary prolactin adenoma, and has the advantages of good repeatability, significant effect, low mortality, simple operation method, safety, low experimental cost and suitability for large-scale animal experiments, which makes up for the defects of the existing animal model construction effect, and can be used for the pathogenesis research and new drug development of pituitary prolactin adenoma, and has good application prospect.
[0006] To achieve the above-mentioned purpose, the technical scheme adopted by the present application is as follows:
[0007] The first aspect of the present application provides a method for constructing a pituitary prolactinoma animal model, the steps of the method comprising: subcutaneously injecting 17B-Estradiol preparation into the back of the animals.
[0008] In the present application, 17B-Estradiol, also known as beta-estradiol, has a CAS number of 50-28-2, and is a female hormone secreted by the ovary. 17B-Estradiol is widely present in rivers, soil, water sources, atmosphere and agricultural products. It mainly comes from pesticides, exhaust gas, food additives, excrement of humans and animals, and domestic sewage, and can have a significant impact on organisms even at very low concentrations, and its content level is significantly correlated with prostate cancer, breast cancer, ovarian cancer, uterine cancer and male reproductive disorders.
[0009] Further, the steps of the method comprise: subcutaneously injecting 0.1 ml of 17B-Estradiol preparation into the back of the female animals every other day; and subcutaneously injecting 0.2 ml of 17B-Estradiol preparation into the back of the male animals every other day. The total duration of the method is 28 days.
[0010] Further, the configuration method of the 17B-Estradiol preparation is: adding 17B-Estradiol to a solvent; the composition of the solvent is 20% DMSO, 30% anhydrous ethanol and 50% glycerol.
[0011] Further, the final concentration of 17B-Estradiol in the 17B-Estradiol preparation is 5.5 mg / ml.
[0012] Further, the animals include one or more of mice, rabbits, dogs and monkeys.
[0013] In the present application, the animals are preferably mice, more preferably rats.
[0014] Further, the animals are 8-week-old rats.
[0015] Further, the animals are Wistar rats.
[0016] The second aspect of the present application provides the use of 17B-Estradiol and / or the 17B-Estradiol preparation described in the first aspect of the present application in constructing a pituitary prolactinoma animal model.
[0017] The third aspect of the present application provides the use of the animal model obtained based on the construction method described in the first aspect of the present application in screening drugs for treating pituitary prolactinoma.
[0018] The fourth aspect of the present application provides a method for screening a drug for treating pituitary prolactinoma, the method comprising the following steps:
[0019] administering the drug to be tested to the animal model obtained by the construction method of the first aspect of the present application;
[0020] analyzing the therapeutic effect of the drug to be tested and evaluating, and the drug to be tested which can obviously improve the pituitary prolactinoma of the animal model is a candidate drug for treating pituitary prolactinoma.
[0021] In some embodiments, the drug to be tested comprises natural drugs, traditional Chinese medicine monomers, small molecule compounds, and antibodies.
[0022] In some embodiments, the natural medicines, traditional Chinese medicine monomers, and small molecule compounds are sourced from: newly synthesized or existing databases; wherein existing databases include, but are not limited to, general natural product databases (COCONUT, SuperNatural II, NPASS), plant natural product databases (KNApSaCK, CMAUP, TriForC, Alkamid, NPACT DB, BioPhytMol), traditional Chinese medicine natural product databases (TCM@Taiwan, CEMTDD, CHDD, ETCM, TM-MC, TCMID, YaTCM), microbial natural product databases (StreptomeDB, NP Altas, ProCarDB, PAMDB, LichenDatabase), marine natural product databases (MNPD, SWMD), natural product databases from different countries and regions (IMPPAT, NeMedPlant, MedPServer, TlPdb, AfroDB, ANPDB, BIOFACQUIM, NUBBEDB), and food natural product databases (FooDB, BitterDB, Phenol-Explorer, PhytoHub, SuperSweet). Databases including: toxic natural product databases (Exposome-Explorer, T3DB, Snake Neurotoxin Database, TPPT), natural product industry catalogs (Greenpharma, AnalyticCon Discovery, InterBioScreen, Indofine Chemical Company, Pi Chemicals Systems\Specs, TargetMol), databases deduplicated using MS data (MoNA, MassBank, METLIN, HMDB, YMDB, ReSpect, GNPS), and databases deduplicated using NMR data (NMRShiftDB, NAPROC-13), etc.
[0023] In some embodiments, antibodies are used in the broadest sense in this invention and explicitly cover single-domain antibodies, monoclonal antibodies, polyclonal antibodies, human antibodies, chimeric antibodies, and multispecific antibodies (e.g., bivalent antibodies) formed from at least two intact antibodies, provided they exhibit the desired biological activity (e.g., therapeutic or alleviating effect on pituitary prolactinoma).
[0024] Advantages and beneficial effects of the present application: the animal model construction method provided by the present application can better simulate the pathophysiological process of human pituitary prolactin adenoma, has repeatability, remarkable effect, low mortality, simple and safe operation method, low experimental cost and the advantages of being suitable for large-scale animal experiments, makes up for the defects of the existing animal model construction effect, fills the gap in this research at home and abroad, can be used for the pathogenesis research and new drug development of pituitary prolactin adenoma, and has good application prospect. BRIEF DESCRIPTION OF DRAWINGS
[0025] Figure 1 Representative images of MRI sagittal and coronal of the constructed animal model.
[0026] Figure 2 Statistical graph of pituitary volume change of the constructed animal model rat.
[0027] Figure 3 Statistical graph of pituitary weight change of the constructed animal model rat.
[0028] Figure 4 Statistical graph of serum prolactin (PRL) level change of the constructed animal model rat.
[0029] Figure 5 Representative image of HE staining of the constructed animal model rat adenoma. DETAILED DESCRIPTION
[0030] The technical solutions in the embodiments of the present application will be described clearly and completely below in combination with the drawings in the embodiments of the present application. Obviously, the described embodiments are only part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor fall within the scope of protection of the present application.
[0031] In the technical solutions and experimental processes described in the present application, all kinds of reagents involved are common reagents or commercial reagents that can be clearly known and easily obtained by those skilled in the art based on their professional knowledge and conventional practice. The description of reagents in the present application is intended to clearly illustrate the material basis involved in the technical solutions, and those skilled in the art can successfully obtain and correctly use these reagents based on their professional accomplishment and industry common sense to achieve the technical purpose of the present application.
[0032] EMBODIMENT
[0033] 1. Experimental materials:
[0034] 8 weeks old Wistar female and male rats, female rats weighing 130-180±10 g, male rats weighing 180-230±10 g, were raised in a barrier environment, with a temperature of 22°C, humidity of 50%, 12-hour day / night rhythm, and fed with sterile feed and sterilized water.
[0035] Sterile feed formula: casein: 180-250 g / kg, soybean oil: 40-60 g / kg, corn starch: 300-400 g / kg, cellulose: 30-50 g / kg. Mineral premix: 30-40 g / kg, vitamin premix: 10-20 g / kg, containing vitamins A, D, E, K, and B vitamins, etc.
[0036] 2. Model construction method:
[0037] 1) Take 8-week-old female and male rats, 8 rats each, and randomly divide them into a control group and a model group, with 3 rats in the control group and 5 rats in the model group.
[0038] 2) The rats in the model group were given 17B-Estradiol preparation: 20% DMSO + 30% anhydrous ethanol + 50% glycerol + 5.5 mg / ml 17B-Estradiol (solid). The control group was given solvent: 20% DMSO + 30% anhydrous ethanol + 50% glycerol. 0.1 ml (female) / 0.2 ml (male) of 17B-Estradiol preparation / solvent was given every 48 hours, and the drug was administered subcutaneously on the back of the rats.
[0039] 3) The rats were given sterile feed and sterilized water every other day, and were raised for a total of 28 days.
[0040] 3. Detection of pituitary prolactinoma animal model construction:
[0041] 1) MRI sagittal and coronal scans were performed on the 1st day and the 28th day (results shown in Figure 1 ), and the rat pituitary volume was calculated using RadiAnt2025 version software (results shown in Figure 2 ). After 28 days, the rat pituitary was weighed and counted (results shown in Figure 3 ), and the detailed data are shown in Table 1.
[0042] 2) On the 1st day and the 28th day, the rats were anesthetized with isoflurane, and blood was taken from the inner canthus, and serum (100 μl) was taken after incubation with labeled antibodies at 4°C for 20 h, and separated with a separator for 15 min at 3800 rpm for 15 min. The serum was detected for serum prolactin (PRL) hormone level using an iodine [125I] prolactin radioimmunoassay kit (results shown in Figure 4 ), and the PRL level of the model group was significantly increased compared with the control group, indicating that the model was successful.
[0043] 3) Take the pituitary section of the rats on day 28 and perform HE staining, and the pathology shows pituitary prolactin adenoma, which is considered as successful modeling, and the results are shown in Table 2. Figure 5
[0044] Table 1. Detailed data of model group and control group rats
[0045]
[0046]
[0047] The above has been described in detail. For those skilled in the art, the present application can be implemented in a wider range under equivalent parameters, concentrations and conditions without departing from the purpose and scope of the present application and without unnecessary experiments. Although the present application gives examples, it should be understood that further improvements can be made to the present application. In summary, according to the principle of the present application, the present application is intended to include any change, use or improvement of the present application, including changes made by conventional techniques known in the art, which deviates from the range disclosed in the present application.
Claims
1. A method for constructing an animal model of pituitary prolactinoma, characterized in that, The construction method includes the following steps: subcutaneously injecting a 17B-Estradiol preparation into the back of male and female 8-week-old Wistar rats, wherein the final concentration of 17B-Estradiol in the 17B-Estradiol preparation is 5.5 mg / ml.
2. The construction method according to claim 1, characterized in that, The steps of the construction method include: 0.1 ml of 17B-Estradiol was subcutaneously injected into the back of the female animal, and administered every other day. 0.2 ml of 17B-Estradiol was subcutaneously injected into the back of male animals, with administration every other day; The total construction time of the method is 28 days.
3. The construction method according to claim 1, characterized in that, The preparation method of the 17B-Estradiol formulation is as follows: 17B-Estradiol is added to the solvent; The solvent consists of 20% DMSO, 30% anhydrous ethanol, and 50% glycerol.