A rat model of benign prostatic hyperplasia, a method for establishing the same and use thereof

Abstract

The application discloses a rat benign prostatic hyperplasia model, a building method and application thereof, and belongs to the technical field of animal models. The building method comprises the following steps: D-galactose and testosterone undecanoate are jointly administered to rats in adaptive feeding to induce benign prostatic hyperplasia; the induction time is 60 days, D-galactose with a dosage of 125mg / kg / d-500mg / kg / d is continuously given to the rats by subcutaneous injection within 60 days, and testosterone undecanoate olive oil solution with a dosage of 15mg / kg / 10d is given to the rats by intraperitoneal injection starting from the 30th day. The prostate index of the rat benign prostatic hyperplasia model induced by D-galactose and testosterone undecanoate, serum hormone levels, histopathology, androgen receptor and proliferation-related molecules and testicular function are detected to comprehensively evaluate the effectiveness and practicability of the model, and the results show that the rat prostatic hyperplasia model induced by aging and androgen in the application is more close to the pathogenic characteristics of clinical benign prostatic hyperplasia.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of animal models, in particular to a benign prostatic hyperplasia model of rats, a method for establishing the same and application thereof. BACKGROUND

[0002] Benign prostatic hyperplasia (BPH) is a common disease in middle-aged and elderly men, mainly manifested as histological prostatic interstitial and glandular component hyperplasia, anatomical prostatic enlargement and bladder outlet obstruction (BOO) in urodynamics, and clinical symptoms such as lower urinary tract symptoms (LUTS) and related complications such as urinary frequency, urinary urgency, urinary incontinence, hesitancy, difficulty or interruption during urination, and dripping after urination. Most men over 65 years old have histological prostatic hyperplasia, of which about 2 / 3 have clinical symptoms.

[0003] With the deepening of the global population aging, the prevalence of benign prostatic hyperplasia is showing a sustained upward trend. As of now, the treatment of the disease is mainly drug maintenance and transurethral resection of the prostate (TURP), but due to long-term unstable drug efficacy, high risk of surgery for elderly patients and other reasons, there is still no exact and effective treatment. Therefore, an accurate preclinical animal model that can accurately reveal the molecular mechanism of benign prostatic hyperplasia and explore the treatment plan is indispensable.

[0004] Rat animal models, with their advantages of pure species, large quantity and low price, have become an important carrier for studying the pathogenesis of benign prostatic hyperplasia and evaluating the efficacy of drugs. In recent years, the establishment of benign prostatic hyperplasia rat models mainly includes the following:

[0005] (1) Testosterone induction: Since prostate growth is mainly regulated by androgens, the most common modeling method is the high-dose exogenous androgen induction method, which intervenes in normal or castrated (orchiectomy) male rats. This model can cause hyperplasia of the prostate epithelial layer and stromal interstitial space by disrupting the balance of endogenous androgens. Among them, the non-castration method has the advantages of simple operation and low infection rate, but it is easily affected by the regulation of its own hormones, and the degree of prostate hyperplasia and the maintenance time of the model are not ideal. The castration method has a higher modeling rate, and can cause a large degree of hyperplasia of the gland and interstitium in histopathology. However, testosterone is the most abundant and important androgen in human males, and is produced entirely by Leydig cells in the testes. With age, although the testicular function is weakened, it can cause a decrease in endogenous testosterone levels in the body, but the prostate and other androgen-dependent organs are different in that their receptors have the ability to respond to androgens throughout life.

[0006] Clinical studies have found that the level of dihydrotestosterone (DHT), a derivative of testosterone, and its receptor in the prostate of elderly men remains high. The testes maintain the production of DHT in the prostate by continuously providing testosterone, which is a key driving factor for the occurrence of benign prostatic hyperplasia. Therefore, androgen deprivation therapy has certain limitations: on the one hand, the surgical operation is difficult, and it is difficult to restore the prostate to the level before castration, and the efficacy evaluation is limited; on the other hand, it fails to reflect the characteristics of aging and the existence of testicular function in the pathogenesis of benign prostatic hyperplasia, and it is difficult to objectively reflect the core pathological changes of benign prostatic hyperplasia.

[0007] (2) Synergistic induction of estrogen and androgen: Studies have shown that with age, the prostate volume gradually increases, and the ratio of serum estrogen and androgen shows an upward trend. Therefore, estrogen and androgen synergistic intervention in castrated or non-castrated rats can exacerbate the imbalance of endogenous estrogen and androgen ratio leading to benign prostatic hyperplasia. This model has a high degree of agreement with the clinical diagnostic criteria, but the ratio of estrogen and androgen is often difficult to accurately evaluate, which affects the modeling rate to some extent.

[0008] (3) Urogenital sinus implantation method: Some scholars implant the urogenital sinus tissue of fetal rats of the same strain into the ventral lobe of the prostate to stimulate the proliferation ability of the embryonic tissue around the prostate urethra of the model animal, and induce benign prostatic hyperplasia. However, this method has the problems of high difficulty in operation, high risk of infection, and high animal mortality.

[0009] (4) Metabolic syndrome model: A number of preclinical and clinical research results have shown that metabolic abnormalities are an important determinant of the occurrence and progression of benign prostatic hyperplasia / LUTS. Therefore, abnormal morphology and function of the prostate are often observed in metabolic syndrome animal models such as high-fat diet and spontaneous hypertensive rats. However, this method is only suitable for exploring the relationship between benign prostatic hyperplasia and metabolic diseases, and has a narrow application scenario and a low modeling rate.

[0010] Aging and functional testes are two important factors recognized in the pathogenesis of clinical benign prostatic hyperplasia, but there is no benign prostatic hyperplasia model that takes both factors into account. The present application constructs a benign prostatic hyperplasia model induced by aging inducer D-galactose and testosterone undecanoate. The model introduces the aging factor to weaken the endocrine regulation of the testes and enhance the sensitivity of the prostate to exogenous androgens. The effectiveness and practicability of the model are comprehensively evaluated by changes in prostate index, serum hormone level, histopathology, androgen receptor, proliferation-related molecules, and testicular function. SUMMARY

[0011] The application aims to provide a rat benign prostatic hyperplasia model, a construction method and application thereof, so as to solve the problem that the existing model cannot accurately simulate the clinical pathogenic characteristics.

[0012] To achieve the above-mentioned object, the application provides a construction method of a rat benign prostatic hyperplasia model, wherein a D-galactose and an androgen testosterone undecanoate are jointly administered to an adaptive feeding rat to induce benign prostatic hyperplasia.

[0013] The induction time is 60 days, and the D-galactose is continuously administered at 125 mg / kg / d to 500 mg / kg / d by subcutaneous injection within 60 days, and the testosterone undecanoate is administered at 15 mg / kg / 10d by intraperitoneal injection starting from the 30th day.

[0014] Preferably, the rat is a Wistar rat.

[0015] Preferably, the adaptive feeding condition is that the rat is fed in a barrier environment, the light / dark cycle is 12h, the temperature is 25℃, the humidity is 75%, and the adaptive feeding lasts for 7 days.

[0016] Preferably, the subcutaneous injection dose of the D-galactose is 125 mg / kg / d.

[0017] Preferably, the subcutaneous injection dose of the D-galactose is 250 mg / kg / d.

[0018] Preferably, the subcutaneous injection dose of the D-galactose is 500 mg / kg / d.

[0019] In another aspect, the application provides an evaluation method of the above-mentioned rat benign prostatic hyperplasia model, which comprises at least one of the following evaluation contents:

[0020] (1) physiological indexes: prostate index, testis index and epididymal tail sperm count;

[0021] (2) serum hormone level: serum testosterone, dihydrotestosterone, free testosterone and prostate specific antigen concentration;

[0022] (3) histopathology: HE staining is used to observe the prostate gland structure and the testis spermatogenic tubule state, and SA-beta-gal staining is used to detect the prostate tissue senescent cells;

[0023] (4) molecular marker: the immunohistochemical method is used to detect the positive cell area percentage of the prostate tissue androgen receptor, proliferation marker Ki67 and PCNA.

[0024] In another aspect, the application provides an application of the above-mentioned rat benign prostatic hyperplasia model in the pathogenesis research of benign prostatic hyperplasia.

[0025] In another aspect, the present application provides a use of the above-mentioned rat benign prostatic hyperplasia model in the evaluation of drug screening for benign prostatic hyperplasia.

[0026] Therefore, the rat benign prostatic hyperplasia model, the establishing method and the application thereof have the following beneficial effects:

[0027] (1) The rat benign prostatic hyperplasia model is induced by the aging inducer D-galactose and testosterone undecanoate, and the effectiveness and practicability of the model are comprehensively evaluated through the changes of the prostate index, serum hormone level, histopathology, androgen receptor, proliferation-related molecules and testicular function, etc. The results show that the rat prostatic hyperplasia model induced by the combination of aging and androgen is more close to the pathogenic characteristics of clinical benign prostatic hyperplasia;

[0028] (2) In the process of evaluating the rat model, it is found that the single aging inducer D-galactose cannot cause rat prostatic hyperplasia, and the stability of the rat prostatic hyperplasia model induced by the single androgen testosterone undecanoate is poor;

[0029] (3) The prostate lateral lobe or posterior lobe index of the rats is analyzed, and it is found that the prostate lateral lobe of the rats in different administration groups or different dose administration groups is not different, and the prostate posterior lobe index of the rats in the D-galactose and testosterone undecanoate combined administration group is significantly increased compared with the blank treatment group, which is more similar to the peripheral structure prone to disease in humans;

[0030] (4) The different forms of testosterone in the serum of the rats in each group are detected, and the results show that the abundance of T, F-TESTO and DHT in each group of rats treated with testosterone undecanoate is increased to different degrees, and the prostate specific antigen (PSA) level is significantly increased in the rats in the testosterone undecanoate group and the high-dose group of the D-galactose and testosterone undecanoate combined model. At the same time, histopathological staining shows that after modeling, the mucosal folds of the prostate gland of the rats in the medium-dose group of the D-galactose and testosterone undecanoate combined model and the high-dose group of the D-galactose and testosterone undecanoate combined model are prominent, the glandular epithelial cells are hyperplastic, and the inflammatory infiltration is significant, and a large number of SA-β-Gal positive staining can be observed. Therefore, the rat benign prostatic hyperplasia model induced by the combination of D-galactose and testosterone undecanoate is more consistent with the pathogenic characteristics of clinical benign prostatic hyperplasia, and the modeling effect is better;

[0031] (5) The AR, Ki67 and PCNA levels in the prostate tissue of the rats were detected by immunohistochemical method. The results showed that the AR in the D-galactose and testosterone undecanoate combined model middle dose group and the D-galactose and testosterone undecanoate combined model high dose group was significantly increased after modeling. Meanwhile, compared with the normal group, the Ki67 and PCNA positive cell staining percentages in the D-galactose and testosterone undecanoate combined model middle dose group, the D-galactose and testosterone undecanoate combined model high dose group, the testosterone undecanoate group, the D-galactose and testosterone undecanoate combined model middle dose group and the D-galactose and testosterone undecanoate combined model high dose group were significantly increased, which indicated that the D-galactose and testosterone undecanoate combined model could cause the excessive proliferation of prostate cells through Ki67 and PCNA proteins;

[0032] (6) The testis index and sperm count were calculated, and it was found that the testis index and sperm count of the rats treated with the aging inducer D-galactose were significantly reduced. Meanwhile, the histopathological detection could directly show that aging could cause the damage of testis tissue cells. It was indicated that aging and androgen could cause the damage of the testis structure of the rats.

[0033] The technical solutions of the present application will be further described below by means of the accompanying drawings and examples. BRIEF DESCRIPTION OF DRAWINGS

[0034] In order to more clearly illustrate the technical solutions of the embodiments of the present application, the drawings needed to be used in the description of the embodiments of the present application will be briefly introduced. Obviously, the drawings in the following description are only some embodiments of the present application, and other drawings can also be obtained by those skilled in the art without any creative labor.

[0035] Figure 1 Flow chart for the establishment of the benign prostatic hyperplasia model of the rats;

[0036] Figure 2 Statistical chart of the body weight changes of the rats in different administration groups;

[0037] Figure 3 Statistical chart of the prostate weight index and volume index of the rats in different administration groups, wherein A is the prostate weight index and B is the prostate volume index;

[0038] Figure 4 Statistical chart of the prostate lateral lobe index of the rats in different administration groups;

[0039] Figure 5 Statistical chart of the prostate posterior lobe index of the rats in different administration groups;

[0040] Figure 6The statistical chart of testicular index and sperm number of rats in different administration groups, wherein A is testicular index, and B is sperm number;

[0041] Figure 7 The statistical chart of serum T, F-TESTO, DHT and PSA contents of rats in different administration groups, wherein A is T content, B is F-TESTO content, C is DHT content, and D is PSA content;

[0042] Figure 8 The visual chart of prostate morphology of rats after 60 days of treatment in different administration groups;

[0043] Figure 9 The HE staining chart of prostate tissue of rats after 60 days of treatment in different administration groups, wherein (a) is the NC group, (b) is the D group, (c) is the T group, (d) is the ML group, (e) is the MM group, and (f) is the MH group;

[0044] Figure 10 The HE staining chart of testicular tissue of rats after 60 days of treatment in different administration groups, with a scale of 100 μm;

[0045] Figure 11 The SA-β-gal staining chart of prostate tissue of rats after 60 days of treatment in different administration groups;

[0046] Figure 12 The results of AR, Ki67 and PCNA immunohistochemistry of prostate tissue of rats in different administration groups, wherein A is the AR positive cells in prostate tissue shown by IHC analysis, B is the statistical chart of AR positive staining area percentage in prostate tissue, C is the ki67 positive cells in prostate tissue shown by IHC staining, D is the statistical chart of ki67 positive staining area percentage in prostate tissue, E is the PCNA positive cells in prostate tissue shown by immunohistochemical staining, and F is the statistical chart of PCNA positive staining area percentage in prostate tissue. DETAILED DESCRIPTION

[0047] The technical solutions of the present application are further described below through the drawings and examples.

[0048] In order to make the purpose, technical solutions and advantages of the present application more clear, thorough and complete, the technical solutions of the present application are clearly and completely described below through the drawings and examples. The following detailed description is the description of examples, which aims to provide further detailed description of the present application. Unless otherwise specified, all technical terms used in the present application have the same meaning as generally understood by those skilled in the art to which the present application belongs.

[0049] The instruments and reagents used in the examples are obtained through commercial channels.

[0050] Example One

[0051] A method for establishing a rat benign prostatic hyperplasia model, the flow is as shown in Figure 1 The 8-week-old (200g±20g) male Wistar rats were purchased from Beijing Vito Lihua Experimental Animal Technology Co., Ltd., and the production license number was SCXK (Zhejiang) 2020-0002. All animals were bred in the barrier environment of the Experimental Animal Center of Zhejiang Province Institute of Traditional Chinese Medicine, and the license number was SYXK (Zhejiang) 2019-0010. The light / dark cycle was 12h, and the constant temperature (25℃) and humidity (75%) were maintained. After 7 days of adaptive feeding, the Wistar rats were randomly divided into 6 groups, 6 rats in each group. They were normal group (NC), D-galactose group (D), testosterone undecanoate group (T), D-galactose and testosterone undecanoate combined model low-dose group (ML), D-galactose and testosterone undecanoate combined model medium-dose group (MM), and D-galactose and testosterone undecanoate combined model high-dose group (MH). The experimental period was 60 days.

[0052] During the test period, the D (500mg / kg / d), ML (125mg / kg / d), MM (250mg / kg / d), and MH (500mg / kg / d) groups were given subcutaneous injections of D-galactose at the corresponding doses; from the 30th day, the T, ML, MM, and MH groups were given intraperitoneal injections of testosterone undecanoate olive oil solution at a dose of 15mg / kg / 10d. The NC group was injected with the same volume of normal saline and olive oil. This example scheme was approved by the Experimental Animal Welfare Ethics Committee of Zhejiang Province Institute of Traditional Chinese Medicine, and the ethical review approval number was Zhejiang Research Animal Ethics Review No.

[2024] 033.

[0053] Test Example One

[0054] Body weight, organ index, and sperm count determination

[0055] The body weight of the rats was measured once a week. After the end of the test, the rats were anesthetized with 3% isoflurane inhalation, and the prostate and testes were dissected, photographed, and weighed. The prostate volume was determined using a volume measuring instrument. The organ index was calculated by the following formula:

[0056] .

[0057] Sperm parameters were analyzed using the diffusion method to collect sperm in the tail of the epididymis. The dissected rat epididymis was placed in a 2mL 0.9% sodium chloride solution, and the sperm was released by lightly cutting the tail of the epididymis several times with scissors. The sperm was mixed gently, filtered, and 20μL of the liquid was taken to the cell counting plate, and the sperm count was read under a microscope.

[0058] Data were presented as mean ± standard deviation (SD). One-way ANOVA and independent sample t-test were used to analyze the data using SPSS statistics 25.0. p <0.05 was considered to represent a statistically significant difference ).

[0059] The body weight of rats in each group was as shown in Figure 2 The results showed that there was no statistically significant difference in the body weight of rats in each group.

[0060] The prostate weight and volume index of rats in each group were as shown in Figure 3 , where A was the prostate weight and B was the prostate volume index. The results showed that the prostate weight and volume index of the MM and MH groups were significantly higher than those of the NC group.

[0061] The prostate lateral lobe index of rats in each group was as shown in Figure 4 The results showed that there was no significant difference in the prostate lateral lobe index of rats in each group.

[0062] The prostate posterior lobe index of rats in each group was as shown in Figure 5 The results showed that the prostate posterior lobe index of the ML, MM, and MH groups was significantly higher than that of the NC group.

[0063] The testis and sperm count of rats in each group was observed, as shown in Figure 6 , where A was the testis index and B was the sperm count. The results showed that the testis index and sperm count of rats in the D, ML, MM, and MH groups were lower than those in the NC group, with significant differences.

[0064] Test Example Two

[0065] ELISA detection of serum T, F-TESTO, DHT, PSA content

[0066] The concentrations of serum testosterone (T, CSB-E05100r), dihydrotestosterone (DHT, CSB-E07879r), free testosterone (F-TESTO, CSB-E05097r), and prostate specific antigen (PSA, CSB-E08275r) were detected by enzyme-linked immunosorbent assay (ELISA) kit. The Power Wave XS full-wavelength microplate reader (Gene Company Limited) was used to determine the absorbance at the appropriate wavelength according to the ELISA kit instructions.

[0067] The results are shown in Figure 7 , where A was the testosterone T content, B was the free testosterone F-TESTO content, C was the dihydrotestosterone DHT content, and D was the prostate specific antigen PSA content.

[0068] The experimental results showed that, compared with the NC group, the levels of testosterone (T) and dihydrotestosterone (DHT) in the T, ML, MM, and MH groups were significantly increased; the level of free testosterone (F-TESTO) in the serum of rats in the MM group was different from that in the NC group. Serum prostate-specific antigen (PSA) levels were observed to be significantly higher in the T and MH groups compared with the NC group.

[0069] Test Example 3

[0070] HE staining of prostate and testicular tissues

[0071] Sixty days after model establishment, rats were sacrificed, and prostate tissue was collected. The tissue was fixed in 4% paraformaldehyde, slowly decalcified with EDTA, embedded in paraffin, and sectioned. Sections were baked at 60℃ for 30 min, dewaxed in xylene, and gradually hydrated using a gradient of ethanol solutions. Sections were stained sequentially with hematoxylin and eosin, differentiated with differentiation solution, and then blued with 0.5% ammonia. The sections were mounted with neutral resin, observed and photographed under a microscope, and the results were analyzed. Testicular tissue was fixed and immediately embedded in paraffin; subsequent staining and analysis were performed following the same procedures as for prostate tissue.

[0072] like Figure 8 The image shown is a visual representation of the morphology of the rat prostate (the entire gland, with the lateral and posterior lobes of the prostate in the rectangular frame). It can be observed that androgens can induce an increase in prostate volume, and the lateral and posterior lobes show significantly thicker and fuller tissues.

[0073] like Figure 9 The images shown are HE staining images of rat prostate tissue (magnification: ×100 and ×200; scale bar = 250μm & 100μm). (a) represents the NC group, (b) the D group, (c) the T group, (d) the ML group, (e) the MM group, and (f) the MH group. HE staining results show that in the NC group, the prostate tissue of rats is full and regular, with a single layer of columnar cells in the glandular epithelium and loose stroma. After different modeling methods, the rat prostate glands are irregular, with hyperplasia of glandular epithelial cells, varying degrees of single-layer tall columnar cells forming mucosal folds, and inflammatory cell infiltration. The MH group showed the most significant glandular damage.

[0074] Reproductive dysfunction is an important indicator of aging and metabolic abnormalities. Previous experimental data have shown that the testicular index of rats in the D, ML, MM, and MH groups was significantly lower than that in the NC group. Subsequently, pathological sections of the testes of rats in each group were examined, such as... Figure 10 As shown.

[0075] The results showed that the NC group had normal tissue structure, complete spermatogenic epithelium, and orderly arranged spermatogenic cells and Sertoli cells in the basement membrane, and Leydig cells distributed in groups. After modeling in different ways, the seminiferous tubules in the testis tissue of rats atrophied, the spermatogenic epithelium thinned, the spermatogenic cells arranged loosely and disorderly, the Leydig cell gap enlarged, and the number reduced. Among them, the D and MH groups had the most obvious testicular damage.

[0076] Test Example Four

[0077] SA-β-gal staining of prostate tissue

[0078] The prostate tissue block was embedded in glue and frozen in a-20°C freezing microtome. After solidification, the liver tissue block was cut into 5 μm thick sections and adhered to the glass slide patch. The frozen sections were warmed, washed with PBS for 3 times, and then an appropriate volume of SA-β-gal staining fixative (#C0602, Beyotime Biotechnology, Shanghai, China) was added and fixed at room temperature for not less than 15 min. Then the sample was washed with PBS, an appropriate staining working solution was added, and incubated at 37°C overnight, and then observed under a microscope.

[0079] The results are shown in Figure 11 (×200 magnification; scale = 100 μm), and the experimental results showed that compared with the NC group, the D, ML, MM, and MH groups had larger SA-β-gal blue staining areas in the prostate tissue, and more SA-β-gal positive cells, and the blue staining sites were mainly distributed in the glandular epithelial cells, among which the D and MH groups were the most obvious.

[0080] Test Example Five

[0081] Immunohistochemical detection of AR, Ki67, and PCNA protein expression in prostate tissue

[0082] Prostate tissue was paraffin-embedded and sectioned. After dewaxing and antigen retrieval, the paraffin-embedded sections were incubated overnight at 4°C with anti-AR antibody (#22089-1-AP, 1:5000, Proteintech Group, Wuhan, China), anti-Ki67 antibody (#9129, 1:100, Cell Signaling Technology, Boston, USA), or anti-PCNA antibody (#BM0104, 1:2000, Boster Company, USA), respectively. Then, they were incubated at 37°C for 50 minutes with goat anti-rabbit (IgG) secondary antibody HRP (1:2000, #ab205718, Abcam, China). After washing with PBS, the sections were developed using the peroxidase substrate DAB (#DAB-4033, MXBBiotechnologies, Fuzhou, China). After staining, differentiation, and sealing, the sections were observed and photographed under a microscope. ImageJ was used to analyze randomly captured images to determine the optical density of positive targets in different randomly selected fields of view. The expression level of the target indicator is expressed as the percentage of positively stained cells. Representative images are presented to illustrate the findings.

[0083] Immunohistochemical results of AR, Ki67, and PCNA in prostate tissues of rats in each group are as follows: Figure 12 As shown, the magnification is 200x. A shows AR-positive cells in prostate tissue as seen by IHC analysis (magnification: ×200; scale bar = 100μm); B is a statistical graph of the percentage of AR-positive staining area in prostate tissue; C shows Ki67-positive cells in prostate tissue stained with IHC (magnification: ×200; scale bar = 100μm); D is a statistical graph of the percentage of Ki67-positive staining area in prostate tissue; E shows PCNA-positive cells in prostate tissue stained with immunohistochemicals (magnification: ×200; scale bar = 100μm); and F is a statistical graph of the percentage of PCNA-positive staining area in prostate tissue. Compared with the NC group, , .

[0084] The results showed that the expression level of androgen receptor AR was significantly increased in the MM and MH groups, which may be one of the underlying mechanisms for the more stable models in these two groups. In addition, the percentage of Ki67 positive staining area was significantly increased in the MM and MH groups and PCNA positive staining area in the T, MM and MH groups, suggesting that these two genes may be involved in the excessive proliferation of prostate cells.

[0085] It should be pointed out finally that the above examples are only used to illustrate the technical solutions of the present application but not to limit it, and although the present application has been described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that the technical solutions of the present application can still be modified or replaced equivalently, and these modifications or equivalent replacements should not make the modified technical solutions deviate from the spirit and scope of the technical solutions of the present application.

Claims

1. A method for establishing a rat model of benign prostatic hyperplasia, characterized by comprising the following steps: D-galactose and testosterone undecanoate were combinedly administered to the rats under adaptive feeding to induce benign prostatic hyperplasia; ​ The induction time was 60 days, during which 125mg / kg / d-500mg / kg / d of D-galactose was subcutaneously injected, and 15mg / kg / 10d of testosterone undecanoate olive oil solution was intraperitoneally injected from the 30th day.

2. The method for establishing a rat model of benign prostatic hyperplasia according to claim 1, characterized in that: The rats were Wistar rats.

3. The method for establishing a rat model of benign prostatic hyperplasia according to claim 1, characterized in that, The adaptive feeding conditions were as follows: the rats were bred in a barrier environment, with 12h light / dark cycle, temperature of 25℃, humidity of 75%, and adaptive feeding for 7 days.

4. The method for establishing a rat model of benign prostatic hyperplasia according to claim 1, characterized in that: The subcutaneous injection dose of D-galactose was 125mg / kg / d.

5. The method for establishing a rat model of benign prostatic hyperplasia according to claim 1, characterized in that: The subcutaneous injection dose of D-galactose was 250mg / kg / d.

6. The method for establishing a rat model of benign prostatic hyperplasia according to claim 1, characterized in that: The subcutaneous injection dose of D-galactose was 500mg / kg / d.

7. Use of the rat benign prostatic hyperplasia model according to any one of claims 1-6 in the study of the pathogenesis of benign prostatic hyperplasia.

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