Rat benign prostatic hyperplasia model, establishment method and application thereof

Abstract

The invention discloses a rat benign prostatic hyperplasia model, an establishment method and application thereof, and belongs to the technical field of animal models. The establishment method comprises the following steps: jointly applying D-galactose and testosterone undecanoate to adaptively fed rats to induce benign prostatic hyperplasia; the induction time is 60 days, the hypodermic injection of 125mg / kg / d to 500mg / kg / d of D-galactose is continuously carried out within 60 days, and the intraperitoneal injection of 15mg / kg / 10d of testosterone undecanoate olive oil solution is carried out on the 30th day. The effectiveness and practicability of the model are comprehensively evaluated by detecting the prostate index, serum hormone level, histopathology, androgen receptor, proliferation related molecules and testis function of the rat benign prostatic hyperplasia model jointly induced by D-galactose and testosterone undecanoate. The result shows that the aging and androgen combined induced rat prostatic hyperplasia model constructed by the invention is closer to the pathogenesis characteristic of clinical benign prostatic hyperplasia.

Description

Technical Field

[0001] This invention relates to the field of animal model technology, and in particular to a rat benign prostatic hyperplasia model, its establishment method, and its application. Background Technology

[0002] Benign prostatic hyperplasia (BPH) is a common disease in middle-aged and elderly men. It is mainly characterized by histological hyperplasia of the prostatic stroma and glandular components, anatomical enlargement of the prostate, and urodynamic obstruction of the bladder outlet (BOO). Clinical symptoms include storage urinary frequency, urgency, and incontinence; voiding symptoms such as hesitation, difficulty, or intermittent urination; and post-void dribbling, as well as related complications. Most men over 65 years of age have histological findings suggestive of BPH, and about two-thirds of these men will experience clinical symptoms.

[0003] With the deepening of global population aging, the prevalence of benign prostatic hyperplasia (BPH) continues to rise. Currently, treatment primarily involves medication maintenance and transurethral resection of the prostate (TURP), but due to unstable long-term drug efficacy and high surgical risks in elderly patients, there is still no definitively effective treatment. Therefore, accurately elucidating the molecular mechanisms of BPH pathogenesis and exploring preclinical animal models that can accurately evaluate treatment options are indispensable.

[0004] Rat animal models, due to their advantages of pure breed, large quantity, and low cost, have become important vehicles for studying the pathogenesis of benign prostatic hyperplasia (BPH) and evaluating drug efficacy. In recent years, the establishment of rat models of BPH has mainly included the following methods: (1) Testosterone induction: Since prostate growth is mainly regulated by androgens, the most common modeling method is to induce prostate growth with high doses of exogenous androgens, intervening in normal or castrated (orchiectomized) male rats. This type of model can cause hyperplasia of the prostate epithelium and 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 greater degree of glandular and interstitial hyperplasia in histopathology. However, testosterone, as the most abundant and important androgen in human men, is entirely produced by the interstitial cells of the testes. With age, although the weakening of testicular function will lead to a decrease in the level of endogenous testosterone in the body, the prostate is different from other androgen-dependent organs, and its receptors have the ability to respond to androgens throughout life.

[0005] Clinical studies have found that the levels of dihydrotestosterone (DHT), a derivative of testosterone, and its receptors remain high in the prostate of elderly men. The testes continuously supply testosterone to maintain DHT production in the prostate, which is a key driving factor in the development of benign prostatic hyperplasia (BPH). Therefore, castration has certain limitations: firstly, the surgery is difficult, and the glandular tissue is difficult to restore to pre-castration levels after hyperplasia, limiting the assessment of its effectiveness; secondly, it fails to reflect the aging characteristics of clinical BPH and the presence but decline of testicular function, making it difficult to objectively reflect the core pathological changes of BPH.

[0006] (2) Synergistic induction by estrogen and androgen: Studies have shown that with increasing age, the prostate volume gradually increases, and the serum estrogen-androgen ratio shows an upward trend. Therefore, synergistic intervention with estrogen and androgen in castrated or non-castrated rats can exacerbate the imbalance of endogenous estrogen-androgen ratio, leading to benign prostatic hyperplasia. This model has a high degree of consistency with clinical diagnostic criteria, but the estrogen-androgen ratio is often difficult to accurately assess, making it difficult to determine the hormone ratio at the time of model establishment, which to some extent affects the success rate of model establishment.

[0007] (3) Urogenital sinus implantation method: Some researchers have implanted urogenital sinus tissue from fetal mice of the same strain into the ventral lobe of the prostate to stimulate the proliferation of embryonic tissue around the prostate urethra in model animals and induce benign prostatic hyperplasia. However, this method has problems such as high operation difficulty, high risk of infection, and high animal mortality.

[0008] (4) Metabolic syndrome model: Multiple preclinical and clinical studies have shown that metabolic abnormalities are an important determinant of the occurrence and progression of benign prostatic hyperplasia (BPH) / LUTS. Therefore, abnormalities in prostate morphology and function are often observed in animal models of metabolic syndrome, such as high-fat diet and spontaneously hypertensive rats. However, this method is only suitable for exploring the relationship between BPH and metabolic diseases, with a narrow application scenario and a low model success rate.

[0009] Aging and functional testes are recognized as two major factors in the clinical pathogenesis of benign prostatic hyperplasia (BPH). However, a BPH model that addresses both factors is currently lacking. This invention constructs a BPH model induced by the aging inducer D-galactose and testosterone undecanoate. This model introduces aging factors to weaken the endogenous endocrine regulation of the testes and enhance the sensitivity of the prostate to exogenous androgens. The effectiveness and practicality of the model are comprehensively evaluated by monitoring changes in prostate index, serum hormone levels, histopathology, androgen receptors and proliferation-related molecules, as well as testicular function. Summary of the Invention

[0010] The purpose of this invention is to provide a rat model of benign prostatic hyperplasia, its establishment method, and its application, in order to solve the problem that existing models cannot accurately simulate clinical pathogenesis characteristics.

[0011] To achieve the above objectives, the present invention provides a method for establishing a rat model of benign prostatic hyperplasia, wherein benign prostatic hyperplasia is induced by administering the aging inducer D-galactose and the androgen testosterone undecanoate to adaptively fed rats. The induction period was 60 days, during which D-galactose was administered subcutaneously at a dose of 125 mg / kg / day to 500 mg / kg / day. Starting on day 30, testosterone undecanoate solution was administered intraperitoneally at a dose of 15 mg / kg / 10 days.

[0012] The preferred rat is the Wistar rat.

[0013] Preferred adaptive feeding conditions are: raising in a barrier environment with a 12-hour light / dark cycle, a temperature of 25°C, a humidity of 75%, and 7 days of adaptive feeding.

[0014] Preferably, the subcutaneous injection dose of D-galactose is 125 mg / kg / day.

[0015] Preferably, the subcutaneous injection dose of D-galactose is 250 mg / kg / day.

[0016] Preferably, the subcutaneous injection dose of D-galactose is 500 mg / kg / day.

[0017] On the other hand, the present invention provides an evaluation method for the above-mentioned rat benign prostatic hyperplasia model, including at least one of the following evaluation criteria: (1) Physiological indicators: prostate index, testicular index and epididymal tail sperm count; (2) Serum hormone levels: serum testosterone, dihydrotestosterone, free testosterone and prostate-specific antigen concentrations; (3) Histopathology: HE staining was used to observe the structure of the prostate gland and the state of the seminiferous tubules of the testes, and SA-β-gal staining was used to detect senescent cells in the prostate tissue; (4) Molecular markers: Immunohistochemistry was used to detect the percentage of positive cells for androgen receptor, proliferation marker Ki67 and PCNA in prostate tissue.

[0018] On the other hand, the present invention provides an application of the above-mentioned rat benign prostatic hyperplasia model in the study of the pathogenesis of benign prostatic hyperplasia.

[0019] On the other hand, the present invention provides an application of the above-mentioned rat benign prostatic hyperplasia model in the screening and evaluation of drugs for benign prostatic hyperplasia.

[0020] Therefore, the rat benign prostatic hyperplasia model, its establishment method, and its application, as described in this invention, have the following beneficial effects: (1) This invention induces a rat model of benign prostatic hyperplasia by using the aging inducer D-galactose and testosterone undecanoate. The effectiveness and practicality of the model are further evaluated by changes in prostate index, serum hormone levels, histopathology, androgen receptors and proliferation-related molecules and testicular function. The results show that the rat model of benign prostatic hyperplasia induced by aging and androgens constructed in this invention is closer to the pathogenesis of clinical benign prostatic hyperplasia. (2) In the process of evaluating the rat model, the present invention found that the single aging inducer D-galactose could not induce prostatic hyperplasia in rats, and the single androgen testosterone undecanoate induced prostatic hyperplasia model in rats had poor stability. (3) Analysis of the prostate lateral or posterior lobe index of rats revealed no difference in the prostate lateral lobe of rats in different drug administration groups or different dose groups. The prostate posterior lobe index of rats in the D-galactose and testosterone undecanoate combined administration group was significantly increased compared with the blank treatment group, which is more similar to the peripheral area structure of humans that is prone to disease. (4) The serum levels of different forms of testosterone in each group of rats were detected. The results showed that the abundance of T, F-TESTO and DHT in rats treated with testosterone undecanoate increased to varying degrees. The prostate-specific antigen (PSA) level was significantly increased in 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 showed that after the establishment of the D-galactose and testosterone undecanoate combined model, the folds of the prostate gland mucosa of rats were prominent, the glandular epithelial cells were significantly hyperplastic and inflammatory infiltrated, and a lot of SA-β-Gal positive staining was observed. It can be seen that the rat benign prostatic hyperplasia model induced by D-galactose and testosterone undecanoate is more consistent with the pathogenesis of clinical benign prostatic hyperplasia and has a better modeling effect. (5) The levels of AR, Ki67, and PCNA in rat prostate tissue were detected by immunohistochemistry. The results showed that after modeling, AR was significantly increased in the medium-dose group and the high-dose group of the D-galactose and testosterone undecanoate combined model. At the same time, compared with the normal group, the percentage of Ki67 and PCNA positive cells in the medium-dose group and the high-dose group of the D-galactose and testosterone undecanoate combined model was significantly increased. This suggests that the rat benign prostatic hyperplasia model induced by D-galactose and testosterone undecanoate can cause excessive proliferation of prostate cells through proteins such as Ki67 and PCNA. (6) Calculations of testicular index and sperm count revealed that the testicular index and sperm count of rats treated with the aging-inducing drug D-galactose were significantly reduced. Furthermore, histopathological examination directly showed that aging can cause damage to testicular tissue cells. This suggests that aging and androgen stimulation can jointly cause damage to the testicular structure in rats.

[0021] The technical solution of the present invention will be further described in detail below with reference to the accompanying drawings and embodiments. Attached Figure Description

[0022] To more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings used in the description of the embodiments of the present invention will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0023] Figure 1 Flowchart for establishing a rat model of benign prostatic hyperplasia; Figure 2 A statistical graph showing the changes in body weight of rats in different drug administration groups; Figure 3 The graph shows the prostate weight index and volume index of rats in different drug administration groups. In the graph, A is the prostate weight index and B is the prostate volume index. Figure 4 Statistical graph of prostate lateral lobe index in rats of different drug administration groups; Figure 5 Statistical graph of the posterior prostate index in rats of different drug administration groups; Figure 6 The graph shows the testicular index and sperm count of rats in different drug administration groups. In the graph, A represents the testicular index and B represents the sperm count. Figure 7 The graph shows the serum levels of T, F-TESTO, DHT, and PSA in rats from different drug administration groups. In the graph, A represents the T level, B represents the F-TESTO level, C represents the DHT level, and D represents the PSA level. Figure 8 Visual representation of prostate morphology in rats from different drug treatment groups after 60 days of treatment; Figure 9 HE staining images of prostate tissue from rats in different drug treatment groups after 60 days of treatment. (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. Figure 10 HE staining images of testicular tissue from rats in different drug treatment groups after 60 days of treatment. Scale bar: 100 μm. Figure 11SA-β-gal staining images of prostate tissues from rats in different administration groups after 60 days of treatment; Figure 12 Immunohistochemical results of AR, Ki67, and PCNA in prostate tissues of rats in different administration groups. Among them, A shows AR-positive cells in prostate tissues by IHC analysis, B is the statistical chart of the percentage of AR-positive staining area in prostate tissues, C shows Ki67-positive cells in prostate tissues by IHC staining, D is the statistical chart of the percentage of Ki67-positive staining area in prostate tissues, E shows PCNA-positive cells in prostate tissues by immunohistochemical staining, and F is the statistical chart of the percentage of PCNA-positive staining area in prostate tissues. Detailed implementation manner

[0024] The technical solutions of the present invention will be further described below with reference to the accompanying drawings and examples.

[0025] In order to make the purpose, technical solutions and advantages of this application clearer, more thorough and complete, the technical solutions of the present invention will be clearly and completely described below with reference to the accompanying drawings and examples. The following detailed descriptions are all descriptions of examples, aiming to provide further detailed descriptions of the present invention. Unless otherwise specified, all technical terms used in the present invention have the same meaning as commonly understood by those of ordinary skill in the art to which this application belongs.

[0026] The instruments, equipment, reagents and materials used in the examples were all obtained through commercial channels.

[0027] Example 1 A method for establishing a rat model of benign prostatic hyperplasia, the process is as Figure 1 shown, including: Male Wistar rats at 8 weeks old (200 g ± 20 g) were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd., production license number: SCXK (Zhe) 2020-0002. All animals were housed in the barrier environment of the Experimental Animal Center of Zhejiang Academy of Traditional Chinese Medicine, license number: SYXK (Zhe) 2019-0010. The light / dark cycle was 12 h, with a constant temperature (25 °C) and humidity (75%). After 7 days of adaptive feeding, the Wistar rats were randomly divided into 6 groups, with 6 rats in each group. They were the normal group (NC), D-galactose group (D), testosterone undecanoate group (T), low-dose combined model group of D-galactose and testosterone undecanoate (ML), medium-dose combined model group of D-galactose and testosterone undecanoate (MM), and high-dose combined model group of D-galactose and testosterone undecanoate (MH), and the experimental period was 60 d.

[0028] During the experiment, groups D (500 mg / kg / d), ML (125 mg / kg / d), MM (250 mg / kg / d), and MH (500 mg / kg / d) received subcutaneous injections of the corresponding doses of D-galactose. From day 30 onwards, groups T, ML, MM, and MH received intraperitoneal injections of testosterone undecanoate in olive oil solution at a dose of 15 mg / kg / 10 days. Group NC received an equal volume of physiological saline and olive oil. This protocol was approved by the Experimental Animal Welfare and Ethics Committee of Zhejiang Academy of Traditional Chinese Medicine, with ethics review approval number: Zhezhongyan Animal Ethics Review No.

[2024] 033.

[0029] Test Example 1 Measurement of body weight, organ index and sperm count Rats were weighed weekly. After the experiment, rats were anesthetized with 3% isoflurane inhalation, and the prostate and testes were dissected, photographed, and weighed. Prostate volume was measured using a volumetric analyzer. Organ index was calculated using the following formula: .

[0030] Sperm were collected from the epididymal tail using a diffusion method for sperm parameter analysis. The rat epididymis was dissected and placed in 2 mL of 0.9% sodium chloride solution. The epididymal tail was gently cut several times with scissors to release the sperm. After gentle mixing, 20 μL of the liquid was collected on a cell counting plate and the sperm count was read under a microscope.

[0031] Data are presented as mean ± standard deviation (SD). One-way ANOVA and independent samples t-tests were performed using SPSS Statistics 25.0 to analyze the data. p <0.05 is considered to represent a statistically significant difference ( ).

[0032] The body weights of rats in each group are as follows: Figure 2 As shown in the figure, the results indicated no statistically significant difference in the body weight of rats in each group.

[0033] The prostate weight and volume index of rats in each group are as follows: Figure 3 As shown in the figure, A represents prostate weight and B represents prostate volume index. The results showed that, compared with the NC group, the prostate weight and volume index were significantly higher in the MM and MH groups.

[0034] The prostate lateral lobe index of each group of rats is as follows: Figure 4 As shown in the figure, the results indicated no significant differences in the prostate lateral lobe index among the groups of rats.

[0035] The posterior prostate index of each group of rats is as follows: Figure 5 As shown in the results, the posterior prostate index of the ML, MM, and MH groups was significantly higher than that of the NC group.

[0036] Observe the testes and sperm count of rats in each group, such as Figure 6 As shown in the figure, A represents the testicular index and B represents the sperm count. The results showed that the testicular index and sperm count of rats in groups D, ML, MM, and MH were significantly lower than those in group NC.

[0037] Test Example 2 ELISA detection of serum T, F-TESTO, DHT, and PSA levels Serum testosterone (T, CSB-E05100r), dihydrotestosterone (DHT, CSB-E07879r), free testosterone (F-TESTO, CSB-E05097r), and prostate-specific antigen (PSA, CSB-E08275r) concentrations were detected using an enzyme-linked immunosorbent assay (ELISA) kit. Absorbance measurements were performed using a Power Wave XS full-wavelength ELISA reader (Gene Company Limited), with appropriate wavelengths set according to the ELISA kit instructions.

[0038] The results are as follows Figure 7 As shown, A represents testosterone (T) content, B represents free testosterone (F-TESTO) content, C represents dihydrotestosterone (DHT) content, and D represents prostate-specific antigen (PSA) content.

[0039] 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.

[0040] Test Example 3 HE staining of prostate and testicular tissues 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.

[0041] 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.

[0042] 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.

[0043] 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.

[0044] The results showed that the NC group had normal tissue structure, intact seminiferous epithelium, and neatly arranged spermatocytes and Sertoli cells of various grades within the basement membrane, with testicular interstitial cells distributed in clusters. After different modeling methods, the seminiferous tubules in the rat testicular tissue atrophied, the seminiferous epithelium thinned, the spermatogenic cells were loosely and disordered, and the interstitial cells of the testis had enlarged gaps and reduced numbers. The testicular damage was most obvious in the D and MH groups.

[0045] Test Example 4 SA-β-gal staining of prostate tissue Prostate tissue blocks were embedded in adhesive and frozen at -20°C using a cryostat. After solidification, liver tissue blocks were cut into 5μm thick sections and attached to glass slides. Frozen sections were thawed, washed three times with PBS, and fixed at room temperature for at least 15 minutes with an appropriate volume of SA-β-gal staining fixative (#C0602, Beyotime Biotechnology, Shanghai, China). The samples were then washed with PBS, and an appropriate amount of staining working solution was added. The samples were incubated overnight at 37°C and then observed under a microscope.

[0046] The results are as follows Figure 11 As shown (magnification: ×200; scale bar = 100μm), the experimental results show that, compared with the NC group, the prostate tissues of rats in the D, ML, MM, and MH aging model groups had a larger area of ​​SA-β-gal blue staining and more SA-β-gal positive cells. The blue staining sites were mainly distributed in glandular epithelial cells, with the D and MH groups showing the most obvious results.

[0047] Test Example 5 Immunohistochemistry was used to detect the expression levels of AR, Ki67, and PCNA proteins in prostate tissue. 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.

[0048] 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, , .

[0049] 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.

[0050] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and not to limit them. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can still be made to the technical solutions of the present invention, and these modifications or equivalent substitutions cannot cause the modified technical solutions to deviate from the spirit and scope of the technical solutions of the present invention.

Claims

1. A method for establishing a rat model of benign prostatic hyperplasia, characterized in that: Benign prostatic hyperplasia was induced in adaptively fed rats by combined administration of D-galactose and testosterone undecanoate. The induction period was 60 days, during which D-galactose was administered subcutaneously at a dose of 125 mg / kg / day to 500 mg / kg / day. Starting on day 30, testosterone undecanoate solution was administered intraperitoneally at a dose of 15 mg / kg / 10 days.

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, Adaptation feeding conditions were as follows: the animals were kept in a barrier environment with a 12-hour light / dark cycle, a temperature of 25°C, and a humidity of 75% 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 is 125 mg / kg / day.

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 is 250 mg / kg / day.

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 is 500 mg / kg / day.

7. A rat benign prostatic hyperplasia model established by the method for establishing a rat benign prostatic hyperplasia model as described in any one of claims 1 to 6.

8. An evaluation method for a rat benign prostatic hyperplasia model as described in claim 7, characterized in that, Include at least one of the following evaluation criteria: (1) Physiological indicators: prostate index, testicular index and epididymal tail sperm count; (2) Serum hormone levels: serum testosterone, dihydrotestosterone, free testosterone and prostate-specific antigen concentrations; (3) Histopathology: HE staining was used to observe the structure of the prostate gland and the state of the seminiferous tubules of the testes, and SA-β-gal staining was used to detect senescent cells in the prostate tissue; (4) Molecular markers: Immunohistochemistry was used to detect the percentage of positive cells for androgen receptor, proliferation marker Ki67 and PCNA in prostate tissue.

9. The application of the rat benign prostatic hyperplasia model as described in claim 7 in the study of the pathogenesis of benign prostatic hyperplasia.

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