Application of pawpaw total triterpenes in preparation of medicine for treating benign prostatic hyperplasia

Abstract

The invention provides a 5 alpha-reductase inhibitor. The inhibitor is pawpaw total triterpenes (TCS). The invention also provides application of the pawpaw total triterpenes TCS in preparation of drugs for treating benign prostatic hyperplasia. TCS and UA have no obvious toxic and side effects on visceral organs of each group of rats; the thickness of glandular epithelial cells in a TCS group is remarkably reduced, the 5AR activity of the TCS group is remarkably reduced, the T level is remarkably reduced, and the DTH level is remarkably reduced; the blood sugar of the TCS group is obviously reduced, the concentration of total cholesterol and low-density lipoprotein is obviously reduced, and the TCS can obviously increase the high-density lipoprotein level and reduce the content of triglyceride. The results show that the TCS and the main component tartaric acid have a remarkable inhibiting effect on prostatic hyperplasia. The inhibition of 5AR activity and the reduction of DHT generation and T content are one of the action mechanisms, and the important characteristic is that the prostatic hyperplasia can be inhibited and abnormal blood sugar and blood fat can be regulated at the same time.

Description

Technical Field

[0001] The present invention relates to a pharmaceutical use of total triterpenoids from Chaenomeles sinensis, and particularly relates to a therapeutic use for treating benign prostatic hyperplasia by inhibiting 5α-reductase. Background Art

[0002] Benign Prostatic Hyperplasia (BPH) is a common urological disease, which mostly occurs in middle-aged and elderly men. Epidemiological reports show that among men aged 50 and above, the prevalence of BPH is 50% - 75%; when the age reaches 70 and above, the prevalence of this disease is as high as 80%. The prevalence of BPH shows a trend of gradually increasing with age, and the prevalence of BPH in men under 40 years old is also increasing year by year. Currently, it is generally believed that benign prostatic hyperplasia is mainly related to sex hormone imbalance. In the prostate tissue, testosterone (T) is converted under the catalytic action of 5α-reductase (5AR) to generate dihydrotestosterone (DHT). The binding ability of DHT to androgen receptors is 3 - 10 times higher than that of T. When the activity of 5α-reductase increases, a large amount of DHT is generated, promoting the over-differentiation and proliferation of prostate stromal cells and epithelial cells, and inducing BPH. 5α-Reductase Inhibitors are first-line drugs for treating BPH clinically, but it is found in clinical use that these drugs will produce adverse reactions such as low libido, erectile dysfunction, and reduced ejaculate volume. Therefore, finding safe and effective drugs has become a research hotspot for the treatment of BPH. Traditional Chinese medicine has advantages such as definite curative effect and few adverse reactions in the treatment of benign prostatic hyperplasia. There is a good prospect for finding drugs for treating benign prostatic hyperplasia from traditional Chinese medicine.

[0003] Chaenomeles sinensis (Thouin) Koehne var. wilsonii (Rehd.) Hort. Chaenomeles speciosa (Sweet) Nakai] is the dried nearly mature fruit of the plant of the Rosaceae family, and contains various chemical components such as terpenoids, flavonoids, organic acids, phenols, and polysaccharides. Among them, terpenoid compounds are one of the most important chemical components for the pharmacological action of C. speciosa, and are mainly pentacyclic triterpenoids, including ursolic acid, oleanolic acid, maslinic acid, betulinic acid, etc. However, there is no research report on the effect of total triterpenoids from C. sinensis and its active components on 5α-reductase. The present invention aims to explore the effect of total triterpenoids from C. sinensis on 5α-reductase, so as to provide a basis for further using C. sinensis to treat BPH. Summary of the Invention

[0004] In the present invention, a benign prostatic hyperplasia (BPH) animal model was constructed. Total triterpenoids of Chaenomeles speciosa (TCS) were extracted from Chaenomeles speciosa as raw material. By inhibiting 5AR activity and through the therapeutic effect on BPH rats, the effects of TCS and its main component ursolic acid (UA) were observed. The experimental results showed that TCS and UA could significantly inhibit the 5AR activity in the serum of BPH model rats, effectively reduce the increase in serum T level of BPH rats caused by exogenous testosterone propionate (TP), and further reduce the production of DHT. The results of blood glucose and lipid detection showed that BPH rats induced by testosterone propionate might induce hyperglycemia and hyperlipidemia, which might be the result of the imbalance between male and female hormones. The present invention showed that finasteride could promote the aggravation of abnormal blood lipids, while TCS and UA in different dose groups could significantly improve the abnormal blood glucose and lipid metabolism associated with BPH model rats, specifically manifested as significantly reducing the levels of TC, TG and LDL, and simultaneously increasing the level of HDL, indicating that it had a significant regulatory effect on blood glucose and lipid metabolism disorders. It was suggested that while improving the pathological state of BPH, TCS could also regulate the metabolic status of patients, reduce the occurrence of complications, and had important significance for the prevention and treatment of metabolic syndrome associated with BPH patients. At the same time, Chaenomeles speciosa also had the function of protecting the liver, and this hepatoprotective effect could improve the abnormal blood glucose and lipids caused by the decline of liver function, indicating that TCS had more extensive pharmacological activities than the positive drug finasteride.

[0005] Based on the above content, the present invention provides a 5α-reductase inhibitor, and the inhibitor is total triterpenoids of Chaenomeles speciosa TCS.

[0006] The preparation method of the total triterpenoids of Chaenomeles speciosa TCS includes the following steps: After Fructus Chaenomelis was crushed, it was extracted with 60-95% ethanol as the solvent, concentrated, extracted with petroleum ether and ethyl acetate, the ethyl acetate extraction part was adsorbed by D101 macroporous adsorption resin, and then eluted and separated with ethanol of different concentrations. After the 95% ethanol eluate was recovered from ethanol and concentrated and dried, total triterpenoids of Chaenomeles speciosa were obtained, with a purity of 68.89%, and ursolic acid and oleanolic acid as the main components.

[0007] The application of the 5α-reductase inhibitor in the preparation of a drug for treating prostatic hyperplasia.

[0008] The present invention provides an application of total triterpenoids of Chaenomeles speciosa TCS in the preparation of a drug for treating prostatic hyperplasia.

[0009] The application of the total triterpenoids of Chaenomeles speciosa TCS in the preparation of a drug for treating prostatic hyperplasia by achieving the reduction of the total volume and wet weight of the prostate and seminal vesicles.

[0010] The application of the total triterpenoids of Chaenomeles speciosa TCS in the preparation of a drug for treating prostatic hyperplasia by improving cell morphology, making the vesicular tubular structure clearly visible, reducing the folds in the acinus, and reducing the thickness of the glandular epithelium.

[0011] Use of the total triterpenoids of papaya TCS in the preparation of a drug for treating benign prostatic hyperplasia by reducing the elevated serum T level caused by exogenous testosterone propionate TP.

[0012] Use of the total triterpenoids of papaya TCS in the preparation of a drug for treating benign prostatic hyperplasia by reducing the serum DHT level in BPH.

[0013] Use of the total triterpenoids of papaya TCS in the preparation of a drug for treating benign prostatic hyperplasia by reducing blood glucose.

[0014] Use of the total triterpenoids of papaya TCS in the preparation of a drug for treating benign prostatic hyperplasia by reducing blood lipid.

[0015] For the above uses, the dosage of the total triterpenoids of papaya TCS is 0.5 - 2.0 g·kg -1 。

[0016] This study explored the therapeutic effects and mechanisms of TCS and its main component UA on BPH. It was found that TCS and UA have significant therapeutic effects on BPH, can improve the morphological and pathological states of the prostate tissue, regulate hormone levels and blood glucose and lipid metabolism, and its mechanism of action is related to inhibiting 5AR activity and regulating abnormal lipid metabolism. The present invention helps to further expand the clinical application value of papaya. The current experimental research scope is only limited to the animal model level, lacking in-depth research on cell signaling pathways and molecular levels and verification of clinical trials; although the effects of TCS and UA on 5AR, hormone levels, lipid metabolism and prostate tissue pathology were explored in this study, the specific mechanism of action has not been fully clarified. Brief Description of the Drawings

[0017] Figure 1 Inhibitory effect of the main components of papaya on 5α-reductase.

[0018] Figure 2 Effect of the main components of papaya on the 24 h survival rate of BPH-1 cells.

[0019] Figure 3 Effect of the main components of papaya on the 48 h survival rate of BPH-1 cells.

[0020] Figure 4 Weight changes of BPH rats during the modeling and drug administration period.

[0021] Figure 5 Organ coefficient.

[0022] Figure 6 Effect of TCS on the sex organs of BPH rats.

[0023] Figure 7Changes in wet weight and prostate index of rats. Note: Compared with the Control group, * P < 0.05, ** P < 0.01; compared with the Model group, # P < 0.05, ## P < 0.01.

[0024] Figure 8 HE staining of rat prostate.

[0025] Figure 9 Thickness and area ratio of rat prostate epithelial cells. Note: Compared with the Control group, * P < 0.05, ** P < 0.01; compared with the Model group, # P < 0.05, ## P < 0.01.

[0026] Figure 10 5AR standard curve and the effect of TCS on 5AR level in BPH rats. Note: Compared with the Control group, * P < 0.05, ** P < 0.01; compared with the Model group, # P < 0.05, ## P < 0.01.

[0027] Figure 11 T standard curve and the effect of TCS on T level in BPH rats. Note: Compared with the Control group, * P < 0.05, ** P < 0.01; compared with the Model group, # P < 0.05, ## P < 0.01.

[0028] Figure 12 DHT standard curve and the effect of TCS on DHT level in BPH rats. Note: Compared with the Control group, * P < 0.05, ** P< 0.01; compared with the Model group, # P < 0.05, ## P < 0.01.

[0029] Figure 13 Effects of TCS on the serum blood glucose level of BPH rats. Note: Compared with the Control group, * P < 0.05, ** P < 0.01; compared with the Model group, # P < 0.05, ## P < 0.01.

[0030] Figure 14 Changes in the serum lipid content of rats. Note: Compared with the Control group, * P < 0.05, ** P < 0.01; compared with the Model group, # P < 0.05, ## P < 0.01. Specific implementation manners

[0031] Example 1 The stock solutions of ursolic acid, oleanolic acid, chlorogenic acid and protocatechuic acid were respectively diluted with absolute ethanol to final concentrations of 20 μg / mL and 100 μg / mL, and then the optimized 5α-reductase reaction system was used for screening active ingredients in vitro.

[0032] BPH-1 cells in the logarithmic growth phase were inoculated into 96-well plates at a concentration of 1×104 cells / mL, 100 μL per well. The outermost row was sealed with PBS solution and incubated overnight in an incubator at 37 °C and 5% CO2 saturated humidity. After the cells adhered, 100 μL of ursolic acid, oleanolic acid, protocatechuic acid and chlorogenic acid at concentrations of 10, 60, 120 μg / mL were added respectively. The blank control group was added with an equal amount of RPMI 1640 complete medium, and each group was set with 3 replicates. After culturing for 24 and 48 h respectively, 20 μL of MTT (5 mg / mL) was added to each well, and then incubated in the incubator at 37 °C and 5% CO2 saturated humidity in the dark for 4 h. Then, the culture medium was discarded, 150 μL of DMSO was added to each well, and shaken at low speed for 5 min to promote crystal dissolution. The absorbance value (OD value) of each well was measured at a wavelength of 492 nm by an enzyme-linked immunosorbent assay (ELISA) reader, and the cell survival rate was calculated according to the formula. The experiment was repeated in parallel 3 times.

[0033] Cell survival rate (%) =

[0034] Figure 1 It can be seen that the ingredients contained in papaya have an inhibitory effect on 5α-reductase, among which the triterpenoid ingredients ursolic acid and oleanolic acid have a concentration-dependent inhibition on 5α-reductase, and ursolic acid and protocatechuic acid have a stronger inhibitory effect on 5α-reductase.

[0035] Depend on Figure 2 and Figure 3 It can be seen that ursolic acid in papaya has the most obvious inhibitory effect on BPH-1 cells, followed by high concentration (120 μg / mL) of oleanolic acid, while protocatechuic acid and chlorogenic acid have almost no inhibitory effect on the proliferation of BPH-1 cells. Chlorogenic acid tends to promote the proliferation of BPH-1 cells with increasing concentration.

[0036] Example 2 Papaya: collected from Langping Town, Changyang Tujia Autonomous County, Hubei Province, identified by Professor Wang Yubing of Three Gorges University as Malus sphenanthera of the Rosaceae family. Chaenomeles speciosa (Sweet) Nakai] nearly mature fruit; D101 macroporous adsorption resin: Donghong Chemical Co., Ltd.; testosterone propionate (LOT: X14010E99914), ursolic acid (LOT: C15O11Q126879): Shanghai Yuanye Biotechnology Co., Ltd.; olive oil (pharmaceutical grade, LOT: C17295559): Shanghai McLean Biochemical Technology Co., Ltd.; finasteride (LOT: RH531198): Shanghai Yien Chemical Technology Co., Ltd.; sodium carboxymethyl cellulose (LOT: 20141209): Sinopharm Chemical Reagent Co., Ltd.; sodium penicillin (LOT: 20230901): Shaanxi Lixiang Animal Pharmaceutical Co., Ltd.; DHT ELISA Kit (LO: WV05ZR840158), Rat T ELISA Kit (LOT: WV062T602034): Wuhan Yilai Ruite Biotechnology Co., Ltd.; Rat 5AR ELISA Kit (LOT: 111935006207391119): Shanghai Jianglai Biotechnology Co., Ltd.; Triglyceride Test Kit (LOT: 20241211), Total Cholesterol Kit (LOT: 20241217), Glucose Kit (LOT: 20241113), Low-density lipoprotein Kit (LOT: 20241217), High-density lipoprotein Kit (LOT: 20241217): Nanjing Jiancheng Bioengineering Institute.

[0037] Fifty-six SPF-grade male SD rats, weighing (210±10) g, were provided by the Animal Experiment Center of China Three Gorges University and housed in the SPF-grade animal laboratory of the Animal Experiment Center. The feeding conditions were as follows: relative humidity (55±5)%, temperature 20-25 °C, and 12 / 12 h light-dark cycle with free access to food and water. The production license number of experimental animals was SCXK(E)2022-0012, the license number for the use of experimental animal facilities was SYXK(E)2022-0061, and the approval number for animal ethics review was 2024020V1.

[0038] Preparation of total triterpenoids from Chaenomeles speciosa A certain amount of Chaenomeles speciosa was mechanically pulverized and sieved through a 60-mesh sieve to obtain fine powder of Chaenomeles speciosa. An appropriate amount of the fine powder of Chaenomeles speciosa was precisely weighed. The ultrasonic-assisted extraction technique was adopted, and the extraction process was as follows: ultrasonic time 47 min, ethanol concentration 64%, extraction times 3 times, and solid-liquid ratio 1:40. After the extraction was completed, suction filtration was carried out using a suction filtration device, and the extract was concentrated using a vacuum concentration device to obtain a concentrated solution of Chaenomeles speciosa. The above concentrated solution was suspended in double-distilled water and extracted successively with petroleum ether and ethyl acetate. The ethyl acetate layer extract was again concentrated under reduced pressure to obtain a crude extract of total triterpenoids from Chaenomeles speciosa. D-101 macroporous adsorption resin was taken, soaked in 1.5 times the volume of 95% ethanol for 24 hours, and then rinsed with distilled water until there was no ethanol residue, and then loaded onto the column. The crude extract of total triterpenoids from Chaenomeles speciosa was dissolved in double-distilled water and loaded onto the column for adsorption by wet method at a concentration of 2 mg / mL. Water, 20%, 40%, 60%, 80%, and 95% ethanol were used as eluents for elution and separation, and the eluates of each part were collected, and the total triterpenoid content was determined by the vanillin-glacial acetic acid method. The 95% ethanol eluate was collected, concentrated by vacuum evaporation, and the total triterpenoids from Chaenomeles speciosa were obtained. After vacuum freeze-drying, the purity was determined by the vanillin-glacial acetic acid method to be 65.28%, and its main components were ursolic acid and oleanolic acid. The obtained total triterpenoids from Chaenomeles speciosa (TCS) were used for subsequent animal experimental studies.

[0039] Establishment of BPH model and grouping for drug administration Forty-two male SD rats, weighing 200-220 g, were taken. After all the rats were anesthetized with 2% sodium pentobarbital, the rats in the sham operation group were routinely disinfected and the skin was incised to expose the testicles to the air for several seconds without excision, and then immediately sutured; the skin of the remaining rats was routinely disinfected, and both testicles were removed through the scrotum, and the stump was ligated. After ensuring hemostasis, the skin was sutured. The postoperative rats were intramuscularly injected with sodium penicillin (200,000 U / kg / d) for 5 consecutive days to prevent infection. After one week of recovery, the castrated rats were grouped using the random number table method after weighing, divided into 7 groups, with 6 rats in each group, namely the control group (Control group), the model group (Model group), the finasteride group (Fina group, 0.005 g·kg -1), low-dose total triterpenoids of Chaenomeles speciosa group (TCS-L, 0.5 g·kg -1 ), medium-dose total triterpenoids of Chaenomeles speciosa group (TCS-M, 1.0 g·kg -1 ), high-dose total triterpenoids of Chaenomeles speciosa group (TCS-H, 2.0 g·kg -1 ), ursolic acid group (UA, 0.005 g·kg -1 ). Castrated rats were subcutaneously injected with testosterone propionate (TP) at a dose of 5 mg / kg (dissolved in olive oil) every day. The gavage volume was 0.1 mL per 100 g. The Control group was subcutaneously injected with an equal dose of olive oil and gavaged with drugs at the same time. The gavage volume was 1 mL per 100 g. The Control group and the Model group were gavaged with 0.5% sodium carboxymethylcellulose, and the remaining groups were gavaged with drugs at the corresponding concentrations. The model was established and the drugs were administered continuously for 28 days.

[0040] Animal treatment and sample collection After the last administration, the rats were fasted but allowed to drink water for 12 h, weighed, anesthetized by intraperitoneal injection of 2% sodium pentobarbital, and blood was collected from the abdominal aorta. After standing at room temperature for 2 h, the blood was centrifuged at 4000 rpm for 15 minutes at 4℃, and the supernatant was taken to obtain rat serum, which was aliquoted and stored in a -80℃ refrigerator for subsequent biochemical index detection.

[0041] After the rat blood was collected, the prostate and seminal vesicle tissues and other visceral tissues of the rats were separated, washed with PBS to remove blood, and blotted dry with filter paper and weighed.

[0042] Index observation and detection Changes in rat body weight The activity of rats in each group was observed and recorded every day during the drug administration period, and the body weight was recorded every 3 days after the operation.

[0043] Observation of prostate tissue morphology The washed prostate and seminal vesicle tissues were placed on graph paper to observe the tissue volume.

[0044] Calculation of organ coefficient The intact heart, liver, spleen, lung, and kidney were taken, blotted dry with filter paper and weighed, and the organ coefficient was calculated: organ coefficient = organ mass (g) / body mass (g) × 100%.

[0045] Determination of prostate wet weight and prostate index The prostate was separated from the tissue, weighed, and the prostate index was calculated: prostate index (PI) = prostate wet weight (mg) / body mass (g).

[0046] Pathological observation of prostate tissue The prostate tissue was fixed in 4% paraformaldehyde for 24 h, dehydrated, paraffin-embedded, sectioned, and then subjected to HE staining. The pathological changes of the rat prostate tissue were observed under an inverted microscope.

[0047] Determination of the levels of hormones T and DHT in serum Both the T kit and the DHT kit used the competitive ELISA method. The operation was carried out strictly in accordance with the kit instructions. The OD value was measured at a wavelength of 450 nm using an enzyme-linked immunosorbent assay reader, and the concentrations of T and DHT in the rat serum were calculated by plotting a standard curve. Determination of the content of 5AR in serum The 5AR kit used the double antibody sandwich enzyme-linked immunosorbent assay (ELISA). The absorbance (OD value) was measured at a wavelength of 450 nm using an enzyme-linked immunosorbent assay reader, and the concentration of 5AR in the rat serum was calculated by plotting a standard curve.

[0048] Determination of the blood glucose level in serum The change in the content of blood glucose in the rat serum was measured using the glucose oxidase method strictly in accordance with the kit instructions.

[0049] Determination of the blood lipid level in serum Total cholesterol (TC), triglyceride (TG), low-density lipoprotein (LDL), and high-density lipoprotein (HDL) were measured strictly in accordance with the kit instructions.

[0050] Statistical analysis SPSS 25.0 was used for statistical analysis, and GraphPad Prism 8.0.1 was used for graphing. All data of each group were expressed as One-way ANOVA with Dunnett's post hoc test was used for comparison between groups, and P＜ 0.05 was considered statistically significant.

[0051] Effect of TCS on the behavioral activities of rats with benign prostatic hyperplasia No animal deaths were observed during the castration surgery and the period of modeling and drug administration. During the period of modeling and drug administration, the behaviors of rats in each group, such as activity, diet, and defecation, were normal. The body weights of the rats were recorded after the surgery. It can be seen from Figure 4 that there was no significant difference in the body weight of the rats during the drug administration period compared with the control group.

[0052] Effect of TCS on the organ coefficients of rats with benign prostatic hyperplasia The organ indices of rats in each group were statistically analyzed. As shown in Figure 5As shown. The results showed that there were no significant differences in the organ coefficients of each organ between the Model group and the Control group. Compared with the Model group and the Control group, there were also no significant differences in the organ coefficients of the drug intervention group, indicating that administration of TCS at the current dose for 28 days had no obvious toxic and side effects on the organs of rats.

[0053] Effect of TCS on the morphological changes of prostate tissue Observation of the completely dissected prostate found that (as Figure 6 shown), compared with the Control group, the rats in the Model group showed significant prostate enlargement, and the total volume of the prostate and seminal vesicles increased significantly. By comparison, compared with the model group, the prostate volumes of the finasteride group, the TCS-L, TCS-M, TCS-H dose groups, and the UA group all showed a certain degree of reduction.

[0054] Determination of prostate wet weight and prostate index (PI) The results of prostate index are shown in Figure 7 . Compared with the Control group, the prostate wet weights of the rats in the Model group and the drug intervention group increased significantly, and the prostate indices increased significantly, and there were significant differences ( P <0.05), indicating that subcutaneous injection of testosterone propionate for 28 days could successfully induce a BPH model; compared with the Model group, finasteride, medium and high-dose TCS, and UA could significantly reduce the prostate wet weight and prostate index of BPH rats ( P <0.05, P <0.01), and TCS showed a dose-dependent effect. The above results indicate that both TCS and the main active ingredient ursolic acid in papaya have the potential to treat benign prostatic hyperplasia.

[0055] Effect of TCS on the pathological changes of prostate tissue Figure 8 From top to bottom in the figure are the HE tissue section images magnified 40 times, 200 times, and 400 times respectively under the same microscope field of view. HE staining showed that the vesicular tubular structure of the prostate tissue in the Control group was clear, the glandular epithelial cells were arranged neatly, and there was no hyperplasia in the glands and stroma, and no obvious internal folds protruded into the lumen. Compared with the Control group, the epithelial cells in the Model group were arranged disorderly, the number of folds in the alveoli increased significantly, the prostatic columnar epithelial cells showed obvious hyperplasia, and the internal folds increased and protruded into the lumen in a papillary shape.

[0056] The thickness of the glandular epithelial cells in the prostate tissue under a 200-fold field of view was measured using Image j analysis software, and the area ratio of the hyperplastic glandular epithelial cells in the current field of view was calculated to more clearly reflect the degree of hyperplasia of the prostate tissue. The results showed ( Figure 9), compared with the Model group, the cell morphology in the finasteride group was improved, the vesicular tubular structure was clear, and the thickness of prostatic epithelial cells was significantly reduced ( P < 0.05), and the proportion of the area of glandular epithelial cells in the visual field was also significantly reduced ( P < 0.01). A small number of internal folds protruded papillarily into the lumen, and the degree of protrusion was lower than that in the Model group; compared with the Model group, after TCS treatment, the cell morphology was significantly improved, the vesicular tubular structure was clearly visible, close to that of the control group, the internal folds in the acini were reduced, and the thickness of glandular epithelium was also significantly reduced ( P < 0.05). The proportion of the area of glandular epithelial cells in the visual field of the TCS-M and TCS-H groups was also significantly reduced, showing a highly significant difference ( P < 0.01). A small number of internal folds protruded papillarily into the lumen, and the degree of protrusion was significantly lower than that in the Model group; the cell morphology in the UA treatment group was also improved, the vesicular tubular structure was clearer than that in the Model group, and the thickness of epithelial cells was significantly reduced ( P < 0.05), and the proportion of the area of glandular epithelial cells was also significantly reduced compared with the Model group ( P < 0.01). Fewer internal folds protruded papillarily into the lumen, and the degree of protrusion was lower than that in the Model group; the above results suggest that TCS can significantly reduce the pathological state of prostatic hyperplasia and has a good therapeutic effect on lower urinary tract symptoms.

[0057] Determination of 5AR content in serum As Figure 10 known, compared with the Control group, the activity of 5AR in the Model group was significantly increased ( P < 0.01); compared with the Model group, the activities of 5AR in the finasteride group, TCS group and UA group were all significantly reduced ( P < 0.01), and the TCS group showed a dose-dependent effect. The effect of high-dose TCS was close to that of the positive drug finasteride. The above results indicate that TCS and UA can improve prostatic hyperplasia by inhibiting the activity of 5AR.

[0058] Effect of TCS on serum T and DHT contents in BPH rats Effect on serum T content Compared with the Control group, the content of testosterone (T) in the serum of rats in the Model group was significantly increased ( P < 0.01); compared with the Model group, the contents of T in the serum of rats in the finasteride group, TCS group and UA group were all significantly reduced ( P<0.01), and the medium and high doses of TCS had a better effect on T than the positive drug finasteride. As the dose of TCS increased, the content of T in the serum of BPH rats became lower, indicating that TCS had a dose-dependent effect on reducing the content of T. The experimental results showed that TCS and UA could improve the pathological state of prostatic hyperplasia by reducing the testosterone level in the body during benign prostatic hyperplasia ( Figure 11 ).

[0059] Effect on serum DHT content It can be seen from Figure 12 that compared with the Control group, the content of dihydrotestosterone (DHT) in the rats of the Model group increased significantly ( P <0.01); compared with the Model group, the DHT content in the rats of the finasteride group, TCS group and UA group decreased significantly ( P <0.01). As the dose of TCS increased, the DHT content also showed a gradually decreasing trend, indicating that TCS had a concentration-dependent effect on reducing the DHT content; and the effect of high-concentration TCS on reducing the DHT content was better than that of the positive drug finasteride. The change trend of DHT was consistent with that of T concentration, suggesting that TCS and UA reduced the content of DHT in the body while reducing the content of T in the body. TCS and UA could improve the pathological state of prostatic hyperplasia by simultaneously reducing the levels of DHT and T.

[0060] Effect of TCS on serum blood glucose level in BPH rats By measuring the blood glucose concentration in the serum of BPH rats, it was found that compared with the Control group, the blood glucose content in the BPH rats of the Model group induced by testosterone propionate increased significantly ( P <0.05); compared with the Model group, the finasteride group, TCS group and UA group could all reduce the blood glucose concentration in BPH rats. Among them, the blood glucose-lowering effect of UA was consistent with that of the positive drug at the same concentration, and it was statistically significant compared with the Model group ( P <0.05). It showed that ursolic acid, as the main component of TCS, could also treat BPH by reducing blood glucose ( Figure 13 ).

[0061] Effect of TCS on serum blood lipid level in BPH rats It can be seen from Figure 14 that compared with the Control group, the total cholesterol content in the Model group increased, indicating that the total cholesterol in the serum of rats showed an upward trend after modeling. The positive drug finasteride group was also slightly higher than the Control group, suggesting that finasteride might promote the synthesis of total cholesterol. Compared with the Model group, the total cholesterol levels in the TCS group and UA group decreased significantly ( P<0.01), indicating that UA and TCS have the effect of reducing total cholesterol at a certain dose.

[0062] Compared with the Control group, the triglyceride level in the Model group increased slightly, and the triglyceride content in the finasteride group was higher, suggesting that finasteride may promote the synthesis of triglyceride. Compared with the Model group, both the TCS group and the UA group had a reducing effect on triglyceride, and UA and high-dose TCS had a significant reducing effect on triglyceride ( P <0.01).

[0063] Compared with the Control group, the low-density lipoprotein (LDL) levels in the Model group and the finasteride group were both higher, and the high-density lipoprotein (HDL) level in the Model group decreased significantly. Compared with the Model group, the LDL levels in the TCS group and the UA group were lower, and the HDL levels increased, suggesting that TCS and UA have the effect of regulating LDL and HDL levels, but the effect of drug intervention on the recovery of HDL is relatively limited.

Claims

1. A 5α-reductase inhibitor, characterized in that, The inhibitor is total triterpenoids of Chaenomeles speciosa (Sweet) Nakai TCS.

2. The 5α-reductase inhibitor according to claim 1, characterized in that, The preparation method of the total triterpenoids of Chaenomeles speciosa (Sweet) Nakai TCS comprises the following steps: after the Fructus Chaenomelis is crushed, it is extracted with 60-95% ethanol as a solvent, concentrated, extracted with petroleum ether and ethyl acetate, the ethyl acetate extraction part is adsorbed by D101 macroporous adsorption resin, then eluted and separated with ethanol of different concentrations, and the 90-95% ethanol eluate is concentrated and dried after recovering ethanol to obtain the total triterpenoids of Chaenomeles speciosa (Sweet) Nakai.

3. Use of the 5α-reductase inhibitor according to claim 1 or 2 in the preparation of a medicament for treating prostatic hyperplasia.

4. Use of a total triterpenoids of Chaenomeles speciosa (Sweet) Nakai TCS in the preparation of a medicament for treating prostatic hyperplasia.

5. The application according to claim 4, characterized in that The use of the total triterpenoids of Chaenomeles speciosa (Sweet) Nakai TCS in the preparation of a medicament for treating prostatic hyperplasia is realized by reducing the total volume and wet weight of the prostate and seminal vesicles.

6. The application according to claim 4, characterized in that The use of the total triterpenoids of Chaenomeles speciosa (Sweet) Nakai TCS in the preparation of a medicament for treating prostatic hyperplasia is realized by improving cell morphology, making the vesicle tubular structure clearly visible, reducing the folds in the acinus, and reducing the thickness of the glandular epithelium.

7. The application according to claim 4, characterized in that, The use of the total triterpenoids of Chaenomeles speciosa (Sweet) Nakai TCS in the preparation of a medicament for treating prostatic hyperplasia is realized by reducing the increase in serum T level of BPH caused by exogenous testosterone propionate TP.

8. The application according to claim 4, characterized in that, The use of the total triterpenoids of Chaenomeles speciosa (Sweet) Nakai TCS in the preparation of a medicament for treating prostatic hyperplasia is realized by reducing the serum DHT level of BPH.

9. The application according to claim 4, characterized in that The use of the total triterpenoids of Chaenomeles speciosa (Sweet) Nakai TCS in the preparation of a medicament for treating prostatic hyperplasia is realized by reducing blood sugar and blood lipid.

10. The application according to any one of claims 4-9, characterized in that, The dosage of total triterpenoids TCS from papaya is 0.5 - 2.0 g·kg -1 .

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