阅读说明：本技术 一种联合用药物及用途 (Combined medicine and application ) 是由 苗琳 高秀梅 刘二伟 王亚静 常艳旭 马亚宣 吴垚锌 于 2021-06-28 设计创作，主要内容包括：本申请提供了一种联合用药物,其包括锁阳提取物和黄柏提取物,所述锁阳提取物单次用药与所述黄柏提取物单次用药的重量比为1:(1.0-1.5)。本申请还提供了所述联合用药物在制备预防和/或治疗前列腺增生并下尿路症状的药物中的用途。本申请提供的联合用药物,能够抑制前列腺体积的增大,降低前列腺指数和膀胱指数,改善前列腺和膀胱的组织病变,降低雌二醇/睾酮的水平,降低最大排尿压,升高逼尿肌肌条张力,从而能够用于预防和/或治疗前列腺增生并下尿路症状,进而用于制备预防和/或治疗前列腺增生并下尿路症状的药物。本申请的联合用药物具有协同增效的作用,适合长期治疗,临床应用前景良好。(The application provides a combined medicine, which comprises a cynomorium songaricum extract and a phellodendron extract, wherein the weight ratio of single medication of the cynomorium songaricum extract to single medication of the phellodendron extract is 1 (1.0-1.5). The application also provides the application of the combined medicine in preparing the medicine for preventing and/or treating prostatic hyperplasia and lower urinary tract symptoms. The combined medicine provided by the application can inhibit the enlargement of the prostate volume, reduce the prostate index and the bladder index, improve the tissue pathological changes of the prostate and the bladder, reduce the level of estradiol/testosterone, reduce the maximum urination pressure and increase the tension of detrusor muscle strips, so that the combined medicine can be used for preventing and/or treating the hyperplasia of the prostate and the lower urinary tract symptoms, and further can be used for preparing the medicine for preventing and/or treating the hyperplasia of the prostate and the lower urinary tract symptoms. The drug combination has a synergistic effect, is suitable for long-term treatment, and has a good clinical application prospect.)

1. A combination drug comprises a cynomorium songaricum extract and a phellodendron extract, wherein the weight ratio of single application of the cynomorium songaricum extract to single application of the phellodendron extract is 1 (1.0-1.5).

2. The combination as claimed in claim 1, wherein the cynomorium songaricum extract comprises at least one of an aqueous extract and an alcoholic extract of cynomorium songaricum.

3. The combination as claimed in claim 1, wherein the cortex Phellodendri extract comprises at least one of water extract and alcohol extract of cortex Phellodendri.

4. The combination according to claim 1, further comprising a pharmaceutically acceptable carrier or excipient.

5. The combination according to claim 4, wherein the pharmaceutically acceptable carrier or excipient is selected from at least one of a solvent, a diluent, a disintegrant, a precipitation inhibitor, a surfactant, a glidant, a binder, a lubricant, a dispersant, a suspending agent, an isotonicity agent, a thickener, an emulsifier, a preservative, a stabilizer, a hydrating agent, an emulsification accelerator, a buffer, an absorbent, a colorant, a flavoring agent, a sweetener, an ion exchanger, a mold release agent, a coating agent, a flavoring agent, or an antioxidant.

6. Use of a combination according to any one of claims 1 to 5 in the manufacture of a medicament for the prevention and/or treatment of prostatic hyperplasia and lower urinary tract symptoms.

7. The use according to claim 6, wherein the extract of Cynomorium songaricum and the extract of Phellodendri cortex are used simultaneously or sequentially in any order.

8. The use as claimed in claim 6, wherein the combination is for use in the order of administering the extract of Cynomorium songaricum first and then the extract of Phellodendri cortex.

9. The use of claim 8, wherein the cynomorium songaricum extract and the phellodendron amurense extract are used for 1-3h apart.

Technical Field

The application relates to the technical field of medicines, in particular to a combined medicine and application.

Background

Benign Prostatic Hyperplasia (BPH) is a highly pathogenic species of the urinary system of middle-aged and elderly men and is closely related to age. Epidemiological studies have shown that 50% of men over 60 years of age are able to observe BPH symptoms at the tissue level; the prevalence of BPH in men over 80 years of age approaches 90%. With the increasing of the volume of the prostate, the prostate can press the bladder neck to narrow the urethra, and finally Lower Urinary Tract Symptoms (LUTs) such as frequent micturition, urgent micturition, incomplete micturition and bladder outlet obstruction of patients with different degrees can occur, serious patients can have bilateral upper Urinary Tract hydrops and renal insufficiency, and serious complications such as hematuria, acute Urinary retention, bladder calculus, renal failure and prostate cancer can be easily caused if the treatment is not timely performed, and the life quality of men is seriously affected.

BPH patients are generally older, and considering factors such as physical quality of ill people and postoperative complications, drug therapy is the main treatment means of BPH. At present, the clinically common medicines for treating BPH comprise a 5 alpha-reductase inhibitor, an alpha-receptor blocker and the like, and the two medicines are usually combined clinically to treat prostatic hyperplasia and concurrent lower urinary tract symptoms, but the two medicines are found to relieve the BPH symptoms and simultaneously cause various adverse reactions such as sexual dysfunction, dizziness, hypotension and the like. Therefore, it becomes important to select safe and effective natural drugs with few side effects to inhibit prostatic hyperplasia and lower urinary tract symptoms.

Traditional Chinese medicine Cynomorium songaricum (Cynomorium songarium Rupr.) is a dry fleshy stem of Cynomorium songaricum of Cynomorium family, is mostly distributed in places such as internal Mongolia, Gansu and Qinghai, and is originally recorded in Ben Cao Yan Yi Bu Yi (supplement to the Yi) and has warm nature and sweet taste, and enters liver, kidney and large intestine meridians. Has effects of invigorating kidney yang, replenishing essence and blood, and loosening bowel to relieve constipation. Traditional Chinese medicine Phellodendron amurense (Phellodendron chinense Cortex) is a dry bark of Phellodendron chinense (Phellodendron chinense Schneid.) belonging to the rutaceae family, is known as Phellodendron amurense, and is originally recorded in Shennong Ben Cao Jing (Shennong herbal), and is cold in nature, bitter in taste, and belongs to kidney and bladder meridians. Has effects in clearing away heat, eliminating dampness, purging pathogenic fire, removing dampness, removing putrefaction, relieving swelling, removing toxic materials, and treating sore.

Disclosure of Invention

The present inventors have found, through intensive studies, that cynomorium songaricum and phellodendron amurense, in combination, are effective in inhibiting the enlargement of the prostate volume, reducing the prostate index and the bladder index, improving the histological changes of the prostate and bladder, and reducing the level of estradiol/testosterone, and thus can be used for preventing and/or treating prostatic hyperplasia and lower urinary tract symptoms, and have completed the present application based on this.

The first aspect of the application provides a combination drug, which comprises a cynomorium songaricum extract and a phellodendron extract, wherein the weight ratio of single medication of the cynomorium songaricum extract to single medication of the phellodendron extract is 1 (1.0-1.5).

A second aspect of the present application provides the use of a combination according to the first aspect of the present application for the manufacture of a medicament for the prevention and/or treatment of prostatic hyperplasia and lower urinary tract symptoms.

The combined medicine provided by the application can inhibit the enlargement of the prostate volume, reduce the prostate index and the bladder index, improve the tissue pathological changes of the prostate and the bladder, reduce the level of estradiol/testosterone, reduce the maximum urination pressure and increase the tension of detrusor muscle strips, so that the combined medicine can be used for preventing and/or treating the hyperplasia of the prostate and the lower urinary tract symptoms, and further can be used for preparing the medicine for preventing and/or treating the hyperplasia of the prostate and the lower urinary tract symptoms. The drug combination has a synergistic effect, is suitable for long-term treatment, and has a good clinical application prospect.

Drawings

In order to more clearly illustrate the embodiments of the present application or the technical solutions in the prior art, the drawings needed to be used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present application, and it is also obvious for a person skilled in the art to obtain other embodiments according to the drawings.

FIG. 1 shows the results of ultrasonic examination of prostate in each group of rats after administration.

FIG. 2 is a morphogram of prostate in each group of rats after administration.

Figure 3 is the prostate index results for each group of rats after dosing.

Figure 4 is the results of the bladder index of each group of rats after administration.

FIG. 5 shows the results of HE staining of prostate tissue in rats of each group after administration; wherein, the A picture is an HE staining picture of prostate tissues of rats in each group after administration, the upper picture of each group is a 10-fold under-lens result which is marked as 10X, and the lower picture is a 20-fold under-lens result which is marked as 20X; panel B shows the quantitative results of the thickness of columnar epithelium in Panel A.

Fig. 6 shows HE staining results of bladder tissue of rats in each group after administration, the upper panel of each group shows 4-fold under-the-mirror results and is labeled 4 ×, and the lower panel shows 10-fold under-the-mirror results and is labeled 10 ×.

FIG. 7 is the Masson staining of the bladder tissue of rats in each group after administration, wherein Panel A is the Masson staining of the bladder tissue of rats in each group after administration, and each panel is the result under a 10-fold microscope and is marked as 10 ×; and B is the result of the proportion of bladder fibrosis obtained from A.

Fig. 8 shows the results of the contents of estradiol, testosterone, estradiol/testosterone, and dihydrotestosterone in the serum of rats in each group after administration.

Figure 9 shows the maximum urinary pressure results for each group of rats after administration.

Figure 10 is the bladder detrusor muscle strip tension results for each group of rats after dosing.

Detailed Description

The technical solutions in the embodiments of the present application will be described clearly and completely with reference to the accompanying drawings, and it is obvious that the described embodiments are only a part of the embodiments of the present application, and not all of the embodiments. All other embodiments that can be derived by one of ordinary skill in the art from the description herein are intended to be within the scope of the present disclosure.

The first aspect of the application provides a combination drug, which comprises a cynomorium songaricum extract and a phellodendron extract, wherein the weight ratio of single medication of the cynomorium songaricum extract to single medication of the phellodendron extract is 1 (1.0-1.5).

In some embodiments of the first aspect of the present application, the cynomorium songaricum extract comprises at least one of an aqueous extract, an alcoholic extract of cynomorium songaricum. The extraction method of the cynomorium songaricum extract is reported in the prior art, and the cynomorium songaricum extract is not limited in the application as long as the purpose of the application can be achieved, and for example, the cynomorium songaricum extract can be obtained by extracting by at least one method of heating reflux, ultrasound, percolation, water decoction and the like. In the present application, the "alcohol extract" may be any one of a methanol extract, an ethanol extract, an aqueous methanol extract or an aqueous ethanol extract, and the present application is not limited thereto as long as the object of the present application can be achieved. In one embodiment, the cynomorium songaricum may be subjected to a heating reflux extraction with a 70-80% ethanol aqueous solution, to obtain a cynomorium songaricum extract, for example.

In some embodiments of the first aspect of the present application, the phellodendron extract comprises at least one of an aqueous extract, an alcoholic extract of phellodendron. The extraction method of the "phellodendron extract" described in the present application is reported in the prior art, and the present application does not limit this, as long as the object of the present application can be achieved, and for example, the "phellodendron extract" of the present application can be obtained by at least one method of heating reflux, ultrasound, percolation, water decoction, and the like. In one embodiment, phellodendron amurense may be subjected to ultrasonic extraction with methanol, to obtain an extract of phellodendron amurense.

In certain embodiments of the first aspect of the present application, the combination further comprises a pharmaceutically acceptable carrier or excipient.

Herein, "pharmaceutically acceptable" means having no substantial toxic effect when used in the usual dosage amounts, and thus being approved by the government or equivalent international organization or approved for use in animals, more particularly in humans, or registered in the pharmacopoeia.

The "pharmaceutically acceptable carrier or excipient" useful in the combination of the present application may be any conventional carrier in the art of pharmaceutical formulation and the selection of a particular carrier will depend on the mode of administration or the type and state of the disease used to treat a particular patient. The preparation of suitable combinations for a particular mode of administration is well within the knowledge of those skilled in the pharmaceutical art.

In some embodiments of the first aspect of the present application, the pharmaceutically acceptable carrier or excipient is selected from at least one of a solvent, diluent, disintegrant, precipitation inhibitor, surfactant, glidant, binder, lubricant, dispersant, suspending agent, isotonic agent, thickener, emulsifier, preservative, stabilizer, hydrating agent, emulsification accelerator, buffer, absorbent, colorant, flavoring agent, sweetener, ion exchanger, mold release agent, coating agent, flavoring agent, or antioxidant.

As used herein, the term "combination" has its ordinary meaning. In addition, the "combination drug" of the present application may also be present or provided in the form of a health product, a functional food, a food additive, or the like. The combination of the present application may be prepared by conventional techniques in the pharmaceutical field, in particular in the formulation field, by obtaining the active ingredients of the raw materials of the combination of the present application by extraction, separation and purification means commonly used in pharmaceutical manufacturing, optionally mixing with one or more pharmaceutically acceptable carriers or excipients, and then forming the desired dosage form. The combination according to the present application is a pharmaceutical preparation which can be suitably used for oral administration, a pharmaceutical preparation (e.g., solution) suitable for parenteral injection (e.g., intravenous injection, subcutaneous injection), a pharmaceutical preparation (e.g., ointment, patch or cream) suitable for surface administration, or a pharmaceutical preparation (e.g., suppository) suitable for rectal administration, and the like. Dosage forms for oral administration may include, for example, tablets, pills, hard or soft capsules, solutions, suspensions, emulsions, syrups, powders, fine granules, pellets, elixirs and the like, without limitation. In addition to the active ingredient, these preparations may contain diluents (e.g., lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, and glycine), lubricants (e.g., silica, talc, stearic acid or its magnesium salt, calcium salt, and polyethylene glycol). Tablets may also contain binders such as magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, and polyvinylpyrrolidone. If necessary, it may further contain pharmaceutically acceptable additives such as disintegrating agents (e.g., starch, agar, alginic acid or sodium salt thereof), absorbents, coloring agents, flavoring agents, sweetening agents, and the like. Tablets may be prepared according to conventional mixing, granulating or coating methods.

A second aspect of the present application provides the use of a combination according to the first aspect of the present application for the manufacture of a medicament for the prevention and/or treatment of prostatic hyperplasia and lower urinary tract symptoms.

The inventor finds in research that the combined medicament disclosed by the application can inhibit the enlargement of the volume of the prostate, reduce the prostate index and the bladder index, improve the tissue pathological changes of the prostate and the bladder, reduce the level of estradiol/testosterone, reduce the maximum micturition pressure and increase the tension of detrusor muscle strips, so that the combined medicament can be used for preventing and/or treating the hyperplasia of the prostate and the lower urinary tract symptoms, and further can be used for preparing the medicament for preventing and/or treating the hyperplasia of the prostate and the lower urinary tract symptoms.

The term "treatment" has its ordinary meaning in the present application and refers herein in particular to the treatment of a mammalian subject (preferably a human) already suffering from prostatic hyperplasia and lower urinary tract symptoms with a combination according to the present application in order to produce a therapeutic, curative, palliative etc. effect on said disease. Similarly, the term "prevention" as used herein has its ordinary meaning and herein refers in particular to the treatment of a mammalian subject who may or is at risk of suffering from prostatic hyperplasia and lower urinary tract symptoms with the combination of the present application in order to produce a preventing, arresting, abrupting, etc. effect on the disease.

In some embodiments of the second aspect of the present application, the extract of cynomorium songaricum and the extract of phellodendron amurense are used simultaneously or in any sequential order. In the present application, the term "combination" means that the cynomorium songaricum extract and the phellodendron amurense extract are used simultaneously or sequentially in any order to increase the efficacy of the drug and reduce the possibility of drug resistance in order to prevent and/or treat prostatic hyperplasia and lower urinary tract symptoms.

In some embodiments of the second aspect of the present application, the combination is administered in the order of the cynomorium songaricum extract first and the phellodendron amurense extract later. Preferably, the cynomorium songaricum extract and the phellodendron amurense extract are used for 1-3h at an interval.

The experimental materials and methods used in the following examples are, unless otherwise specified, conventional materials and methods.

Example 1 Effect of combination drug on prostate hyperplasia and amelioration of lower urinary tract symptoms

Preparing a cynomorium songaricum extract: extracting 3kg herba Cynomorii decoction pieces with 24L 60% ethanol under reflux for 2 times (each for 2 hr), evaporating the extractive solution in rotary evaporator, and lyophilizing in lyophilizer to obtain herba Cynomorii extract;

preparation of the phellodendron extract: extracting cortex Phellodendri decoction pieces 3kg with 30L 60% ethanol solution under heating and pressure for 2 times, each for 2 hr, mixing extractive solutions, filtering, distilling under reduced pressure to obtain extract, and vacuum drying to obtain cortex Phellodendri extract.

108 male Wistar rats without specific pathogen (SPF grade) were taken, weighed 230-250g, and castration surgery was started after adaptive feeding for one week. Randomly selecting 12 rats, performing a pseudo-operation on the rats, cutting the scrotum skin, the envelope tissue and the cremaster muscle of the anesthetized rats layer by layer in a sterile environment, drawing the testis, then sending the testis back into the envelope tissue, and suturing the skin layer by layer to serve as a pseudo-operation group; the rest 96 rats are castration operated, the bilateral testicles and epididymis are completely removed, a proper amount of sulfanilamide crystal powder is spread at the sutured position, and meanwhile, 0.3 mL/rat of gentamicin sulfate is injected intramuscularly to prevent wounds and postoperative infection.

The 96 rats after castration surgery are randomly divided into 8 groups, namely a model group, a cynomorium songaricum group, a phellodendron bark group, a combined drug 1 group, a combined drug 2 group, a combined drug 3 group, a combined drug 4 group and a tamsulosin group; the administration was started in groups at 21 days after the operation and continued for 28 days. The subcutaneous injections and the gavage administration are shown in Table 1. Among them, tamsulosin is a commonly used drug for treating prostatic hyperplasia and lower urinary tract symptoms at present, and is used as a positive control in the present application.

TABLE 1

The groups of rats were weighed after the last dose, and the prostate gland size was observed post-ultrasonically and the prostate volume calculated, the results are shown in figure 1. Fasting is carried out before material taking, 8 random rats are selected from each group for bladder perfusion detection of urination parameters, blood is taken from the posterior aorta, prostate and bladder tissues are taken simultaneously, and each tissue is cleaned, wherein the shape chart of the prostate of each group of rats is shown in figure 2. Accurately weighing the wet weight of each tissue by using an electronic balance to calculate the index of each organ of each group of experimental animals according to a formula: prostate wet weight/rat body weight × 100, Prostate Index (protate Index, PI, unit mg/100g body weight) was calculated, and the results are shown in fig. 3; according to the formula: the wet weight of the Bladder/rat body weight x 100, Bladder Index (Bladder Index, BI, in mg/100g body weight) was calculated and the results are shown in figure 4. Cutting prostate tissues at two sides, freezing the upper parts of the prostate tissues and the middle bladder tissues in liquid nitrogen, fixing the prostate dorsal lobe and the lower parts of the bladder in 4% paraformaldehyde, performing HE staining on the prostate tissues and the bladder tissues respectively by adopting a hematoxylin-eosin staining method (HE staining method) after paraffin embedding, and performing Masson staining on the bladder tissues by adopting a Masson staining method. The HE staining results of the prostate tissue of each group of rats are shown in a graph of fig. 5, in which the columnar epithelium of the prostate is shown by an arrow in a graph, and the thickness of the columnar epithelium of the prostate is quantified by Image J software, and the results are shown in B graph of fig. 5. The HE staining results of the bladder tissues of the rats in each group are shown in fig. 6, and the Masson staining results of the bladder tissues of the rats in each group are shown in a graph of fig. 7, and the area of collagen fibers and the total area of the bladder tissues in the a graph are measured by using Image J software according to the formula: the ratio of bladder fibrosis was obtained by dividing the area of collagen fibers by the total area of bladder tissue by 100%, and the results are shown in panel B of fig. 7. The other groups of rats were tested for detrusor contraction and relaxation by muscle strip experiments.

As can be seen from the prostate volume of each group of rats in FIG. 1, the prostate volume of the model group was significantly increased compared to that of the sham operation group; compared with the model group, the prostate volume of each administration group is obviously reduced. Wherein, under the condition that the dosage of the combined drug 1 group is half of the dosage of the cynomorium songaricum group and half of the dosage of the phellodendron amurense group, the prostate volume of the rat is equivalent to that of the cynomorium songaricum group and is smaller than that of the phellodendron amurense group; the combined drug 2 group had half the dose of the cynomorium songaricum group administered first and half the dose of the phellodendron amurense group administered 2 hours later, and the prostate volume of the rat was smaller than that of both the cynomorium songaricum group and the phellodendron amurense group, indicating that the combined drug of the present application has a synergistic effect. As can be seen from the morphology of the prostate gland in each group of rats in FIG. 2, the volume of the prostate gland in the rats in the model group was significantly increased compared to that in the sham-operated group; the prostate size was reduced to a different extent in each of the administration groups compared to the model group. The results show that the combined medicine can inhibit the enlargement of the prostate volume and has a synergistic effect; wherein the combined medicament of the cynomorium songaricum extract is applied firstly and the phellodendron extract is applied after 2 hours, so that the effect is better.

As can be seen from the prostate index results of fig. 3, the prostate index in the model group was significantly increased compared to the sham group (n ═ 8, ##### # P < 0.0001 compared to the sham group); compared with the model group, prostate indexes of the cynomorium songaricum group and the combined drug 2 group are remarkably reduced (n is 8, P is less than 0.05, P is less than 0.01, and compared with the model group), and the effect of the combined drug 2 group is stronger than that of the cynomorium songaricum group. As can be seen from the bladder index results of fig. 4, the bladder index of the model group was significantly increased compared to the sham group (n is 8, # # P < 0.001, compared to the sham group); bladder indexes of the cynomorium songaricum group and the combined drug 2 group are obviously reduced compared with those of the model group (n is 8, P is less than 0.01, compared with the model group), and the effect of the combined drug 2 group is equivalent to that of the cynomorium songaricum group. The results show that the combined medicine can reduce the prostate index and the bladder index and has a synergistic effect; wherein the combined medicament of the cynomorium songaricum extract is applied firstly and the phellodendron extract is applied after 2 hours, so that the effect is better.

According to the result of HE staining of the prostate tissue in FIG. 5, observing the change of the glandular cavities and cell forms of each group in the A picture of FIG. 5, the smooth muscle cell layer of the model group is obviously thickened, the arrangement of glandular epithelial cells is disordered, and the proliferation of connective tissues is more obvious; compared with the model group, the smooth muscle cell layer of each administration group is thinned, and the glandular epithelial cells are arranged neatly; as can be seen from the result of the thickness of the columnar epithelium of the prostate in the B-diagram of fig. 5, the thickness of the columnar epithelium of the prostate in the model group was significantly increased compared to the sham group (n 8, # # # # # # P < 0.0001 compared to the sham group); the thickness of the prostate columnar epithelium was significantly reduced in the combination 1 group compared to the model group (n-8, P < 0.05, compared to the model group). According to the HE staining results of bladder tissues in fig. 6 and Masson staining results in a panel a in fig. 7, it can be seen that the bladder wall was thickened and collagen fibers were increased in the model group; the bladder wall of each administration group becomes thin, and the collagen fiber is obviously reduced; as can be seen from the results of the bladder fibrosis ratio in the B-chart of fig. 7, the bladder fibrosis ratio in the model group was significantly increased compared to the sham operation group (n ═ 8, ##### # P < 0.0001 compared to the sham operation group); the proportion of cystic fibrosis was significantly reduced in both combination 1 and combination 3 compared to the model group (n 8, P < 0.01, P < 0.001 compared to the model group). The results show that the combination of the present application can improve the tissue pathology of prostate and bladder. From the results, the combined medicine has a synergistic effect, can obviously inhibit the enlargement of the prostate volume, obviously reduce the prostate index and the bladder index, and improve the tissue pathological changes of the prostate and the bladder.

Example 2 combination treatment of E in rat serum₂、T、E₂Influence of/T, DHT

The inventor finds that the imbalance of the androgen and estrogen level appears in the patients with prostatic hyperplasia and lower urinary tract symptoms, especially in clinic, E₂the/T is one of the current indexes for treating prostatic hyperplasia and lower urinary tract symptoms, and is generally regarded as E₂The lower the value of/T, the better the therapeutic effect of the drug. Estradiol (E) in rat serum₂) Testosterone (T), estradiol/testosterone (E)₂The content of the/T) and Dihydrotestosterone (DHT) is detected, so that the treatment effect of the combined medicament on the prostatic hyperplasia and lower urinary tract symptoms is verified.

In example 1, the abdominal aorta of each group of rats was bled after administration, centrifuged at 3500rpm/min for 10min to obtain the supernatant, and the serum levels of estradiol, testosterone, estradiol/testosterone, and dihydrotestosterone were measured in each group of rats, and the results are shown in fig. 8.

As can be seen from the results of fig. 8, the levels of estradiol and testosterone in the model group were significantly increased (n 8, # P < 0.01, in comparison with the sham group), the levels of testosterone were not significantly changed, the levels of estradiol and testosterone were significantly increased (n 8, # P < 0.01, in comparison with the sham group), and the levels of dihydrotestosterone were significantly increased (n 8, # P < 0.05, in comparison with the sham group). The combination 1, combination 2, combination 3 and combination 4 all had significantly reduced estradiol levels (n-8, P < 0.05, P < 0.001, P < 0.0001, compared to the model group), the testosterone levels in each group did not change significantly, the increase in estradiol/testosterone was significantly inhibited in each group (n-8, P < 0.05, P < 0.0001, compared to the model group), the combination 2, combination 3 and combination 4 all had significantly reduced dihydrotestosterone levels (n-8, P < 0.05, P < 0.001, P < 0.0001, compared to the model group); in the results of the estradiol/testosterone content levels, the combined drug 3 groups were significantly reduced compared with the cynomorium songaricum group (n is 8, Δ P < 0.01, compared with the cynomorium songaricum group), and the combined drug 3 groups were also significantly reduced compared with the phellodendron amurense group (n is 8, Δ P < 0.05, compared with the phellodendron amurense group), which indicates that the combined drug of the present application is E-shaped after administration₂Has low content of/T and good therapeutic effect on prostatic hyperplasiaThe synergistic effect of the cynomorium songaricum and the phellodendron bark is shown in the combined medicine.

Example 3 Effect of combination on maximum urinary pressure in rats

In example 1, the urinary parameters were measured by performing bladder perfusion on each group of rats, and the measurement result of the maximum urinary pressure (MVP) of each group of rats is shown in FIG. 9, wherein the bladder perfusion is performed by injecting the abdominal cavity of each group of rats for anesthesia, fixing the rats in the upward position, and pressing the lower abdomen of the rats to discharge residual urine in the bladder; opening the abdominal cavity to separate and cut off the ureter, and ligating the ureter close to the bladder end; the venous needle is inserted from the top of the bladder, the other side of the venous needle is connected with a hose connected with a micro-perfusion pump, and signals are collected and recorded by a four-channel pressure sensor PowerLab 8/30. Background levels of pressure in the rat bladder were set to zero prior to the experiment. The bladders were perfused at room temperature with 0.9% physiological saline through a micro-perfusion pump at a rate of 0.17ml/min, and the MVP was determined, with three consecutive recordings per rat. From the results in fig. 9, it can be seen that the maximal micturition pressure was significantly increased in the model group compared to the sham group (n 8, # # P < 0.001, compared to the sham group), while both the combination 1 and 2 groups were able to significantly lower the maximal micturition pressure in the rats (n 8, # P < 0.001, # P < 0.0001, compared to the model group). The results show that the combination of the present application can significantly reduce the maximum urinary pressure.

Example 4 Effect of combination drug on rat bladder detrusor muscle strip tension

The results of the muscle strip test in example 1, in which the abdominal cavity of each group of rats was anesthetized and 1 minute later, the muscle strip test was performed on the detrusor muscle of each group of rats, and the tension of the muscle strip of the detrusor muscle of each group of rats was measured, are shown in FIG. 10: opening the abdominal cavity, quickly taking out the bladder, placing the bladder in Krebs-Henseleit (K-H) liquid which is precooled on ice in advance and presaturated by introducing oxygen, removing connective tissues around the bladder as much as possible, cutting 4 longitudinal muscle strips (about 6mm multiplied by 2 mm/strip) along the traveling direction of muscle fibers, fixing two ends of each muscle strip on a flat dish with a rubber bottom by using acupuncture needles, tying one end of each muscle strip on an L-shaped stainless steel small hook by using an operation line, tying the other end of each muscle strip on a homemade operation line hook (which is convenient for subsequent adjustment and hanging on a tensioner) (one end of the operation line is a small hook made of a staple, and the other end of each operation line is a loop tied and connected with the tensioner); the prepared muscle strips are placed in a tissue bath, the end connected with an L-shaped stainless steel hook is arranged at the bottom, the other end is hung on a tensioner through a wire loop, and isometric contraction activities of the muscle strips are recorded by a PowerLab 8/35 biological signal processing system. As can be seen from the results of fig. 10, the muscle tone of the model group rats was significantly decreased compared to the sham group (n ═ 6, # # P < 0.01, compared to the sham group); and the combined drug 1 group, the combined drug 2 group and the combined drug 3 group can obviously increase the muscle tension of rats in each group (n is 6, P is less than 0.001, P is less than 0.0001 compared with the model group). The results show that the drug combination of the application can obviously increase the tension of detrusor muscle strips.

In conclusion, when half dose of cynomorium songaricum and half dose of phellodendron amurense are simultaneously administrated, the medicine has better improvement effect on the thickness of columnar epithelium of prostate and the estradiol level than that of cynomorium songaricum or phellodendron amurense which is singly administrated, and has better improvement effect on the fibrosis ratio of bladder and the detrusor muscle strip tension than that of cynomorium songaricum which is singly administrated; when half of the cynomorium songaricum is administered firstly and half of the phellodendron amurense is administered after 2 hours, the medicine has better improvement effect on the estradiol level and the dihydrotestosterone level than the single administration of the cynomorium songaricum or the phellodendron amurense, better improvement effect on the tension of detrusor muscle strips than the single administration of the cynomorium songaricum, and better improvement effect on the prostate index and the bladder index than the single administration of the phellodendron amurense; when half dose of phellodendron amurense is applied firstly, and half dose of cynomorium songaricum is applied for administration after 2 hours, the better improvement effect on the estradiol level, the estradiol/testosterone level and the dihydrotestosterone level than the single administration of cynomorium songaricum or phellodendron amurense is achieved, and the better improvement effect on the bladder fibrosis ratio and the detrusor muscle strip tension than the single administration of cynomorium songaricum is achieved; when half of the cynomorium songaricum is firstly applied and half of the phellodendron amurense is applied for administration after 8 hours, the effect of improving the estradiol level and the dihydrotestosterone level is better than that of singly applying the cynomorium songaricum or the phellodendron amurense. Therefore, compared with the single medicine of cynomorium songaricum or phellodendron amurense, the combined medicine can more effectively improve different pathological manifestations of prostatic hyperplasia and lower urinary tract symptoms, and the combined medicine has better treatment effect than the single medicine of cynomorium songaricum or phellodendron amurense when treating the prostatic hyperplasia and lower urinary tract symptoms, and is further used for preparing the medicine for preventing and/or treating the prostatic hyperplasia and lower urinary tract symptoms.

The above description is only for the preferred embodiment of the present application and is not intended to limit the scope of the present application. Any modification, equivalent replacement, improvement and the like made within the spirit and principle of the present application are included in the protection scope of the present application.