Traditional Chinese medicine composition for treating chronic prostatitis and preparation method thereof

Abstract

The invention belongs to the technical field of traditional Chinese medicine, and particularly discloses a traditional Chinese medicine composition for treating chronic prostatitis and a preparation process thereof.The traditional Chinese medicine composition is prepared from, by weight, 3-15 parts of cassia twigs, 20-40 parts of oysters, 20-40 parts of fossil fragments, 10-30 parts of yam rhizome, 10-30 parts of pyrrosia lingua and 20-40 parts of salvia miltiorrhiza. The traditional Chinese medicine composition has the effects of clearing heat and promoting diuresis, dissolving turbidity and treating stranguria, promoting blood circulation to remove blood stasis, and resolving depression for tranquilization, so that a good effect of treating chronic prostatitis caused by damp-heat stasis is achieved, and the traditional Chinese medicine composition is convenient to take, small in adverse reaction and wide in application prospect.

Description

Technical Field

[0001] The invention relates to a traditional Chinese medicine composition, in particular to a traditional Chinese medicine composition for treating prostate diseases, belonging to the technical field of traditional Chinese medicines. Background Art

[0002] Chronic prostatitis (CP) is a common inflammatory disease of the reproductive system in young and middle-aged men, including type II chronic bacterial prostatitis (CBP) and type III chronic prostatitis / chronic pelvic pain syndrome. According to statistics, about 50% of men will suffer from prostatitis at some stage in their lives. The incidence of chronic prostatitis reported in China is 6% to 32.9%. There is no term "prostate" in traditional Chinese medicine. Doctors of all dynasties have classified the prostate as "essence chamber, essence orifice, and essence channel". Prostatitis belongs to the category of "essence turbidity", "stranguria" and "leucorrhea" in traditional Chinese medicine. Traditional Chinese medicine believes that chronic prostatitis has fewer single pathogenesis, and most of them are caused by complex factors, such as damp heat, qi stagnation, blood stasis, and deficiency of vital energy. Damp heat and turbid evil are internally accumulated or deposited, invading the liver and kidney in the lower part of the body, accumulating in the essence chamber, and blocking the liver meridian over time, causing qi stagnation and blood stasis, and emotional distress.

[0003] Infection, immune factors, mental and psychological factors, pelvic floor dysfunction and other factors can all lead to chronic prostatitis, and the role of mental and psychological factors in the onset of the disease should be paid special attention to. Clinically, it is characterized by slow onset, stubborn disease, repeated attacks, and lingering and difficult to cure. The main symptoms of chronic prostatitis include pelvic pain symptoms, lower urinary tract symptoms (LUTS), mental and psychological symptoms, and sexual dysfunction symptoms. Due to the complex etiology, diverse symptoms, and different diagnosis and treatment of chronic prostatitis, the treatment is mainly to relieve clinical symptoms and improve the quality of life of patients. Commonly used Western medicines in clinical practice have their limitations. For example, α-receptor blockers are the first choice for relieving LUTS. They can improve patients' pain and urination symptoms and have a mild effect, but they cannot be used for a long time. Antibiotics are empirical treatment drugs. Long-term use can cause adverse reactions such as liver damage, intestinal discomfort, and crystalluria. Non-steroidal anti-inflammatory drugs (NSAIDs) can only be used for early short-term pain due to adverse reactions. Western medicine has no targeted treatment for anxiety and depression, and it is difficult to relieve patients' related mental symptoms. Several diagnosis and treatment guidelines / expert consensus issued in recent years have emphasized the role of traditional Chinese medicine in the diagnosis and treatment of chronic prostatitis. Summary of the invention

[0004] The present invention provides a Chinese medicine composition for treating chronic prostatitis and a preparation method thereof. The composition is modified from Guizhi Longgu Muli Decoction, and the raw materials are composed of Guizhi, oyster, dragon bone, Radix Dichroae, Pyrrosiae and Salvia miltiorrhiza. In the prescription, Radix Dichroae and Pyrrosiae enter the kidney and bladder meridians, clear away heat and dampness, clear and remove turbidity, and promote diuresis and relieve stranguria, and are the main medicines; Salvia miltiorrhiza promotes blood circulation and removes blood stasis, clears the heart and removes vexation, and dragon bone and oyster calm the nerves, soften and disperse nodules, and are the ministerial medicines, so as to treat the combined symptoms of blood stasis and emotional depression; Guizhi is used to warm the spleen and kidney to help qi transformation, and help the two main and ministerial medicines to remove dampness and remove turbidity, and promote blood circulation and remove blood stasis. All the medicines are used together to play the functions of clearing heat and dampness, removing turbidity and relieving stranguria, promoting blood circulation and removing blood stasis, and relieving depression and calming the nerves, and can be used to treat chronic prostatitis caused by damp-heat and blood stasis.

[0005] One of the purposes of the present invention is to provide a traditional Chinese medicine composition for treating prostate diseases, characterized in that the traditional Chinese medicine composition is mainly prepared from the following raw materials: cinnamon twig, oyster, dragon bone, radix polygoni multiflori, pyrrosia, and salvia miltiorrhiza.

[0006] In some embodiments, the Chinese medicine composition is prepared from the following raw materials: 3-15 parts by weight of cinnamon twig, 20-40 parts by weight of oyster, 20-40 parts by weight of dragon bone, 10-30 parts by weight of radix polygoni multiflori, 10-30 parts by weight of pyrrosia, and 10-40 parts by weight of salvia miltiorrhiza.

[0007] In a more preferred embodiment, the Chinese medicine composition is prepared from the following raw materials: 15 parts by weight of cinnamon twig, 40 parts by weight of oyster, 40 parts by weight of dragon bone, 30 parts by weight of radix polygoni multiflori, 30 parts by weight of pyrrosia, and 40 parts by weight of salvia miltiorrhiza.

[0008] In a more preferred embodiment, the Chinese medicine composition is prepared from the following raw materials: 6 parts by weight of cinnamon twig, 30 parts by weight of oyster, 30 parts by weight of dragon bone, 20 parts by weight of radix polygoni multiflori, 20 parts by weight of pyrrosia, and 30 parts by weight of salvia miltiorrhiza.

[0009] The second object of the present invention is to provide the use of the above-mentioned Chinese medicine composition in treating prostate diseases, wherein the prostate diseases are one or more of prostatitis, prostate hyperplasia and prostate cysts.

[0010] In some embodiments, the prostatitis is one or more of acute bacterial prostatitis, chronic bacterial prostatitis, chronic prostatitis / chronic pelvic pain syndrome, and asymptomatic prostatitis.

[0011] In some embodiments, the prostatitis is divided into damp-heat invasion type, damp-heat stasis type, qi stagnation and blood stasis type, yin deficiency and hyperactivity of fire type, kidney yang deficiency type or spleen deficiency and dampness type according to TCM syndrome differentiation.

[0012] The third aspect of the present invention is to provide a method for preparing a traditional Chinese medicine composition for treating prostate diseases.

[0013] In some embodiments, the method for preparing the Chinese medicine composition adopts the following method:

[0014] (1) calcining dragon bones and oysters, crushing and mixing to obtain a mixed powder, sieving, and subjecting the mixed powder to decoction extraction or reflux extraction to obtain an extract;

[0015] (2) extracting Danshen with 40-95% ethanol to obtain an ethanol extract;

[0016] (3) extracting Radix Dioscoreae and Pyrola with water to obtain a water extract;

[0017] (4) filtering the extract of step (3), concentrating under reduced pressure, adding ethanol and filtering to obtain a filtrate;

[0018] (5) Mix the extracts from steps (1), (2), and (4).

[0019] In a more preferred embodiment, the method for preparing the Chinese medicine composition adopts the following method:

[0020] (1) The dragon bone and oyster are crushed and mixed to obtain a mixed powder, and the mixed powder is sieved. The mixed powder is mixed with water at a ratio of 1:10, and the mixture is boiled at 95-100° C. for 1 hour. After filtering, the decoction is considered to have completed one extraction. The extraction is repeated 3-4 times and the decoction is combined;

[0021] (2) extracting Danshen with 50-80% ethanol to obtain an ethanol extract;

[0022] (3) extracting Radix Dioscoreae and Pyrola with water to obtain a water extract;

[0023] (4) filtering the water extract of step (3), concentrating under reduced pressure to a relative density of 1.05 to 1.25 at 50° C., adding ethanol to make the alcohol content reach more than 40%, and filtering to obtain a filtrate;

[0024] (5) Mix the extracts from steps (1), (2), and (4).

[0025] A fourth object of the present invention is to provide a Chinese medicine oral preparation.

[0026] In some embodiments, the oral preparation is one or more of pills, granules, tablets, pellets, micropills, and capsules.

[0027] Compared with the prior art, the present invention has achieved remarkable technical effects:

[0028] 1) The Chinese medicine composition of the present invention can significantly improve the prostate index of rats with chronic non-bacterial prostatitis, reduce the level of prostate inflammation, reshape the pathological morphology of prostate tissue, reduce the level of tissue fibrosis, relieve the anxiety behavior of rats, and lower the pain threshold.

[0029] 2) The present invention can reduce the prostate index of rats with chronic bacterial prostatitis, improve the swelling and inflammatory state of prostate tissue, significantly reduce the number of bacterial colonies in rat prostate tissue, alleviate the level of prostate inflammation, reshape the pathological morphology of prostate tissue, reduce the degree of chronic inflammation, reduce the tissue collagen volume fraction of rats, and reduce the number of white blood cells, neutrophils, and lymphocytes in rat blood routine tests.

[0030] In addition, there was no significant change in the blood routine indicators of rats with chronic prostatitis after treatment, and the safety was good. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 In: a. The effect of the Chinese medicine composition of the present invention on the pain threshold of rats with chronic nonbacterial prostatitis; b. The effect of the Chinese medicine composition of the present invention on the pain anxiety of rats with chronic nonbacterial prostatitis;

[0032] Figure 2 Effect of the Chinese medicine composition of the present invention on the prostate index of rats with chronic nonbacterial prostatitis;

[0033] Figure 3 Effects of the Chinese medicine composition of the present invention on TNF-α, IL-1β and IL-6 in rats with chronic nonbacterial prostatitis;

[0034] Figure 4 Effect of the Chinese medicine composition of the present invention on the pathological morphology of prostate tissue in rats with chronic non-bacterial prostatitis (H&E staining);

[0035] Figure 5 Effect of the Chinese medicine composition of the present invention on fibrosis in rats with chronic nonbacterial prostatitis (Masson staining);

[0036] Figure 6 Effect of the Chinese medicine composition of the present invention on inflammatory factor TNF-α in prostate tissue of rats with chronic bacterial prostatitis;

[0037] Figure 7 Effect of the Chinese medicine composition of the present invention on inflammatory factor IL-1β in prostate tissue of rats with chronic bacterial prostatitis;

[0038] Figure 8 Effect of the Chinese medicine composition of the present invention on prostate histopathology in rats with chronic bacterial prostatitis (H&E staining);

[0039] Fig. 9 Effect of the Chinese medicine composition of the present invention on collagen fiber staining of prostate tissue in rats with chronic bacterial prostatitis (Masson staining);

[0040] Fig.10The influence of the Chinese medicine composition of the present invention on the collagen fiber volume fraction of the prostate tissue of rats with chronic bacterial prostatitis. DETAILED DESCRIPTION

[0041] The present invention is further described below by way of examples. It should be correctly understood that the examples of the present invention are only some representative formulations, and their purpose is only to illustrate the present invention, rather than to limit the present invention. Therefore, simple improvements to the present invention under the premise of the method of the present invention all belong to the scope of protection claimed in the present invention.

[0042] Example 1: A Chinese medicine composition for treating chronic prostatitis

[0043] 6 parts by weight of cinnamon twig 30 parts by weight of oyster 30 parts by weight of dragon bone 20 parts by weight of pyrrosia 20 parts by weight of salvia miltiorrhiza 30 parts by weight

[0044] (1) The dragon bone and oyster are crushed and mixed to obtain a mixed powder, and the mixed powder is sieved. The mixed powder is mixed with water at a ratio of 1:10, and the mixture is boiled at 95-100° C. for 1 hour. After filtering, the decoction is considered to have completed one extraction. The extraction is repeated 3-4 times and the decoction is combined;

[0045] (2) extracting Danshen with 50-80% ethanol to obtain an ethanol extract;

[0046] (3) extracting Radix Dioscoreae and Pyrola with water to obtain a water extract;

[0047] (4) filtering the water extract of step (3), concentrating under reduced pressure to a relative density of 1.05 to 1.25 at 50° C., adding ethanol to make the alcohol content reach more than 40%, and filtering to obtain a filtrate;

[0048] (5) The extracts of steps (1), (2) and (4) are mixed to obtain a decoction.

[0049] Example 2: A Chinese medicine composition for treating chronic prostatitis

[0050] 3 parts by weight of cinnamon twig 20 parts by weight of oyster 20 parts by weight of dragon bone 10 parts by weight of pyrrosia 10 parts by weight of salvia miltiorrhiza 20 parts by weight

[0051] Preparation method: Same as Example 1.

[0052] Example 3: A Chinese medicine composition for treating chronic prostatitis

[0053] 15 parts by weight of cinnamon twig, 40 parts by weight of oyster, 2 parts by weight of dragon bone, 30 parts by weight of radix polygoni multiflori, 30 parts by weight of pyrrosia, and 40 parts by weight of salvia miltiorrhiza.

[0054] Preparation method: Same as Example 1.

[0055] Comparative Example 1: A Chinese medicine composition for treating chronic prostatitis

[0056] 20 parts by weight of cinnamon twig, 15 parts by weight of oyster, 15 parts by weight of dragon bone, 8 parts by weight of radix polygoni multiflori, 8 parts by weight of pyrrosia, and 8 parts by weight of salvia miltiorrhiza.

[0057] Crush the dragon bones and oysters, mix them to get the mixed powder, sieve it, and mix it with the other four medicines, use seven liters of water, boil it to get three liters, and filter it.

[0058] Comparative Example 2: A Chinese medicine composition for treating chronic prostatitis

[0059] 6 parts by weight of cinnamon twig, 30 parts by weight of oyster, 30 parts by weight of dragon bone, 20 parts by weight of sarsaparilla, 20 parts by weight of plantain seeds, 9 parts by weight of ginger, 6 parts by weight of licorice, and 5 jujubes.

[0060] Crush the dragon bones and oysters, mix them to get a mixed powder, sieve it, and mix it with the other six medicines, use seven liters of water, boil it to get three liters, and filter it.

[0061] Comparative Example 3: A Chinese medicine composition for treating chronic prostatitis

[0062] 3g cinnamon twig, 3g peony root, 3g ginger, 2g licorice, 12 dates, 3g dragon bone, and 3g oyster.

[0063] Take the above seven herbs, boil them in seven liters of water and take out three liters, then filter to get a decoction.

[0064] Pharmacodynamics test

[0065] Experimental Example 1 Study on the therapeutic effect of the Chinese medicine composition of the present invention on chronic non-bacterial prostatitis

[0066] 1. Establishment of rat model of chronic nonbacterial prostatitis

[0067] The chronic non-bacterial prostatitis model was induced by 3% carrageenan solution. After 7 days of adaptive feeding, all Sprague Dawley (SD) rats were anesthetized by intraperitoneal injection of 40 mg / kg sodium pentobarbital. The lower abdomen was shaved to expose the skin. Under anesthesia, the surgical incision was exposed for 1.5 cm, and the ventral lobe of the prostate was found. Each lobe was injected with 3% carrageenan solution twice, 50 μl each. The sham operation group was injected with normal saline, and wiped with cotton balls after injection to prevent leakage. After the injection, the prostate was put back and sutured with a suture needle. Penicillin (40U / L, 10,000 units) was injected intraperitoneally after surgery to prevent infection.

[0068] 2. Experimental Grouping and Drug Administration

[0069] Male SD rats (200±20 g) were randomly divided into 7 groups, with 10 rats in each group: sham operation group (Sham), model group (Model), low-dose group (Low) of Example 1, medium-dose group (Medium) of Example 1, high-dose group (High) of Example 1, Chinese medicine control group (QLX), and Western medicine control group (SLXB).

[0070] The dosage of the rats in the low-dose group of Example 1 was 12.24 g crude drug / kg body weight, the dosage of the rats in the medium-dose group of Example 1 was 36.72 g crude drug / kg body weight, and the dosage of the rats in the high-dose group of Example 1 was 110.16 g crude drug / kg body weight, and the dosage volume was 0.1 ml / 10 g. The Chinese medicine control group was administered 0.54 g / kg, and the Western medicine control group was administered 0.018 g / kg. After the 7th day of modeling, each administration group was continuously administered for 21 days, once a day, and the sham operation group and the model group were given normal saline every day, and the administration volume was 0.1 ml / 10 g.

[0071] 3. Determination of detection indicators

[0072] 3.1 Pain threshold and open field test

[0073] The pain threshold can directly reflect the pain-related behavioral changes in rats. Using a thermal pain meter, plantar pain tests were performed on the 7th, 14th, 21st, and 28th days after rat modeling. The rats were placed on a hot plate preheated to (55±0.5)°C. After the temperature stabilized, the rats were placed on the hot plate and timing began. The timing stopped when the rats first licked their hind paws, with a cutoff time of 60 seconds. Repeat 3 times and take the average value, with an interval of 20 minutes between each measurement.

[0074] The open field test can determine the anxiety behavior of rats. The operator holds the proximal end of the rat's tail and gently places it into the central grid of the open field test box, and records the behavior within 5 minutes. The movement trajectory of the mouse is recorded by a camera and analyzed using tracking software. The main observation indicator is the tactile index (tactile index = distance of the rat in the surrounding area / total distance). After the experiment, the feces of each animal will be thoroughly cleaned, and then the bottom of the cage will be sprayed with 75% ethanol and wiped dry with clean gauze to avoid any potential impact on subsequent experiments.

[0075] 3.2 Prostate Index

[0076] The prostate index is an important factor in evaluating the development of prostatitis and reflects the degree of prostate inflammation. The prostate index increases with the progression of inflammation. Rats were anesthetized with 10% chloral hydrate, the abdominal cavity was opened under sterile conditions, the prostate tissue was taken out, cleaned, and the prostate tissue was weighed. The prostate visceral coefficient was calculated according to the following formula.

[0077] Prostate index = total wet weight of prostate tissue / rat body weight.

[0078] 3.3ELISA method to detect the expression of inflammatory factors TNF-α, IL-1β, and IL-6 in prostate tissue

[0079] TNF-α, IL-1β and IL-6 are considered to be important proinflammatory factors, playing an important role in the occurrence and development of prostatitis. TNF-α and IL-6 can be used as indicators of the strength of local immune response in the prostate, diagnosis of chronic prostatitis and disease detection. Reducing the level of proinflammatory cytokine TNF-α is of great significance in preventing organ damage. Therefore, the ELISA method was selected to detect the expression of tissue inflammatory factors TNF-α, IL-6 and IL-1β.

[0080] 3.4 Pathological tissue morphology analysis

[0081] H&E staining can directly reflect the changes in tissue pathological morphology, and is also the most intuitive indicator for diagnosing prostatitis. The prostate tissue of the same area of ​​each rat was fixed in 4% tissue fixative, dehydrated, routinely paraffin-embedded and sectioned, with a thickness of 5μm, and H&E staining for histological scoring. Five random fields were selected for each sample under an optical microscope, and the severity of prostatitis was scored according to the chronic inflammatory cell infiltration and glandular epithelial thickness in each area.

[0082] The scoring criteria are as follows: 1 point - inflammatory cells are scattered, with 1 to 10 inflammatory cells / high-power microscope (HP×400); 2 points - inflammatory cells are aggregated, with no glandular epithelial tissue destruction or lymphoid nodules / follicles formation, with 11 to 20 inflammatory cells / HP; 3 points - inflammatory cells are aggregated, with partial glandular epithelial tissue destruction or lymphoid nodules / follicles formation, with inflammatory cells >20 / HP; 4 points - inflammatory cells are aggregated, with a large amount of glandular epithelial tissue destruction or lymphoid nodules / follicles formation, and the number of inflammatory cells fills the entire field of view / HP.

[0083] 3.5 Collagen fiber volume analysis

[0084] Masson staining can detect collagen deposition in rat prostate tissue, and tissue sections are stained according to the instructions. Before staining, the paraffin sections of mouse prostate tissue are dewaxed, and the dewaxing steps are the same as those before H&E staining.

[0085] The specific steps of Masson staining are as follows: (1) Place the slices in distilled water to moisten for 30 to 60 seconds, then place them in nuclear staining solution for 1 to 2 minutes, slowly pour out the nuclear staining solution, rinse the slices with a pre-prepared weak acid rinse solution for about 30 seconds, and wash away the residual nuclear staining solution; (2) Place the slices in pulp staining solution for staining, let it stand for about 1 minute, slowly pour out the pulp staining solution, rinse with a weak acid rinse solution for about 30 seconds, and wash away the residual pulp staining solution; (3) Place the slices in color separation solution, let it stand for 6 to 8 minutes, and slowly pour out the color separation solution; (4) Place the slices in blue counterstain solution, let it stand for about 5 minutes, and slowly pour out the blue counterstain solution; (5) Place the slices in 95% ethanol solution for rapid dehydration for 5 to 10 seconds, and then repeatedly dehydrate and rinse with anhydrous ethanol; (6) After blowing the slices dry, seal them with neutral gum and observe them under a microscope.

[0086] 3.6 Data processing

[0087] The data were processed using SPSS 22.0 software. The data were compared among the groups using one-way analysis of variance and multiple comparisons, and the graphs were drawn using Sigma Plot 14. P < 0.05 indicated that the difference was statistically significant.

[0088] 4. Experimental Results

[0089] 4.1 Effects of Chinese herbal medicine combination on pain threshold and anxiety

[0090] The pain threshold can directly reflect the pain changes of rats and indirectly reflect the changes in the inflammation level of rats. The pain threshold results are shown in Table 1 and Figure 1 -a.

[0091] Compared with the control group, the pain threshold of the model group decreased significantly. At the same time, the pain threshold of the rats in the model group was maintained at a low level on the 14th, 21st and 28th days, indicating that the prostatitis rat model can be maintained for at least 28 days (p<0.05). After administration on the 7th day, the pain threshold of the rats in the administration group generally increased. After the 21st day, the pain threshold of the rats in the group administered with the Chinese medicine composition of the present invention showed a dose-dependent increase. Among them, the pain threshold of the high-dose rats increased over time, indicating that the high-dose had a better therapeutic effect and had a more obvious analgesic effect (P<0.05). The Western medicine control group relieved the pain of rats in a time-dependent manner, and also showed a good effect on the 28th day (P<0.05).

[0092] The open field test can reflect the mental state and anxiety of rats. Figure 1-b, compared with the control group, the tactile index of the rats in the model group increased significantly, indicating that the number of times and distances of rats entering the central area decreased, and most of them were active in the surrounding area, indicating that the exploration desire of the model rats decreased and more obvious anxiety behavior appeared (P<0.05). Compared with the model group, the tactile index of the rats in the high-dose group of the Chinese medicine composition of the present invention was significantly reduced, indicating that the anxiety behavior of the rats was relieved after administration, showing a higher desire for exploration (p<0.01). The anxiety behavior of the rats in the western medicine control group was also well relieved (P<0.05).

[0093] Table 1 Changes in pain threshold of rats in each group at different time points

[0094]

[0095] Compared with the Sham group, the Model group # P<0.05, ## P<0.01, ### P < 0.001;

[0096] Compared with the Model group, *P<0.05, **P<0.01, ***P<0.01

[0097] 4.2 Effects of Chinese herbal medicine combination on prostate index

[0098] like Figure 2 As shown, compared with the control group, the prostate index of rats in the model group was significantly increased (P<0.01), indicating that the prostate of rats in this disease state had obvious inflammatory symptoms.

[0099] Compared with the model group, the prostate index of rats in the low-dose and medium-dose groups of the Chinese medicine composition of the present invention has a downward trend, but there is no statistical significance (P>0.05); the prostate index of rats in the high-dose group of the Chinese medicine composition of the present invention has a significant decrease (P<0.01), indicating that the high-dose group of the Chinese medicine composition of the present invention can significantly improve the prostate index of rats and gradually reduce prostate inflammation in rats in a dose-dependent manner, and the high-dose Chinese medicine has the most significant effect. Both the Chinese medicine control group and the Western medicine control group have a more obvious downward trend, among which the Western medicine control group has a better effect (P<0.05).

[0100] 4.3 Effects of Chinese herbal medicine combination on inflammatory factors in prostate tissue

[0101] like Figure 3 As shown in the figure, compared with the control group, the inflammatory factors TNF-α, IL-1β, and IL-6 in the model group were significantly increased (P<0.05).

[0102] Compared with the model group, the TNF-α levels of rats in the low-dose and medium-dose groups of the Chinese medicine composition of the present invention showed a significant downward trend (p<0.01), and the high-dose group decreased more, indicating that the high dose has a better anti-inflammatory effect (P<0.001). The TNF-α levels were also reduced in the Chinese medicine control group (P<0.05) and the Western medicine control group (P<0.01).

[0103] Compared with the model group, the low-dose, medium-dose and Chinese medicine control groups of the Chinese medicine composition of the present invention had no significant difference in IL-1β levels. The Chinese medicine composition of the present invention reduced IL-1β levels in a dose-dependent manner. The high-dose group and the Western medicine control group could significantly reduce IL-1β levels and had a good therapeutic effect (P<0.01).

[0104] Compared with the model group, the low, medium and high doses of the Chinese medicine composition of the present invention all reduced the IL-6 level (P<0.05). The Chinese medicine control group had no statistical significance, while the Western medicine control group significantly reduced the IL-6 level (P<0.01).

[0105] The above results show that the inflammation in the model group was maintained at a relatively high level, and the high-dose group of the Chinese medicine composition of the present invention can significantly reduce the levels of inflammatory factors TNF-α, IL-1β, and IL-6, with good therapeutic effect.

[0106] 4.4 Effect of Chinese herbal medicine combination on prostate pathological morphology

[0107] like Figure 4 As shown in the figure, the prostate capsule of the sham operation group was normal, with prostate glands of different sizes and regular shapes. The prostate stroma had no obvious swelling and was clear. The glands of the model group changed from columnar epithelial cells to short columns, with thicker epithelial thickness and thicker glandular duct folds. Inflammatory cell infiltration was seen between the glandular ducts, and the pathological changes were obvious (P<0.001).

[0108] Compared with the model group, all drug-treated groups showed reduced inflammatory cell infiltration, reduced epithelial wrinkles, and reduced lymphoid nodules / follicles. The high-dose group of the Chinese medicine composition of the present invention and the Chinese medicine control group showed significant changes, with significant statistical differences (P<0.001). The epithelial thickness of the Western medicine control group was significantly thinner (P<0.01); there was no significant difference in changes in inflammatory infiltration (P>0.05). 4.5 Effect of Chinese medicine composition on prostate tissue fibrosis

[0109] Masson staining can reflect the changes in the volume of collagen fibers. After staining, muscle fibers appear red and collagen fibers appear blue. Masson staining can be used to evaluate the level of prostate fibrosis in rats and further reflect the degree of inflammation in the prostate of rats.

[0110] like Figure 5As shown in the figure, the alveoli of the rats in the sham operation group were clean and mainly red muscle fibers. Compared with the control group, the blue collagen fibers of the rats in the model group increased significantly, indicating that the fibrosis of the rats in the model group was more serious (P<0.001).

[0111] Compared with the model group, the collagen fibers in the medium-dose group of the Chinese medicine composition of the present invention were reduced (P<0.05); the intimal hyperplasia in the high-dose group and the western medicine control group was alleviated, the wrinkles were smoother, and the collagen fibers were significantly reduced (P<0.001); the collagen fibers in the western medicine control group also had a significant statistical difference (p<0.01).

[0112] The above results show that the medium and high dose groups of the Chinese medicine composition of the present invention can significantly reduce the level of prostate fibrosis in rats and have a good therapeutic effect.

[0113] Example 2 Study on the therapeutic effect of the Chinese medicine composition of the present invention on chronic bacterial prostatitis

[0114] 1. Bacterial Culture

[0115] The standard pathogenic Escherichia coli ATCC25922 (Lot Number: 70027805, Beina Biotech, Beijing) was cultured in TSB (Trypticase soy broth) at 37°C for 12 hours. Before inoculation, the bacterial solution was diluted with sterile saline to a concentration of 1.0×10 6 CFU / mL suspension.

[0116] 2. Establishment of bacterial prostatitis rat model

[0117] Male SD rats, SPF grade (Beijing Weitong Lihua Experimental Animal Technology Co., Ltd., (SYXK (Tianjin) 2020-0005), weighing 250-300 g, were fed adaptively for one week and anesthetized with 10% chloral hydrate intraperitoneally, and their genital area was cleaned with iodine cotton balls. Under sterile conditions, a 1.5 cm incision was made in the middle of the lower abdomen of the rat, and the abdominal wall and peritoneum were cut to expose the bladder. The bladder was gently lifted with toothless forceps to expose the prostate. 0.05 mL of Escherichia coli (ATCC25922) suspension (1.0×10 6 CFU / ml), and the control group was injected with 0.05 mL of normal saline, and then the peritoneum and skin were sutured layer by layer and disinfected.

[0118] 3. Experimental Grouping and Drug Administration

[0119] 24 hours after modeling, the rats were randomly divided into 10 groups: sham operation group (Sham), model group (Model), low-dose group of Chinese medicine composition (Low), medium-dose group of Chinese medicine composition (Medium), high-dose group of Chinese medicine composition (High), Chinese medicine control group (Polygonum capitatum, THL), western medicine control group (ciprofloxacin, CIP), comparative example 1 group (D1), comparative example 2 group (D2), comparative example 3 group (D3), with 10 animals in each group, and the drugs were administered by gavage.

[0120] The dosage of the rats in the low-dose group of the Chinese medicine composition was 12.24 g of crude drug / kg body weight, the dosage of the rats in the medium-dose group of the Chinese medicine composition was 36.72 g of crude drug / kg body weight, the dosage of the rats in the high-dose group of the Chinese medicine composition was 110.16 g of crude drug / kg body weight, the dosage of the comparative examples 1-3 was 36.72 g of crude drug / kg body weight, the dosage of the rats in the Chinese medicine control group was 2.7 g of crude drug / kg body weight, and the dosage of the rats in the Western medicine control group was 0.045 g / kg body weight. Each drug group was given drug treatment for 7 days, and the model group and the sham operation group were given an equal volume of normal saline by gavage, once a day.

[0121] 4. Detection indicators

[0122] 4.1 Prostate Index

[0123] The prostate index is an important factor in evaluating the development of prostatitis and reflects the degree of prostate inflammation. The prostate index increases with the progression of inflammation. Rats were anesthetized with 10% chloral hydrate, the abdominal cavity was opened under sterile conditions, the prostate tissue was removed, cleaned, and weighed. The prostate visceral coefficient was calculated according to the following formula:

[0124] Prostate index = total wet weight of prostate tissue / rat body weight

[0125] 4.2 Bacterial colony culture

[0126] Each group of prostate tissue was cultured in an agar dish, and the number of colonies was detected. Under a sterile environment, the prostate tissue of the same area of ​​each rat was weighed, and the prostate tissue was chopped with sterile tissue scissors to prepare a prostate tissue homogenate. The supernatant of the tissue fluid was spread on an agar plate, 100 μL was spread on each plate, and the number of colonies was counted after culturing at 37°C for 24 hours. The bacterial count was expressed as the logarithmic mean of the total number of colonies per gram of prostate tissue (Log CFU / g of prostate tissue). 4.3 ELISA method was used to detect the expression of tissue inflammatory factors TNF-α and IL-1β

[0127] TNF-α is an important pro-inflammatory cytokine that plays an important role in the occurrence and development of chronic prostate inflammation. TNF-α can be used as an indicator of the strength of the local immune response of the prostate, the diagnosis of chronic prostatitis and the detection of the disease. Reducing the level of the pro-inflammatory cytokine TNF-α is of great significance for preventing organ damage. In bacterial prostatitis, TNF-α is usually associated with IL-1β, so the ELISA method is used to detect the expression of tissue inflammatory factors TNF-α and IL-1β. According to the instructions of the TNF-α (RA20035, Beiyinlai Biology) and IL-1β (RA20020, Beiyinlai Biology) ELISA kits, the expression of inflammatory factors TNF-α and IL-1β in tissues was detected respectively.

[0128] 4.4 Histopathological analysis

[0129] The most direct indicator of prostatitis is obvious inflammation of prostate tissue. Pathological observation can accurately determine the severity of prostatitis. Use an optical microscope to observe whether there is inflammatory cell infiltration, inflammatory cell types, tissue edema, glandular epithelial necrosis, shedding, and glandular epithelial disappearance. The prostate tissue of the same area of ​​each rat was fixed in 4% tissue fixative, dehydrated, and routinely paraffin-embedded and sectioned. The section thickness was 5μm, and H&E staining and Masson staining were performed for histological scoring and quantitative analysis. Observation under an optical microscope, the severity of prostatitis was scored according to the three parameters of chronic inflammatory cell infiltration number, acinar changes and interstitial fibrosis in representative areas, and the Masson staining results were quantitatively analyzed by Imagej software.

[0130] 4.5 Routine blood test

[0131] The infection status of rats in each group after treatment was observed. On the 8th day after modeling, blood was collected from the orbits of rats in each group into anticoagulant tubes and shaken evenly. The white blood cell count (WBC), absolute neutrophil value (NEU#), and lymphocyte ratio (LYM#) of rats were detected using a fully automatic blood and body fluid analyzer.

[0132] 4.6 Serum biochemical test

[0133] Blood was collected from the abdominal aorta of rats in each group, and the whole blood was centrifuged at 3000 rpm / min for 15 min. 100 μl of supernatant was aspirated and stored at -80°C for later use. The levels of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), urea nitrogen (BUN) and creatinine (CRE) in rats were measured by automatic biochemical analyzer to detect the safety of drugs in each group.

[0134] 4.7 Data Processing

[0135] The data were processed using SPSS 22.0 software. The data were compared among the groups using one-way analysis of variance and multiple comparisons, and the graphs were drawn using Sigma Plot 14. P < 0.05 indicated that the difference was statistically significant.

[0136] 5. Experimental Results

[0137] 5.1 Effects of Chinese herbal medicine combination on prostate index

[0138] Swelling of prostate tissue is one of the characteristics of prostatitis. Prostate index is often used as a detection indicator of CBP, which can reflect the degree of tissue inflammation to a certain extent.

[0139] The effect of the Chinese medicine composition of the present invention on the prostate index of rats with chronic bacterial prostate is shown in Table 2. Compared with the rats in the sham operation group, the prostate index of the rats in the model group was significantly increased (P<0.01), indicating that the prostate tissue had obvious inflammatory symptoms under the disease state. Compared with the rats in the model group, the prostate index of the rats in the western medicine control group was significantly reduced (P<0.001); the prostate index of the rats in the low, medium and high dose groups of the Chinese medicine composition of the present invention all showed a downward trend, and showed dose dependence, but there was no significant difference between the low dose group of the Chinese medicine composition, the Chinese medicine control group and the comparative examples 1-3 groups and the model group (P>0.05); the results showed that the medium and high dose groups of the Chinese medicine composition of the present invention can significantly improve the swelling and inflammatory state of the prostate tissue.

[0140] Table 2 Effect of Chinese medicine combination on prostate index of rats

[0141]

[0142] Compared with the sham operation group, ** p<0.01; compared with the model group, ## p<0.01, ### p<0.001.

[0143] 5.2 Effect of Chinese herbal medicine combination on bacterial counts in the prostate

[0144] Table 3 shows the effect of the Chinese medicine composition of the present invention on the bacterial count in rats with chronic bacterial prostate.

[0145] As shown in Table 3, no obvious bacterial colonies were observed in the prostate tissue of the sham-operated rats. Compared with the control group, the number of bacteria in the prostate of the model group rats increased significantly. Compared with the model group, the colony counts of the prostate tissue of the rats in the western medicine control group were significantly reduced; the colony counts of the prostate tissue of the rats in the low, medium and high dose groups of the Chinese medicine composition of the present invention were also reduced, and they were dose-dependent, with significant differences between the medium and high dose groups, and no significant differences between the comparative examples 1-3 groups; the colony counts of the prostate tissue of the rats in the Chinese medicine control group were significantly reduced.

[0146] Table 3 Effect of Chinese medicine combination on bacterial count in rat prostate

[0147]

[0148]

[0149] Compared with the sham operation group, *** p<0.001; compared with the model group, # p<0.05, ## p<0.01, ### p<0.001.

[0150] 5.3 Effects of Chinese herbal medicine combination on inflammatory factors in prostate tissue

[0151] (1) Level of inflammatory factor TNF-α

[0152] The effect of the Chinese medicine composition on inflammatory factors in rat tissues with chronic bacterial prostate is shown in Table 4 and Figure 6 .

[0153] like Figure 6 As shown, compared with the sham operation group, the level of tissue inflammatory factor TNF-α in the model group rats was significantly increased. Compared with the model group rats, the level of tissue inflammatory factor TNF-α in the ciprofloxacin group rats was significantly reduced; the TNF-α of the rats in the low, medium and high dose groups of the Chinese medicine composition of the present invention was also reduced, and it was dose-dependent, among which the high dose group had a significant difference; the TNF-α of the rats in the comparative example 1-3 groups and the Chinese medicine control group was not significantly reduced.

[0154] Table 4 Effect of Chinese medicine combination on inflammatory factor TNF-α in rat prostate tissue

[0155]

[0156] Compared with the sham operation group, *** p<0.001; compared with the model group, # p<0.05, ### p<0.001.

[0157] (2) Levels of inflammatory factor IL-1β

[0158] The effect of the Chinese medicine composition of the present invention on inflammatory factors in rat tissues with chronic bacterial prostate. The results are shown in Tables 5 and Figure 7 .

[0159] like Figure 7As shown in the figure, compared with the sham operation group, the level of tissue inflammatory factor IL-1β in rats in the model group was significantly increased.

[0160] Compared with the rats in the model group, the level of tissue inflammatory factor IL-1β in the rats in the western medicine control group was significantly reduced; the IL-1β in the rats in the low, medium and high dose groups of the Chinese medicine combination was also reduced, and was dose-dependent, with significant differences in the high dose group; the IL-1β in the rats in the Chinese medicine control group and comparison groups 1-3 was not significantly reduced.

[0161] Table 5 Effect of Chinese medicine combination on inflammatory factor IL-1β in rat prostate tissue

[0162]

[0163] Compared with the sham operation group, *** p<0.001; compared with the model group, # p<0.05, ### p<0.001.

[0164] 5.4 Effects of Chinese herbal medicine combination on prostate histopathology

[0165] (1) The results of the H&E staining test are shown in Figure 8 .

[0166] Sham group: The prostate epithelial cells of rats were arranged neatly, smooth and flat, without obvious hyperplasia, the secretions in the cavity were pink, and there were very few inflammatory cells in the interstitium.

[0167] Model group: The number of prostate glands in rats was significantly reduced, the glandular epithelial tissue was destroyed to varying degrees, the glandular epithelium fell off, fused or thinned into a single layer, the glandular cavity disappeared, and a large number of inflammatory cells infiltrated.

[0168] Low-dose group (Low) of the Chinese medicinal composition of the present invention: the number of acini is small, the acini are dilated to varying degrees, the interstitial tissue is significantly widened, there is a small amount of secretion in the acini, and more inflammatory cell infiltration can be seen.

[0169] In the medium dose group (Medium) of the Chinese medicine composition of the present invention, a certain degree of hyperplasia of the glandular epithelium and a certain degree of fibrosis in the stroma were observed. The infiltration of inflammatory cells was greatly reduced compared with the model group, but scattered distribution was still observed.

[0170] Traditional Chinese medicine control group (THL): The glandular epithelial tissue was still damaged to varying degrees, with more acini than in the model group, but the glandular epithelium became thinner into a single layer and infiltrated with inflammatory cells.

[0171] The high-dose group (High) of the Chinese medicine composition of the present invention and the ciprofloxacin group (CIP): there was no obvious hyperplasia of the glandular epithelium, the alveolar structure was basically normal, the morphology was relatively flat and regular, there was no obvious edema in the interstitium, a small number of inflammatory cells were distributed therein, there was no obvious fibrosis, and the degree of chronic inflammation was greatly reduced compared with the model group and others.

[0172] (2) The effect of the Chinese medicine composition of the present invention on the collagen volume fraction of rat tissue with chronic bacterial prostate. The results are shown in Table 6. Fig. 9 and Fig.10 .

[0173] like Fig. 9 and Fig.10 As shown in the figure, compared with the sham operation group, the tissue collagen volume fraction of the rats in the model group was significantly increased.

[0174] Compared with the rats in the model group, the tissue collagen volume fraction of the rats in the western medicine control group was significantly reduced; the collagen volume fractions of the low, medium and high dose groups of the Chinese medicine composition of the present invention were also reduced, among which the high dose group had a significant difference; the collagen volume fraction of the rats in the Chinese medicine control group was significantly reduced.

[0175] Table 6 Effect of the Chinese medicine composition of the present invention on the collagen volume fraction of rat prostate tissue

[0176]

[0177] Note: Compared with the sham operation group, *** p<0.001; compared with the model group, # p<0.05, ### p<0.001.

[0178] 5.5 Effect of the Chinese medicinal composition of the present invention on the number of white blood cells, neutrophils and lymphocytes in rat blood routine test

[0179] The effect of the Chinese medicine composition on the number of white blood cells, neutrophils and lymphocytes in the blood routine test of rats with chronic bacterial prostate is shown in Table 7.

[0180] (1) Compared with the sham operation group, the number of leukocytes in the blood of the model group rats was significantly increased. Compared with the model group rats, the number of leukocytes in the blood of the western medicine control group rats was significantly reduced; the number of leukocytes in the blood of the rats in the low, medium and high dose groups of the Chinese medicine composition of the present invention was also reduced, and it was dose-dependent, and the high dose group had a significant difference; the number of leukocytes in the blood of the rats in the Chinese medicine control group was significantly reduced, and there was no significant difference between the comparative examples 1-3 and the model group.

[0181] (2) Compared with the sham operation group, the absolute value of neutrophils in the blood of the model group rats was significantly increased. Compared with the model group rats, the absolute value of neutrophils in the blood of the western medicine control group rats was significantly reduced; the absolute value of neutrophils in the blood of the rats in the low and medium dose groups of the Chinese medicine composition of the present invention was also significantly reduced, and showed a dose-dependent manner, with significant differences between the medium and low dose groups, and no downward trend in the high dose group and the comparative examples 1-3 groups; the absolute value of neutrophils in the blood of the rats in the Chinese medicine control group had a downward trend but no significant difference.

[0182] (3) Compared with the sham operation group, the proportion of lymphocytes in the blood of the model group rats was significantly increased. Compared with the model group rats, the proportion of lymphocytes in the blood of the western medicine control group rats was significantly reduced; the proportion of lymphocytes in the blood of the rats in the low, medium and high dose groups of the Chinese medicine composition of the present invention was also reduced, and there was a significant difference; the proportion of lymphocytes in the blood of the rats in the Chinese medicine control group was significantly reduced, and there was no significant difference in the lymphocyte proportion of the comparative example 1 group and the comparative examples 2-3 groups.

[0183] Table 6 Effects of Chinese herbal medicine combination on the number of white blood cells, neutrophils and lymphocytes in rat blood routine

[0184]

[0185] Compared with the sham operation group, * p<0.05; compared with the model group, # p<0.05, ## p<0.01, ### p<0.001.

[0186] 5.6 Effects of Chinese herbal medicine combination on serum biochemistry in rats

[0187] The effect of the Chinese medicine composition on serum biochemistry of CBP rats is shown in Table 7. Compared with the sham operation group, the serum biochemical indexes ALT, AST, BUN, and CRE of the model group rats were not significantly reduced or increased. Compared with the model group rats, the indexes of the other drug administration groups were not significantly reduced or increased, proving that the drugs at each dose had no obvious liver and kidney toxicity and were highly safe.

[0188] Table 7 Effects of Chinese medicine combination on serum biochemical indexes in rats

[0189]

[0190]

Claims

1. A Chinese medicine composition for treating prostate disease, characterized in that: The traditional Chinese medicine composition is mainly prepared from the following raw materials: 3-15 parts by weight of cassia twig, 20-40 parts by weight of oyster, 20-40 parts by weight of dragon bone, 10-30 parts by weight of radix polygoni multiflori, 10-30 parts by weight of pyrrosia, and 20-40 parts by weight of salvia miltiorrhiza.

2. The Chinese medicine composition according to claim 1, characterized in that The traditional Chinese medicine composition is prepared from the following raw materials: 15 parts by weight of cassia twig, 40 parts by weight of oyster, 40 parts by weight of dragon bone, 30 parts by weight of radix polygoni multiflori, 30 parts by weight of pyrrosia and 40 parts by weight of salvia miltiorrhiza.

3. The Chinese medicine composition according to claim 2, characterized in that: The traditional Chinese medicine composition is prepared from the following raw materials: 6 parts by weight of cassia twig, 30 parts by weight of oyster, 30 parts by weight of dragon bone, 20 parts by weight of radix polygoni multiflori, 20 parts by weight of pyrrosia and 30 parts by weight of salvia miltiorrhiza.

4. The Chinese medicine composition according to any one of claims 1 to 3, characterized in that: The prostate disease is one or more of prostatitis, prostate hyperplasia and prostate cyst.

5. The Chinese medicine composition according to claim 4, characterized in that: The prostatitis is one or more of acute bacterial prostatitis, chronic bacterial prostatitis, chronic prostatitis / chronic pelvic pain syndrome and asymptomatic prostatitis.

6. The Chinese medicine composition according to claim 4, characterized in that: The prostatitis is divided into damp-heat descending type, damp-heat stasis type, qi stagnation and blood stasis type, yin deficiency and hyperactivity of fire type, kidney yang deficiency and fatigue type or spleen deficiency and dampness type according to TCM syndrome differentiation.

7. The Chinese medicine composition according to any one of claims 1 to 6, characterized in that: The Chinese medicine composition is prepared by the following method: (1) calcining dragon bones and oysters, crushing and mixing to obtain a mixed powder, sieving, and subjecting the mixed powder to decoction extraction or reflux extraction to obtain an extract; (2) extracting Danshen with 40-95% ethanol to obtain an ethanol extract; (3) extracting Radix Dioscoreae and Pyrola with water to obtain a water extract; (4) filtering the extract of step (3), concentrating under reduced pressure, adding ethanol and filtering to obtain a filtrate; (5) Mix the extracts of steps (1), (2) and (4) to obtain.

8. The Chinese medicine composition according to claim 7, characterized in that: The Chinese medicine composition is prepared by the following method: (1) The dragon bone and oyster are crushed and mixed to obtain a mixed powder, and the mixed powder is sieved. The mixed powder is mixed with water at a ratio of 1:10, and the mixture is boiled at 95-100° C. for 1 hour. After filtering, the decoction is considered to have completed one extraction. The extraction is repeated 3-4 times and the decoction is combined; (2) extracting Danshen with 50-80% ethanol to obtain an ethanol extract; (3) extracting Radix Dioscoreae and Pyrola with water to obtain a water extract; (4) filtering the water extract of step (3), concentrating under reduced pressure to a relative density of 1.05 to 1.25 at 50° C., adding ethanol to make the alcohol content reach more than 40%, and filtering to obtain a filtrate; (5) The extracts of steps (1), (2) and (4) are mixed to obtain the product.

9. The Chinese medicine composition according to any one of claims 1 to 6, characterized in that: The Chinese medicine composition is prepared into a Chinese medicine oral preparation.

10. The Chinese medicine composition according to claim 9, characterized in that: The oral preparation is one or more of pills, granules, tablets, dripping pills, micropills and capsules.

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