CN116785328A - Application of Inonotus obliquus extract in anti-prostatic hyperplasia and chronic prostatitis - Google Patents

Abstract

The invention discloses the application of Inonotus obliquus extract in preparing medicines for preventing and/or treating prostatic hyperplasia and chronic prostatitis. Animal experiments show that the extract has anti-inflammatory, antioxidant, anti-fibrosis and other activities, and can It is used in the prevention and/or treatment of prostatic hyperplasia and chronic prostatitis in the field of pharmaceuticals and health care products.

Description

Application of Inonotus obliquus extract in anti-prostatic hyperplasia and chronic prostatitis

Technical field

The invention belongs to the technical field of natural medicine extract application, and particularly relates to the application of Inonotus obliquus extract in resisting prostatic hyperplasia and prostatitis.

Background technique

As the aging of the population becomes more and more serious, benign prostatic hyperplasia (BPH) has gradually become one of the most common and harmful diseases for elderly men. Benign prostatic hyperplasia affects nearly half of men aged around 50 and nearly 80% of men aged 70 and older. Benign prostatic hyperplasia is a chronic disease characterized by abnormal proliferation of stromal and epithelial cells in the hyperprostatic zone, resulting in urethral obstruction. The possible mechanisms of prostatic hyperplasia are related to aging, chronic inflammation, obesity, oxidative stress, etc., but they are largely unknown, and there are no safe and effective treatments.

Prostatitis is a common and frequently-occurring disease in urology. Common treatment methods such as antibiotics, anti-inflammatory drugs, hormones, etc. are difficult to achieve satisfactory clinical results.

Inonotus obliquus is a medicinal and edible fungus, also called Chaga. It is a fungus of the genus Inonotus of the family Dermatomycetes and has the effect of nourishing qi and nourishing yin. The sclerotia of Inonotus obliquus is a tuber-shaped sterile fruiting body with a surface that is yellowish brown to black, hard and uneven, and the interior is yellow after being broken. Inonotus obliquus mainly lives under the bark of white birch, silver birch, elm, alder, etc. Its growth areas are mainly distributed in cold areas of 40 to 50 degrees north latitude in the northern hemisphere.

Inonotus obliquus mainly contains chemical components such as polysaccharides, polyphenols, triterpenes, alkaloids, and melanin. Modern pharmacological research shows that Inonotus obliquus has various biological activities such as anti-tumor, hypoglycemic, immune regulation, anti-inflammatory, and antioxidant.

However, there have been no reports on the anti-prostatic hyperplasia and prostatitis effects of Inonotus obliquus.

Contents of the invention

The object of the present invention is to provide the application of Inonotus obliquus extract in resisting prostatic hyperplasia and chronic prostatitis.

Specifically, the present invention relates to the use of Inonotus obliquus extract as a pharmaceutical active ingredient in the preparation of medicines for preventing and/or treating prostatic hyperplasia.

Further, the present invention also relates to the use of Inonotus obliquus extract as a pharmaceutical active ingredient in the preparation of medicines for preventing and/or treating prostatitis.

Furthermore, the present invention relates to the use of Inonotus obliquus extract as a pharmaceutical active ingredient in the preparation of medicines for preventing and/or treating chronic prostatitis.

The Inonotus obliquus extract of the present invention is a polysaccharide and glycoprotein active ingredient extracted from Inonotus obliquus by water extraction and alcohol precipitation.

Specifically, the Inonotus obliquus extract of the present invention is extracted from Inonotus obliquus according to the following method.

Add water to Inonotus obliquus according to the material-to-liquid ratio of 1:20 and infiltrate it for 0.5 to 1 hour, heat and reflux for extraction for 1.5 to 2.5 hours, filter, collect the filtrate, and extract three times in total. Combine the filtrate, concentrate to a density of 1.05~1.10 (50°C), add 95% (V/V) ethanol until the ethanol concentration in the concentrated solution reaches 65~80% (V/V), let it stand at room temperature for 12 hours, and centrifuge to collect the precipitate. , dissolved in water and filtered, then freeze-dried to prepare Inonotus obliquus extract.

Further, the present invention uses the Inonotus obliquus extract as a pharmaceutical active ingredient, adds a pharmaceutically acceptable pharmaceutical carrier or excipient that can be used in combination with the active ingredient, and prepares it into a powder or granule. , tablets, pills, capsules and other common oral dosage forms in pharmacy.

Furthermore, the present invention can also use the Inonotus obliquus extract as a functional additive and add it to various foods to make functional foods. The foods include but are not limited to steamed buns, bread, cakes, biscuits or instant noodles. wait.

The present invention constructs a mouse model of prostatic hyperplasia induced by testosterone propionate (TP) and administers different doses of Inonotus obliquus extract by gavage, proving that the Inonotus obliquus extract has obvious effects on preventing and/or treating prostatic hyperplasia. As a result, the prostate wet weight index of the model mice decreased significantly, the ACP activity and type II 5-α reductase content in the mouse serum decreased, the SOD and CAT activities in the prostate tissue increased, and the GSH content levels tended to normalize.

At the same time, rats with chronic prostatitis were given different doses of Inonotus obliquus extract, which showed that the extract of Inonotus obliquus could better improve the pathological damage of prostate tissue in rats with chronic prostatitis and play a protective role in prostate tissue. It also verified and revealed its mechanism of reducing symptoms such as inflammation and pain by reducing the content of COX-2 and PGE2 in rat prostate tissue.

Therefore, Inonotus obliquus extract can be used in the field of pharmaceuticals and health products to prevent and/or treat prostatic hyperplasia and chronic prostatitis.

Detailed ways

Specific embodiments of the present invention will be described in further detail below with reference to examples. The following examples are only used to illustrate the technical solution of the present invention more clearly, so that those skilled in the art can better understand and utilize the present invention, but are not intended to limit the protection scope of the present invention.

The names and abbreviations of the experimental methods, production processes, instruments and equipment involved in the embodiments of the present invention are conventional names in the field, and are very clear and unambiguous in the relevant fields of use. Those skilled in the art can use these names to Understand common process steps and apply appropriate equipment, performing under common conditions or conditions recommended by the manufacturer.

There are no special restrictions on the source of various raw materials or reagents used in the embodiments of the present invention, and they are all conventional products that can be purchased on the market. It can also be prepared according to conventional methods well known to those skilled in the art.

In the present invention, the total polysaccharide content of the Inonotus obliquus extract is measured using the phenol-sulfuric acid method, the total polyphenol content is measured using the Folin-phenol method, and the total saponin content is measured using the vanillin-glacial acetic acid-perchloric acid spectrophotometric method. , Total protein content was measured using the Coomassie brilliant blue method.

Example 1: Preparation of Inonotus obliquus extract.

Take the sclerotia of Inonotus obliquus, crush it with a traditional Chinese medicine crusher, and then pass it through a 60-mesh sieve to obtain bacterial powder for later use.

Weigh 1000g of dried Inonotus obliquus powder, add distilled water at a material-to-liquid ratio of 1:20, soak for 1 hour, heat to 80°C and reflux for 1.5 hours, filter, and collect the filtrate. Extract 3 times under the same conditions.

The filtrate obtained from three extractions was combined and concentrated at 60°C to a density of 1.05 to 1.10. After cooling, add 95% ethanol to the concentrated solution until the final ethanol volume fraction is 80%, and leave it for 12 hours for precipitation.

Then centrifuge at 3000 r/min for 15 min, collect the precipitate, dissolve it in distilled water and filter it, collect the filtrate and freeze-dry it to obtain Inonotus obliquus extract.

After measurement, the active ingredients contained in the Inonotus obliquus extract obtained above were: polysaccharide 64.2%, protein 9.6%, polyphenol 10.8%, and saponin 8.6%.

Example 2: Animal test on anti-prostatic hyperplasia of Inonotus obliquus extract.

Kunming mice were randomly divided into blank group, model control group, Inonotus obliquus extract high, medium and low dose groups, and finasteride positive control group, a total of 6 groups, with 10 mice in each group.

The model control group was administrated with normal saline and subcutaneously injected with 5 mg/kg testosterone propionate (TP) every day.

The mice in the blank group were intragastrically administered with physiological saline and subcutaneously injected with the same volume of olive oil every day.

In the low-dose group of Inonotus obliquus extract, 200 mg/kg of Inonotus obliquus extract was administered intragastrically every day, and 5 mg/kg of testosterone propionate (TP) was injected subcutaneously.

The medium-dose group of Inonotus obliquus extract was given 400 mg/kg of Inonotus obliquus extract by intragastric administration every day, and 5 mg/kg of testosterone propionate (TP) was injected subcutaneously.

In the high-dose Inonotus obliquus extract group, 800 mg/kg of Inonotus obliquus extract was administered intragastrically every day, and 5 mg/kg of testosterone propionate (TP) was injected subcutaneously.

The finasteride positive control group was given 1 mg/kg finasteride solution by intragastric administration and subcutaneous injection of 5 mg/kg testosterone propionate (TP) every day.

The administration lasted for 3 weeks, and the administration volume was 10 ml/kg, and each intragastric administration was performed 1 hour before the subcutaneous injection of TP. The mice were allowed to eat and drink 24 hours after the last injection of TP.

Blood was collected from mice in each group through the orbit, left for 30 minutes, and centrifuged at 3000r for 15 minutes. The supernatant was collected and stored at -80°C for detection of ACP and type II 5-alpha reductase levels in the serum.

The mice were sacrificed by cervical dislocation, the prostate was quickly removed, and its wet weight was measured to calculate the prostate index (PI).

Part of the ventral lobe of the prostate was taken out to make 10% tissue homogenate, which was used to detect the MDA and GSH contents, as well as SOD and CAT activities in the prostate tissue.

All data are expressed as mean ± standard deviation, and one-way analysis of variance was used to test the significance of data differences between multiple groups.

Compared with the model control group, *, P <0.05; **, P <0.01.

The data in Table 1 show that compared with the blank group, the ACP activity in the serum of mice induced by TP increased significantly, and the ACP activity of mice treated with the positive drug finasteride decreased significantly. At the same time, the high, medium and low doses of Inonotus obliquus extract can also reduce the activity of ACP. The level of type II 5-alpha reductase in the serum of the model control group increased significantly, while the level of type II 5-alpha reductase in mice treated with Inonotus obliquus extract and finasteride decreased to varying degrees.

Compared with the model control group, *, P <0.05; **, P <0.01.

Table 2 shows that compared with the blank group, the weight of mice induced by TP increased significantly, with obvious changes. At the same time, the study also found that the prostate wet weight index of mice in the high, medium and finasteride positive control groups of Inonotus obliquus extract was significantly higher than that of the model control group during 3 consecutive weeks of treatment. decline.

Compared with the blank group, *, P <0.05; compared with the model control group, #, P <0.05.

The data results in Table 3 show that treating TP-induced prostatic hyperplasia mice with high, medium and low dose groups of Inonotus obliquus extract can significantly increase their SOD and CAT activities and normalize GSH levels. In addition, the application of Inonotus obliquus extract also inhibited the process of lipid peroxidation. These evidences indicate that Inonotus obliquus extract has significant antioxidant activity.

Example 3: Animal test on the anti-prostatitis effect of Inonotus obliquus extract.

SD rats were randomly divided into blank group, model control group, Inonotus obliquus extract high, medium and low dose groups, Qianliekang positive control group, a total of 6 groups, 10 rats in each group.

Except for the blank group, rats in the other groups were subjected to modeling. On days 1, 14, and 28 of the experiment, a mixture (prepared Wistar rat prostate protein purification solution with a concentration of 1 mg/ml added to the same volume of complete Freund's adjuvant) was injected into different subcutaneous parts of the rats. (obtained by post-emulsification of CFA)). The injection sites are: subcutaneous on the left foot pad, subcutaneous on the right foot pad, subcutaneous on the base of the tail, subcutaneous on the left scapula, and subcutaneous on the right scapula. Inject 0.2ml at each point.

After successful modeling, the blank group and model control group were administered 10ml/kg normal saline; the high, medium and low dose groups of Inonotus obliquus extract were administered 800mg/kg, 400mg/kg and 200mg/kg; Qianliekang positive The control group was given 5g/kg Qianliekang by gavage. Once a day for 10 consecutive days.

Ten days after intragastric administration, blood was collected from the rats in each group via the abdominal aorta after anesthesia. Whole blood was collected in a container containing heparin, placed for 30 minutes, and centrifuged at 2500r for 30 minutes. The supernatant was collected and stored at -80°C for subsequent experimental inspection and identification.

The anterior lobe of the rat prostate (AP) and the ventral lobe of the prostate (VP) were removed, and part of them was fixed in 10% neutral formalin solution, dehydrated with ethanol, embedded in paraffin, and sectioned into 5 μm sections for hematoxylin-eosin analysis. dyeing. Observe the slices and rate the degree of prostate inflammation. The other part was made into tissue homogenate, centrifuged at 3000r for 20 minutes, the supernatant was taken out, and the contents of IL-1β, COX-2, PGE2, and β-EP in the prostate tissue homogenate were measured by ELISA.

All data are expressed as mean ± standard deviation, and one-way analysis of variance was used to test the significance of data differences between multiple groups.

The pathological inflammation grading of prostate tissue refers to the international prostatitis histological diagnosis and grading standards: Grade I refers to scattered inflammatory cells, the number of inflammatory cells is 1 to 10/HP, and is recorded as +; Grade II refers to the accumulation of inflammatory cells without destruction of glandular epithelial tissue. Or lymphoid nodules/follicles are formed, the number of inflammatory cells is 11-20/HP, scored as ++; Grade III refers to the accumulation of inflammatory cells, partial gland epithelial tissue destruction or the formation of lymphoid nodules/follicles, the number of inflammatory cells is >20 per HP, recorded as +++; grade VI refers to the accumulation of inflammatory cells, destruction of a large amount of glandular epithelial tissue or formation of lymphoid nodules/follicles, and the number of inflammatory cells filling the field of view/HP, recorded as ++++.

Compared with the blank group, ##, P <0.01; compared with the model control group, *, P <0.05; **, P <0.01.

The results in Table 4 show that the differences between the high, medium and low dose groups of Inonotus obliquus extract and the Liekang positive control group are statistically significant compared with the model control group; the differences between the low dose group of Inonotus obliquus extract and the blank group are all statistically significant. , the difference is statistically significant, and the differences between the other groups and the blank group are not statistically significant. It shows that the high, medium and low dose groups of Inonotus obliquus extract and the Qianliekang positive control group can better improve the pathological damage of prostate tissue in rats with chronic prostatitis and play a protective role in prostate tissue.

Compared with the blank group, *, P <0.05; **, P <0.01; compared with the model control group, #, P <0.05;##, P <0.01.

Table 5 shows that IL-1β in the prostate tissue homogenate of rats in the model control group was significantly higher than that in the blank group, consistent with its pathological manifestations, indicating that there is a strong inflammatory response in the prostate tissue of rats with chronic prostatitis, and This inflammatory response can be induced by an autoimmune response.

Inonotus obliquus extract can significantly reduce the level of IL-1β in the prostate tissue of rats with experimental chronic prostatitis, so it has a good anti-inflammatory effect, and the high-dose group has the best effect. Since IL-1β also promotes fibrosis, Inonotus obliquus extract can not only reduce IL-1β content and reduce inflammatory response, but also reduce the proliferation of fibrous connective tissue and play a certain protective role in prostate tissue. .

In addition, the contents of COX-2 and PGE2 in the prostate tissue of rats in the model control group were significantly increased compared with the blank group, suggesting that COX-2 and PGE2 are an important link in the pathogenesis of chronic prostatitis. Inonotus obliquus extract can significantly reduce the COX-2 and PGE2 contents in the prostate tissue of rats in the model control group, thereby reducing symptoms such as inflammatory response and pain. Among them, the high-dose group has the best efficacy, and its efficacy is not significantly different from that of the Qianliekang positive control group. At the same time, Inonotus obliquus extract can significantly increase the β-EP content in prostate tissue, thereby enhancing the local analgesic effect and reducing clinical symptoms.

The above embodiments of the present invention do not describe all the details in detail, nor do they limit the present invention to only the above embodiments. Various changes, modifications, substitutions and modifications made to these embodiments by those of ordinary skill in the art without departing from the principle and purpose of the invention should be included in the protection scope of the invention.

Claims (3)

1. Application of Inonotus obliquus extract in preparing medicine for preventing and/or treating prostatic hyperplasia is provided.

2. Application of Inonotus obliquus extract in preparing medicine for preventing and/or treating prostatitis is provided.

3. The application of Inonotus obliquus extract in preparing medicine for preventing and/or treating chronic prostatitis is provided.