CN101254223A - New use of pericarpium Granati and semen Granati extract for improving prostate symptom - Google Patents

Abstract

The invention provides a method for preparing a medicine for resisting prostatic hyperplasia and prostatitis by taking pomegranate (Punica granatum Linn) peel and pomegranate seed extracts as raw materials, and also can be used for preparing health food and food additives for resisting prostatic hyperplasia and prostatitis. Experimental results show that the pomegranate bark extract and the pomegranate seed extract can effectively improve the weight and the prostate index of a prostate gland of a mouse with benign prostatic hyperplasia, increase the level of Malondialdehyde (MDA) and the level of testosterone (testosterone) in serum in the prostate gland, improve the antioxidant capacity in plasma and regulate the level of Nitric Oxide (NO) in the plasma. Furthermore, the extract of pericarpium Granati and semen Granati can also improve the density of lecithin corpuscle and leukocyte count of prostate of rats with prostatitis, and the activity of acid phosphatase (PACP) in blood plasma, and can regulate Malondialdehyde (MDA) and NO levels in prostate. In vitro experiments, it can be observed that the pomegranate peel extract and the pomegranate seed extract have strong antioxidation. The pomegranate peel extract and the pomegranate seed extract have the characteristics of high safety, exact biological activity and the like. Therefore, the usage amount of the pomegranate bark extract and the pomegranate seed extract in various preparation finished products of the medicine or the health food and the food additive can be 0.001-100% by weight.

Description

Novel use of pomegranate peel and pomegranate seed extracts to improve prostate symptoms

technical field

The present invention relates to the field of medicine, in particular to the field of natural plant extracts and applications thereof, in particular to the use of pomegranate (Punicagranatum Linn.) peel and pomegranate seed extracts for the preparation of anti-prostatic hyperplasia and anti-prostatitis medicines, health food and food additives Applications.

Background technique

Prostatic hyperplasia, commonly known as prostatic hypertrophy, refers to the enlargement of the prostate organ due to the increase in the number of prostate parenchymal cells. Prostatic hyperplasia is one of the common diseases of elderly men. The pathological changes of benign prostatic hyperplasia usually begin after the age of 40, and related symptoms appear after the age of 50. The incidence of benign prostatic hyperplasia increases with age, but the clinical symptoms are most obvious at the age of 60-70. The main symptoms of benign prostatic hyperplasia include dysuria, frequent urination, slow start of urination, weak urinary stream, intermittent or dribbling, etc. What’s more, acute urinary retention and filling incontinence can occur, which seriously affect the quality of life of elderly men, and even Various complications occur, threatening the life of the patient, and become one of the most important diseases affecting the health of elderly men in my country. It is generally believed that the mechanism of benign prostatic hyperplasia is related to the accumulation of dihydrotestosterone in the gland in elderly men.

Prostatitis is a frequently-occurring and common disease of the male urinary system. It is not a single disease. Its etiology can be primary or secondary to tissues and organs, and it is also related to infection and inflammation. Among them, acute prostatitis is mostly an infectious disease, while chronic prostatitis is regarded as a clinical symptom characterized by abnormal urination and other symptoms formed under the action of infection or non-infection factors. Since the etiology of non-infectious prostatitis is not clear, it can be considered that there are various factors causing prostatitis and the mechanism of action is complex. In recent years, people have paid great attention to the relationship between genetic factors, endocrine disorders, aging and the onset of prostatitis. They have also noticed that the physical and mental fatigue caused by stress load and the long-term congestion of the prostate caused by specific work may directly participate in or affect the development of prostatitis. occur. Many studies also believe that the lack of trace element Zn and the damage to tissues and organs caused by the body's oxidative stress state may also be one of the pathogenesis of prostatitis.

At present, the drugs for the treatment of benign prostatic hyperplasia are mainly divided into 5α-reductase receptor inhibitors, α1-adrenergic receptor blockers, anti-androgens such as slow regression tumors, and luteinizing hormone-releasing hormone drugs such as megestrol acetate. . These medicines are subject to a certain limit in the clinical dose, and the effect is also mostly limited to temporary relief, and cannot fundamentally achieve the therapeutic purpose. In addition, 5α-reductase inhibitors clinically show side effects related to sex, and α-receptor blockers are often accompanied by cardiovascular side effects, and the cost is relatively high. Anti-androgen and luteinizing hormone-releasing hormone drugs have large side effects and are rarely used. The use of antibacterial drugs in patients with chronic prostatitis can easily lead to a decrease in the immune system of flora in the body, and most antibacterial drugs cannot reach effective therapeutic levels in the prostate tissue and acinus. Studies so far have proved that the occurrence of diseases such as inflammation, tumors and aging is closely related to the excessive production of free radicals in the body or the decline in the ability to scavenge free radicals. For this reason, we are committed to finding and developing new anti-prostatic hyperplasia and anti-prostatitis drugs, health food, food and food additives from natural products of anti-active oxygen free radicals. We noticed and confirmed the anti-prostatic hyperplasia and anti-prostatitis effect of pomegranate peel and pomegranate seed extract in our research, aiming to develop its application value.

Pomegranate (Punica granatum Linn.) belongs to the pomegranate (Punicaceae) pomegranate (Punica) deciduous shrubs or small trees. It is native to the Balkan Peninsula to Iran and its adjacent areas. It has been distributed in tropical and subtropical regions of the world. Widely planted. China has a long history of introducing pomegranates and is widely cultivated. In recent years, people have paid great attention to the research on the physiological activities of natural products, especially the biological functions of traditional Chinese herbal medicines and natural plants. Modern medical research believes that pomegranate peel contains "pomegranate rhizoids", which can inhibit various bacilli and skin bacteria. Pomegranate peel contains more tannin, which has astringent and bactericidal effect, can cure diarrhea and dysentery, and can appetize and help digestion. Pomegranate peel is rich in catechins and other polyphenolic compounds, which have strong antioxidant effects such as scavenging oxygen free radicals. Pomegranate seed oil is rich in punicic acid, which has strong antioxidant capacity, can inhibit lipoxygenase, cyclooxygenase and key enzymes in the arachidonic acid pathway, and resist human inflammation and damage from oxygen free radicals. Pomegranate peel and pomegranate seed extracts have certain anti-inflammatory, antibacterial, anti-tumor effects, etc., which have been recognized by people, but there is no application discussion on the anti-prostatic hyperplasia and anti-prostatitis of pomegranate peel and pomegranate seed extracts.

Contents of the invention

The purpose of the present invention is to provide a new application of pomegranate peel and pomegranate seed extract.

The invention uses pomegranate peel and pomegranate seed extracts as raw materials to prepare medicines for resisting benign prostatic hyperplasia and prostatitis and alleviating their clinical symptoms. The prepared medicine can be taken for a long time, has high safety and has no toxic and side effects.

The extracts of pomegranate peel and pomegranate seeds described in the invention are used as raw materials for preparing food and food additives for anti-prostatic hyperplasia and anti-prostatitis and alleviating their clinical symptoms. The prepared food and food additives can be taken for a long time, have high safety and no toxic and side effects. In its application products, the usage amount of the pomegranate peel and pomegranate seed extract can account for 0.001-100% by weight.

The pomegranate peel and pomegranate seed extracts of the present invention are extracted by using water or any mixed solvent of one or two or more polar solvents such as water and lower alcohols such as methanol and ethanol, and acetone. Because pomegranate peel and pomegranate seed extracts are rich in flavonoids, polyphenols, amino acids, fatty acids, polysaccharides, and trace elements, all of which are medium-to-high polar compounds. Generally, water or different concentrations of lower alcohols, Acetone and other polar solvents or any mixture of solvents for extraction. During the extraction, the concentration of alcohol can be properly adjusted to improve the recovery rate of the active components in the pomegranate peel and pomegranate seed extract. During extraction, the content of active ingredients in the extract can be affected by changes in conditions such as temperature, method, and time. However, for the extraction of the active components in the pomegranate peel and pomegranate seed extract, the above extraction methods and any combination thereof can achieve the recovery of the extract described in the present invention.

In view of the fact that pomegranate peel and pomegranate seed extracts are developed as final products of medicine, health food and food additives, it is advisable to choose hydrous ethanol as the extraction solvent from the perspective of safety. The ratio of pomegranate peels and pomegranate seeds to the solvent is not specifically limited, but it is generally considered that the ratio of pomegranate peels and pomegranate seeds to the solvent is in the range of 1:1-50. The present invention has no specific limitation on the extraction temperature, which is applicable from room temperature to the boiling point of the solvent. Of course, extraction at room temperature, normal pressure and within the boiling point range of the solvent is ideal. Extraction time from 30 seconds to 24 hours can achieve the purpose of recovery of active ingredients to a certain extent. When extracting the pomegranate peel extract and pomegranate seed extract, substances such as sodium bicarbonate and sodium ascorbic acid can also be added to the solvent as needed. These extraction methods will not affect the biological activity of pomegranate peel and pomegranate seed extract.

The solid dosage of the pomegranate peel and pomegranate seed extract provided by the invention is generally 0.001% to 100% (W/W), and the concentration of the oral dosage form is usually in the range of 0.1% to 50% (W/W). As food additives, pomegranate peel extract and pomegranate seed extract may feel bitter and greasy as the amount added increases. Its solution can change dosage form according to market demand and consumer's psychological requirement, for example use dosage forms such as tablet, granule, capsule.

The pomegranate peel and pomegranate seed extract and its active ingredients provided by the invention can be put on the market in various oral dosage forms for anti-prostatic hyperplasia and anti-prostatitis and for relieving their clinical symptoms. Of course, the pomegranate peel and pomegranate seed extracts can also be combined with commonly used excipients and other medical carriers to make preparations. If necessary, preparation additives such as some preservatives, antioxidants, pigments, and sweeteners can also be added. More suitable excipients of pomegranate peel and pomegranate seed extract generally include lactose, sucrose, mannitol, starch, crystalline cellulose and the like. Lubricants should generally use magnesium stearate, calcium stearate, talcum powder, etc. As a solvent, purified water, ethanol, propylene glycol, etc. are usually used. Preservatives generally use chlorobutanol, benzyl alcohol, diglycolic acid, etc. Antioxidants are usually more suitable such as sulfites and ascorbic acid.

The pomegranate peel and pomegranate seed extract provided by the present invention can be added into preparations in solution or powder for oral administration. Pomegranate peel and pomegranate seed extract can also be combined with other beverage raw materials to make certain beverage products, such as pomegranate peel and pomegranate seed extract beverages, carbonated drinks, fruit juice drinks, lactic acid bacteria drinks, sports drinks, soybean milk, etc. Pomegranate peel and pomegranate seed extract can also be considered to make biscuits, chocolate, candy, chewing gum, fast food snacks, jelly snacks, bread, tofu, yogurt and other daily foods.

The pomegranate peel and pomegranate seed extract and the active components contained in the present invention can be used as medicine, health food, food or food additives for anti-prostatic hyperplasia and anti-prostatitis and for relieving their clinical symptoms. In the present invention, the doses of the pomegranate peel and pomegranate seed extracts can be appropriately selected according to the relative health status, age, gender and body weight of the user. The average adult body weight is calculated on the basis of 60 kg, and the daily oral administration of pomegranate peel and pomegranate seed extract can generally be considered within the range of 1 g to 5 g, divided into doses. Of course, there are no problems such as side effects if there are more than 5 grams.

In the present invention, prostatic hyperplasia in mice induced by subcutaneous injection of testosterone propionate is used as an experimental model of prostatic hyperplasia, prostatitis in rats induced by intra-prostatic injection of carrageenan is used as a non-bacterial prostatitis model, and prostatic hyperplasia induced by intra-prostatic injection of Staphylococcus aureus is used. Rat prostatitis is used as a model of bacterial prostatitis, and the effect of pomegranate peel and pomegranate seed extract on the prostate weight, prostate index, serum testosterone (testosterone) level, and plasma antioxidant capacity index (oxygenradical absorbance capacity, ORAC ), malondialdehyde (malondialdehyde, MDA) levels in the prostate, and nitric oxide (NO) levels in plasma. Evaluation of pomegranate peel extract and pomegranate seed extract on prostatic lecithin body density and white blood cell count, activity of acid phosphatase (PACP) in plasma, malondialdehyde (MDA) in prostate, NO level, Zn Content and effect of the antioxidant capacity index ORAC. The results showed that the extracts of pomegranate peel and pomegranate seeds had significant improvement effects on benign prostatic hyperplasia in mice and prostatitis in rats.

According to the above experiments, it can be proved that the pomegranate peel and pomegranate seed extract provided by the present invention is a safe and natural medicine, food and food additive, through anti-oxidation, regulating NO in the body, etc., so as to achieve the purpose of anti-prostatic hyperplasia and anti-prostatitis .

In addition, the present invention also suggests that pomegranate peel and pomegranate seed extracts can also be used as antioxidants or antioxidant composition raw materials for relieving and improving inflammatory reactions caused by benign prostatic hyperplasia and prostatitis. Since the pomegranate peel and pomegranate seed extracts provided by the invention are natural sources, and folks have long-term consumption experience, they are suitable for people of all ages and health conditions. Therefore, pomegranate peel and pomegranate seed extracts can provide multiple uses as health food, food additives. Although the anti-inflammatory effects of pomegranate peel and pomegranate seed extracts are known, the anti-hyperplasia and prostatitis activity exhibited by pomegranate peel and pomegranate seed extracts is demonstrated for the first time in the present invention.

Detailed ways:

The present invention can be illustrated in detail by the following examples, but is not limited only to the following examples.

Embodiment one: the preparation of pomegranate peel extract

Take the freshly dried pomegranate peel, crush it completely, add 10 times the amount of 70% ethanol, heat and reflux at 80°C for 3 hours, extract 3 times in total, combine the filtrate after filtration, concentrate under reduced pressure at 45°C and freeze-dry, the recovery rate is about 42.34% pomegranate peel extract was used in the following examples of the present invention.

Embodiment two: the preparation of pomegranate seed extract

Take the dried pomegranate seed powder, extract with 5 times the amount of acetone under reflux at 60°C for 3 hours, extract 3 times in total, combine the filtrate after filtration, and concentrate under reduced pressure at 45°C to obtain the pomegranate seed extract with a recovery rate of about 14.54%. In the following examples of the present invention.

Example 3: Inhibitory effect of pomegranate peel and pomegranate seed extract on 5-alpha reductase in vitro

Testosterone propionate 30 μM, NADPH 30 μM, and enzyme extract 6 mg/ml were mixed thoroughly and then added to samples of different concentrations prepared in Examples 1 and 2, reacted at 37° for 10 minutes, and the detection condition was 340 nm wavelength. Select the 15 μg/ml finasteride group as the control group to evaluate the relative inhibitory effect of pomegranate peel and pomegranate seed extract on the enzyme activity, (see accompanying drawing 1, 2)

Example 4: Effects of Pomegranate Peel and Pomegranate Seed Extract on Prostate Wet Weight and Prostate Index (PI) in Mice with Prostatic Hyperplasia Induced by Testosterone

The mice were randomly divided into blank control group, testosterone-induced benign prostatic hyperplasia model control group, positive drug Proscar group (19.47mg/kg), low-dose pomegranate peel extract group (250mg/kg), high-dose pomegranate peel extract group (500mg/kg), pomegranate seed extract low dose group (0.12% of food intake), pomegranate seed extract high dose group (1.2% of food intake), 10 mice in each group. The mice in each group except the blank group were subcutaneously injected with testosterone propionate (7.5 mg/kg) dissolved in olive oil every day for 12 consecutive days, and administered by intragastric administration at the same time. The blank and model groups were given the same volume of aqueous solution, fasted 24 hours after the last administration, anesthetized with ether, stripped the prostate, weighed, and calculated the wet weight of the prostate and the prostate index (PI). Experimental results The experimental data are expressed as x±s, and the Student t-test test is used for statistical processing. The results showed that, compared with the normal control group mice, the prostate wet weight and PI of the testosterone-induced benign prostatic hyperplasia model mice were significantly increased (P<0.01). Compared with the benign prostatic hyperplasia model group, the Proscar group, the pomegranate peel extract high and low dose group, and the pomegranate seed extract high and low dose group had significantly decreased prostate wet weight and prostate index (P<0.05 or P<0.01), And there was a dose-effect relationship with drug concentration (Table 1).

Table 1 Effects of pomegranate peel and pomegranate seed extracts on the wet weight and prostate index of the prostate in mice with testosterone-induced benign prostatic hyperplasia

^## p<0.01, compared with the blank group; ^* p<0.05, ^** p<0.01, compared with the testosterone model control group, it is statistically significant.

Embodiment five: the influence of pomegranate peel and pomegranate seed extract on testosterone-induced prostatic hyperplasia mouse prostate tissue malondialdehyde (MDA) level

Experimental animals, experimental methods, experimental groups and dosages are the same as in Example 4. Prostate tissue samples were taken, and 10% prostate tissue homogenate was prepared with physiological saline, centrifuged at 10,000 rpm for 10 minutes, and then the supernatant of the tissue homogenate was taken to determine the malondialdehyde content therein. The principle of determination is that malondialdehyde can condense with thiobarbituric acid (TBA) to form a red product, which is measured at 532nm using a microplate reader (Raybo MK3 microplate reader from Finland). Compared with the normal control group mice, the malondialdehyde (MDA) level in the prostate tissue homogenate of the benign prostatic hyperplasia model mice was significantly increased (P<0.01). The high and low dose group of skin extract and the high and low dose group of pomegranate seed extract can significantly reduce the malondialdehyde (MDA) level in the mouse prostate tissue homogenate (P<0.05 or P<0.01), and the decrease of the pomegranate peel extract group There is a dose-effect relationship with drug concentration (Table 2).

Table 2 Effects of pomegranate peel and pomegranate seed extracts on MDA levels in the prostate of mice with testosterone-induced benign prostatic hyperplasia

^## p<0.01, compared with the blank group; ^* p<0.05, ^** p<0.01, compared with the testosterone model control group, it is statistically significant.

Example 6: Effects of Pomegranate Peel and Pomegranate Seed Extract on Serum Testosterone in Mice with Prostatic Hyperplasia Induced by Testosterone

Experimental animals, experimental methods, experimental groups and dosages are the same as in Example 4. Blood was collected from the heart of the mice under ether anesthesia, and the plasma was separated by centrifugation at 5000 rpm for 5 minutes in a heparin-treated centrifuge tube. The level of serum testosterone in mice was determined by radioimmunoassay. The experimental results proved that, compared with the normal control group mice, the serum testosterone level of the testosterone-induced benign prostatic hyperplasia mice was significantly increased (P<0.01). Compared with the testosterone-induced benign prostatic hyperplasia model group, the Proscar group, the high-low dose group of pomegranate peel extract, and the high-low dose group of pomegranate seed extract could significantly reduce the level of serum testosterone in mice (P<0.01), and the drug concentration There was a dose-effect relationship (Table 3).

Table 3 Effects of pomegranate peel and pomegranate seed extracts on serum testosterone levels in mice with testosterone-induced benign prostatic hyperplasia

^## p<0.01, compared with the blank group; ^** p<0.01, compared with the testosterone model control group, which is statistically significant.

Embodiment 7: The effect of pomegranate peel and pomegranate seed extract on testosterone-induced prostatic hyperplasia mice plasma antioxidant capacity index (ORAC)

Experimental animals, experimental methods, experimental groups and dosages are the same as in Example 4. Blood was collected from the heart of the mouse under ether anesthesia, and then placed in a heparin-treated centrifuge tube, centrifuged at 5000 rpm for 5 minutes, and then the plasma was separated. After removing protein with 3% perchloric acid, centrifuge at 12,000 rpm for 15 minutes to recover the supernatant for ORAC analysis. Plasma antioxidant capacity index (ORAC) was analyzed using a multifunctional fluorescence analyzer (Tecan Genios, Austria), the excitation and emission wavelengths were 485nm and 525nm, respectively, and the oxidant attacking fluorescein (disodium fluorescein) was using 2,2'-azobis-2 -amidinopropane-dihydrochloride (AAPH; Wako Pure Chemical Industries, Ltd. Osaka, Japan). The experimental results were statistically processed by the t test. Compared with the normal control group mice, the plasma antioxidant capacity index of the testosterone-induced benign prostatic hyperplasia mice was significantly lower (P<0.05), compared with the testosterone-induced benign prostatic hyperplasia model group mice, Proscar group, pomegranate peel extract high and low dose group, and pomegranate seed extract high and low dose group can increase the plasma antioxidant capacity index of mice, that is, the ability to resist reactive oxygen species is significantly enhanced (P<0.01), and has Certain dose-effect relationship (Table 4).

Table 4 Effect of pomegranate peel and pomegranate seed extract on plasma antioxidant capacity index (ORAC) of testosterone-induced benign prostatic hyperplasia mice

^# p<0.05, compared with the blank group; ^** p<0.01, compared with the testosterone model control group, which is statistically significant.

Example 8: Effects of Pomegranate Peel and Pomegranate Seed Extract on Nitric Oxide (NO) Levels in Plasma of Testosterone-Induced Prostatic Hyperplasia Mice

Experimental animals, experimental methods, experimental groups and dosages are the same as in Example 4. As a highly reactive free radical in organisms, nitric oxide (NO) has dual functions of oxidation and anti-oxidation, as well as the function of second messenger and neurotransmitter, and it is also an effector molecule that mediates Regulate various pathological and physiological processes such as inflammatory response, cytotoxic effect and immune injury. Adopt Griess method, utilize 1% p-aminobenzenesulfonic acid, 0.1% N-(1-naphthyl)-ethylenediamine, 5% phosphoric acid to carry out diazo, azo reaction, generate red product, measure colorimetrically at 550nm, Sodium nitrite is used as a standard, and finally converted into NO ³ -concentration. The experimental results proved that compared with the normal control group mice, the plasma NO level of the testosterone-induced benign prostatic hyperplasia mice was significantly increased (P<0.01). Compared with the testosterone-induced benign prostatic hyperplasia model The high and low dose groups of extract and the high and low dose group of pomegranate seed extract can significantly reduce the plasma NO level of mice (P<0.05), and there is a certain dose-effect relationship (Table 5).

Table 5 Effects of pomegranate peel and pomegranate seed extracts on plasma NO levels in mice with testosterone-induced benign prostatic hyperplasia

^# p<0.05, compared with the blank group; ^* p<0.05, compared with the testosterone model control group, it is statistically significant.

Example 9: Effects of Pomegranate Peel and Pomegranate Seed Extract on Lecithin Body Density and White Blood Cell Count in Prostatitis Rat Model

Rats were randomly divided into blank control group, prostatitis model control group, positive drug Qianliekang group (500mg/kg), pomegranate peel extract low dose group (250mg/kg), pomegranate peel extract high dose group (500mg/kg ), pomegranate seed extract group (0.12% of food intake), pomegranate seed extract group (1.2% of food intake), 8 rats in each group. For groups other than the blank group, 7 days after the operation, 0.2ml of 1% carrageenan prepared with distilled water was injected directly into the dorsal lobe of the prostate, and then the peritoneum and skin were sutured to prepare a non-bacterial prostatitis model. Continue the administration for 3 days, ether anesthetize 40 minutes after the last administration, carefully separate each lobe of the prostate, weigh the prostate with an electronic balance, add sterile saline at 4ul/mg, homogenate, and record the density of lecithin bodies under a microscope and white blood cell count. Bacterial prostatitis model preparation, take 0.2ml of 10 ⁷ /ml Staphylococcus aureus prepared with normal saline and inject directly into the dorsal lobe of the prostate to replicate the bacterial prostatitis model. The experimental methods and experimental groups are the same as those for the non-bacterial prostate inflammation model. The experimental data are expressed as x±s, and the experimental results are statistically processed by the Studentt-test test.

Lecithin bodies and white blood cells are the two main aspects of prostatic fluid examination. When inflammatory lesions occur in the prostate, macrophages engulf lecithin bodies in large quantities, and the number of white blood cells increases at the same time. Therefore, recording lecithin body density and white blood cell count has important reference value for the diagnosis of prostatitis. The experimental results proved that compared with the normal control group rats, the lecithin corpuscle density of prostatitis model rats decreased significantly (P<0.01), and the number of white blood cells increased significantly (P<0.01), which was larger than that of the prostatitis model group. Compared with Qianliekang group, pomegranate peel extract high and low dose group, and pomegranate seed extract high and low dose group, the density of prostatic lecithin corpuscles was significantly increased (P<0.01), and the number of white blood cells was significantly reduced (P<0.01) , and has a certain dose-effect relationship (Table 6).

Table 6 Effect of pomegranate peel and pomegranate seed extract on lecithin body density and white blood cell count in prostatitis rat model

^## p<0.01, compared with the blank group; ^* p<0.05, ^** p<0.01, compared with the prostatitis model control group, it has statistical significance

Example 10: Effect of Pomegranate Peel and Pomegranate Seed Extract on Serum Acid Phosphatase Activity of Prostatitis Rat Model

Experimental animals, experimental methods, experimental groups and dosages are the same as in Example 9. 40 minutes after the last administration, blood was collected from the heart of the rats under ether anesthesia and left to stand for 1 hour, and then the serum was separated by centrifugation at 3000 rpm for 15 minutes at 4°C and the prostatic acid phosphatase activity in the serum was measured. Serum prostatic acid phosphatase activity was measured using a prostatic acid phosphatase test kit. The principle of determination is that prostatic acid phosphatase catalyzes the hydrolysis of α-naphthol phosphate to produce α-naphthol and inorganic phosphate, and α-naphthol reacts with diazonium salts to generate colored azo compounds, which are colorimetrically measured and calculated at 530nm. Acid phosphatase activity. The experimental results take 1 L of serum reacting with the substrate at 37°C to produce 1 μmol of free phenol per minute as an activity unit. The experimental results prove that, compared with the normal control rats, the acid phosphatase of the prostatitis rats is significantly increased (P<0.01). Compared with the prostatitis rat model group, the activity of plasma acid phosphatase in the Qianliekang group, the high and low dose groups of pomegranate peel extract, and the high and low dose group of pomegranate seed extract all decreased to varying degrees (P<0.05), and had Certain dose-effect relationship (Table 7).

Table 7 Effect of pomegranate peel and pomegranate seed extract on serum acid phosphatase activity in prostatitis rat model

^## p<0.01, compared with the blank group; ^* p<0.05, compared with the prostatitis model control group, it has statistical significance

Example 11: Effects of Pomegranate Peel and Pomegranate Seed Extracts on Prostate Tissue MDA Levels in Prostatitis Rat Models

Experimental animals, experimental methods, experimental groups and dosages are the same as in Example 9. The experimental results prove that, compared with the normal control rats, the MDA level in the prostatic tissue of rats with prostatitis is significantly increased (P<0.05). Compared with the rats in the prostatitis model group, the MDA levels in the prostate tissue of the rats in the Qianliekang group, the high-low dose group of pomegranate peel extract, and the high-low dose group of pomegranate seed extract were significantly reduced (P<0.05 or P<0.01), and There is a certain dose-effect relationship (Table 8).

Table 8 Effects of pomegranate peel and pomegranate seed extract on MDA level in prostatitis rat model prostate tissue

^# p<0.05, compared with the blank group; ^* p<0.05, ^** p<0.01, compared with the prostatitis model control group, there is statistical significance

Example 12: Effects of Pomegranate Peel and Pomegranate Seed Extracts on NO Levels in Prostate Tissue of Rats with Prostatitis

Experimental animals, experimental methods, experimental groups and dosages are the same as in Example 9. The experimental results proved that compared with the normal control group rats, the NO level in the prostate tissue of the non-bacterial prostatitis model rats was significantly increased (P<0.01). Compared with the non-bacterial prostatitis model rats, Qianliekang The levels of NO in the prostate tissue of rats in the control group, the high and low dose group of pomegranate peel extract, and the high and low dose group of pomegranate seed extract were significantly reduced (P<0.05 or P<0.01). In addition, compared with the rats in the normal control group, the NO level in the prostate tissue of the bacterial prostatitis model rats was significantly lower (P<0.05). The levels of NO in the prostate tissue of rats in the high and low dose groups of pomegranate seed extract and the high and low dose group of pomegranate seed extract were significantly increased (P<0.05 or P<0.01), and there was a certain dose-effect relationship (Table 9).

Table 9 Effect of pomegranate peel and pomegranate seed extract on NO level in prostatitis rat prostate tissue

^## p<0.01, compared with the blank group; ^* p<0.05, ^** p<0.01, compared with the prostatitis model control group, it has statistical significance

Example 13: Effects of Pomegranate Peel and Pomegranate Seed Extract on Zinc (Zn) Content in the Prostate Tissue of Prostatitis Rats

Experimental animals, experimental methods, experimental groups and dosages are the same as in Example 9. The experimental method is to cut the prostate tissue into pieces with scissors, add physiological saline at every 100 mg/mL, place it in an ice bath, and use a tissue homogenizer to make a 10% homogenate. After the homogenate was centrifuged at 3000rpm for 10min at 4°C, the supernatant of the homogenate was diluted 50 times with distilled water, detected by TAS-990 atomic absorption spectrophotometer, and the Zn content in the prostate tissue homogenate was calculated by the standard curve method . The experimental results proved that, compared with the normal control rats, the Zn content in the prostate tissue of the prostatitis rats was significantly reduced (P<0.01). Dose group, pomegranate seed extract high and low dose group, and rat prostate tissue Zn content were all significantly increased (P<0.05 or P<0.01), and there was a certain dose-effect relationship (Table 10).

Table 10 Effects of pomegranate peel and pomegranate seed extracts on zinc (Zn) content in prostatic tissue of rats with prostatitis.

^## p<0.01, compared with the blank group; ^* p<0.05, compared with the prostatitis model control group, it has statistical significance

Example 14: Effects of Pomegranate Peel and Pomegranate Seed Extract on Antioxidant Capacity Index (ORAC) in Prostatitis Rat Prostate Tissue

Experimental animals, experimental methods, experimental groups and dosages are the same as in Example 9. The experimental results prove that, compared with the normal control rats, the antioxidant capacity index in the prostate tissue of the prostatitis rats is significantly lower (P<0.01). Compared with the rats in the prostatitis model group, the antioxidant capacity index of the prostate tissue of the rats in the Qianliekang group, the high and low dose groups of pomegranate peel extract, and the high and low dose group of pomegranate seed extract were all significantly increased (P<0.05 or P<0.01 ), and has a certain dose-effect relationship (Table 11).

Table 11 Effect of pomegranate peel and pomegranate seed extract on prostatic tissue antioxidant capacity index (ORAC) of prostatitis rats

^## p<0.01, compared with the blank group; ^* p<0.05, ^** p<0.01, compared with the prostatitis model control group, it has statistical significance

Embodiment 15: the activity of the in vitro antioxidant capacity index (ORAC) of pomegranate peel and pomegranate seed extract

In the in vitro anti-oxidation experiment, different concentrations of pomegranate peel and pomegranate seed extracts were directly added to a 96-well microtiter plate to observe the delayed effect of the sample inhibiting the oxidant AAPH on the fluorescence decay of disodium fluorescein. As shown in Figure 3, both pomegranate peel and pomegranate seed extracts can delay the quenching speed of fluorescent substances by active oxygen free radicals in vitro, and have anti-active oxygen free radical effects. The anti-reactive oxygen free radical effect of pomegranate peel and pomegranate seed extract may come from small molecular polyphenolic compounds and punicic acid in pomegranate seed extract.

The invention uses subcutaneous injection of testosterone propionate to induce prostatic hyperplasia in mice as an experimental model of benign prostatic hyperplasia, intra-prostatic injection of carrageenan-induced rat prostatitis as a non-bacterial prostatitis model, and intra-prostatic injection of Staphylococcus aureus-induced rat prostate Bacterial prostatitis model, evaluation of pomegranate peel and pomegranate seed extract anti-prostatic hyperplasia and anti-prostatitis effect. The results showed that different doses of pomegranate peel and pomegranate seed extracts could effectively reduce the MDA level in prostate tissue homogenate, improve the body's antioxidant capacity, and regulate the NO level. Therefore, it can be speculated that pomegranate peel and pomegranate seed extracts can alleviate and improve the effects of prostate hyperplasia and prostatitis-induced prostate damage, which may be achieved through anti-oxidation, preventing peroxidative damage, and reducing the generation of lipid peroxides. . In addition, the present invention also suggests that pomegranate peel and pomegranate seed extracts can be used as antioxidants or antioxidant composition raw materials for the prevention and auxiliary treatment of some diseases caused by oxidative stress. Based on the above results, it was confirmed that the new application of the pomegranate peel and pomegranate seed extract of the present invention was established.

According to the above experiments, it can be proved that the extracts of pomegranate peel and pomegranate seeds provided by the present invention are safe and natural through anti-oxidation, so as to achieve anti-inflammatory effect, relieve and prevent, improve and treat prostatic hyperplasia and prostatitis, and can be used for the preparation of long-term Drugs, food additives, health foods and their components that are safe and non-toxic and have no side effects. Since the pomegranate peel and pomegranate seed extracts provided by the present invention are natural sources, and the folks have long-term eating experience, they are not only suitable for ordinary adults, but also for the elderly, children and those with low physical strength. Therefore, the pomegranate peel and pomegranate seed extracts can be used as health food and food additives to provide multiple purposes. According to the above experiments, it can be proved that the pomegranate peel and pomegranate seed extract provided by the present invention is a highly safe medicine, food and food additive.

Description of drawings

The inhibitory effect of Fig. 1 pomegranate peel extract on 5-alpha reductase in vitro: the abscissa is the pomegranate peel extract concentration (mg/ml), the experimental data is represented by x ± s, and the ordinate is the relative inhibition rate (%).

Fig. 2 Inhibitory effect of pomegranate seed extract on 5-alpha reductase in vitro: the abscissa is the concentration of pomegranate seed extract (mg/ml), the experimental data is represented by x ± s, and the ordinate is the relative inhibition rate (%).

The activity of the in vitro antioxidant capacity index (ORAC) of Fig. 3 pomegranate peel and pomegranate seed extract: the abscissa is time (minute), and the ordinate is relative fluorescence intensity. The principle is that sodium fluorescein emits 527nm fluorescence under the excitation of 485nm light, which can be oxidized by peroxyl radicals released by AAPH to make the fluorescence characteristics disappear. When antioxidants exist, they can compete with sodium fluorescein for oxidants and slow down the rate of fluorescence fading. The control (-AAPH) represents the fluorescence decay curve when the free radical generator AAPH is not added; the control (AAPH) represents the fluorescence decay curve when the free radical generator AAPH is added but no sample is added; the other two curves are: adding free radicals After generating the agent AAPH, the fluorescence decay curves when 0.25mg/ml pomegranate peel extract and 0.25mg/ml pomegranate seed extract were added to the samples respectively.

Claims (7)

1. The use of pomegranate peel and pomegranate seed extracts as active ingredients for the preparation of drugs for anti-prostatic hyperplasia, anti-prostatitis, and relief and improvement of their clinical symptoms. 2. Using pomegranate peel and pomegranate seed extracts as active ingredients to prepare health food and food additives for anti-prostatic hyperplasia and anti-prostatitis, and to alleviate and improve their clinical symptoms. 3. The use according to claim 1 or 2, characterized in that: the pomegranate peel extract contains polysaccharides and polyphenols active ingredients and pomegranate seed oil contains unsaturated fatty acid active ingredients, each gram of pomegranate peel The content of the polysaccharide and polyphenol active ingredients in the extract and the unsaturated fatty acid active ingredients in the pomegranate seed oil should not be less than 5 mg. 4. The use according to claim 1 or 2, characterized in that: said pomegranate peel and pomegranate seed extracts are made of water or any mixture of one or more of water, lower alcohols and polar solvents If the solvent is used for extraction, an appropriate amount of acid or alkali can also be added to the solvent for extraction. 5. The use according to claim 1, characterized in that: the medicine comprises an effective therapeutic dose of pomegranate peel and pomegranate seed extract, and a pharmaceutically acceptable carrier. 6. The use according to claim 1 or 2, characterized in that the use of the pomegranate peel and pomegranate seed extracts in various preparations of medicines, health foods or food additives can account for 0.001-100%. weight ratio. 7. The use according to claim 4, characterized in that: when extracting the pomegranate peel and pomegranate seed extract, sodium bicarbonate and/or sodium ascorbate may be added to the solvent.

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