CN106063800B - Application of antrodia camphorata in anti-prostatic hyperplasia drugs - Google Patents
Application of antrodia camphorata in anti-prostatic hyperplasia drugs Download PDFInfo
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- CN106063800B CN106063800B CN201610620474.5A CN201610620474A CN106063800B CN 106063800 B CN106063800 B CN 106063800B CN 201610620474 A CN201610620474 A CN 201610620474A CN 106063800 B CN106063800 B CN 106063800B
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- antrodia camphorata
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/06—Fungi, e.g. yeasts
- A61K36/07—Basidiomycota, e.g. Cryptococcus
Abstract
The application of the antrodia camphorata in the preparation of the medicament for inhibiting the prostatic hyperplasia can reduce the height of prostatic epithelial cells, reduce the contents of serum ACP and DHT and reduce the expression of vascular endothelial growth factors in the prostate. The preparation method is simple, convenient and quick. The antrodia camphorata extract has the effect of treating prostatic hyperplasia, and the effect can be comparable to that of the western medicine finasteride; meanwhile, the traditional Chinese medicine composition has obvious anti-inflammatory and analgesic effects, can inhibit the mouse cotton ball granulation swelling and reduce the acetic acid writhing frequency of the mouse, and can be used for relieving inflammation and pain accompanied by prostatic hyperplasia.
Description
Technical Field
The invention relates to a technology in the field of medicine preparation, in particular to an application of antrodia camphorata in an anti-prostatic hyperplasia medicine.
Background
Prostatic Hyperplasia (BPH) is a common disease in middle-aged and elderly men, and the incidence rate of BPH increases year by year with the aging of the population. Statistics show that the incidence rate of men aged 51 to 60 years in China is up to more than 70%. The main clinical manifestations of prostatic hyperplasia are a series of lower urinary tract symptoms, such as frequent micturition, thin urinary line, laboursome urination, urine retention, etc., and severe patients can cause the symptoms of the lower urinary tract to worsen and the upper urinary tract to be damaged or hydronephrosis, renal failure (LUTS). With regard to the pathogenesis of prostatic hyperplasia, it is believed that prostatic hyperplasia is associated with endocrine dysregulation of body hormones, overexpression of growth factors, and overexpression of proto-oncogenes. In addition, prostatic hyperplasia is accompanied by inflammation and pain.
The treatment method of the prostatic hyperplasia comprises operation treatment and drug treatment, wherein the operation treatment has good effect, but causes physical trauma to patients, and the acceptance is not high, according to the action mechanism and the types of the drugs, the current commonly used drugs for treating the prostatic hyperplasia are generally divided into three types, namely 5 α -reductase inhibitors, receptor blockers, traditional Chinese medicines and plant preparations, western medicines have good effect but bring a plurality of adverse reactions, and the traditional Chinese medicine preparation and the plant extract have the defects of large administration dosage, long treatment course and slow response.
Antrodia cinnamomea (Antrodia cinnamomea T.T.Chang & W.N.Chou) is a Taiwan medicinal material, also named as Antrodia cinnamomea, Antrodia camphorata, Antrodia cinnamomea, and Antrodia sanguinea, and belongs to the genus Antrodia of Aphyllophorales of class Basidiomycota. At present, more than two hundred chemical components are separated from antrodia camphorata, and comprise primary metabolites of fungi such as alkanal, aromatic hydrocarbon, carboxylic acid, fatty acid ester, adenosine and the like, and essential oil, polysaccharide, triterpene, superoxide dismutase, lectin, trace elements, benzene derivatives, android quinal and the like. Research shows that the crude extract of Antrodia camphorata has cytotoxic effect on cancer cells such as liver cancer, breast cancer, leukemia, pancreatic cancer and the like. However, the report of treating prostatic hyperplasia by Antrodia camphorata is not available.
Disclosure of Invention
The invention provides an application of antrodia camphorata in an anti-prostatic hyperplasia drug, aiming at the defects that the preparation in the prior art is complicated, various toxic solvents such as dichloromethane, methanol, acetone and the like are mostly used, the preparation process needs to use MPLC and other precise instruments, repeated column chromatography is needed, the cost is high, the yield is low and the like, and the preparation method is simple, convenient and quick. The antrodia camphorata extract has the effect of treating prostatic hyperplasia, and the effect can be comparable to that of the western medicine finasteride; meanwhile, the traditional Chinese medicine composition has obvious anti-inflammatory and analgesic effects, can inhibit the mouse cotton ball granulation swelling and reduce the acetic acid writhing frequency of the mouse, and can be used for relieving inflammation and pain accompanied by prostatic hyperplasia.
The invention is realized by the following technical scheme:
the invention relates to an application of antrodia camphorata in preparing a medicament for inhibiting prostatic hyperplasia.
The medicine can reduce the height of prostate epithelial cells, improve the pathological change of prostate tissue, obviously reduce the contents of ACP and DHT in serum, and reduce the expression of vascular endothelial growth factors in prostate.
The invention also relates to an application of the antrodia camphorata in preparing the anti-inflammatory analgesic medicament.
The Antrodia camphorata is prepared from Antrodia camphorata (Antrodia cinnamama T.T.Chang & W.N.Chou) fruiting body, and dry extract is obtained after grinding and ethanol reflux extraction and reduced pressure concentration, or dry suspension or suspension is obtained through ethanol recrystallization after ethanol reflux extraction, concentration and drying.
Drawings
FIG. 1 is a graph showing the effect of Antrodia camphorata extract on prostate histopathology in BPH rats;
in the figure are HE staining at x 400: (a) blank, (b) is a model, (c) is finasteride, (d) is an antrodia camphorata low-dose group, (e) is an antrodia camphorata high-dose group;
FIG. 2 is a graph showing the effect of Antrodia camphorata extract on VEGF in prostate tissue of BPH rats;
in the figure, x 400 is: (a) blank, (b) model, (c) finasteride, (d) negative control, (e) low dose of antrodia camphorata, and (f) high dose of antrodia camphorata.
Detailed Description
Example 1
The present example prepares an antrodia camphorata extract for treating prostatic hyperplasia, which includes the following steps:
1) picking up fruiting bodies of Antrodia camphorata, removing impurities, and shearing or chopping;
2) putting the mixture into a round-bottom flask, and adding 65-90% ethanol serving as an extraction solvent, wherein the material-liquid ratio is 2-3 g crude drug/1000 mL;
3) reflux extraction is carried out for 30-60 min;
4) filtering the extractive solution, recovering the extraction solvent, and concentrating under reduced pressure to obtain dry extract.
The following experiments show that the antrodia camphorata extract prepared in the example has the effect of resisting the rat prostatic hyperplasia induced by testosterone propionate.
Experimental materials: test animals: male SD rats, 72, weight 160 ~ 180 g. Test drugs: finasteride suspension 0.25mg/mL (positive control group); antrodia camphorata extract suspension 16mg crude drug/mL (low dose Antrodia camphorata extract); antrodia camphorata extract suspension 48mg crude drug/mL (high dose Antrodia camphorata extract).
The test method comprises the following steps: preparing a rat prostatic hyperplasia model according to the guiding principle of the medicine effect research of the new medicine. Male SD rats were randomly divided into 5 groups of 8 rats each, which were: blank group, model group, finasteride group, and antrodia camphorata alcohol extract low and high dose group. One week of adaptive breeding before molding, one week later, the blank group was removed, and the rats in the other groups were injected with testosterone propionate 20mg/kg x d every other day for 3 consecutive weeks. After 3 weeks of molding, each administration group was subjected to intragastric administration, the normal group and the model group were subjected to intragastric administration with 5% CMC-Na, the administration volume was 0.4mL/100g, the finasteride group was subjected to intragastric administration with finasteride suspension (1 mg/kg), and each group of Antrodia camphorata was subjected to continuous intragastric administration for 30d once a day at a determined dose (72 mg/kg d in the low dose group and 216mg/kg d in the high dose group). During the administration period, the rats were observed for general behavioral activities, appetite, mental state, hair gloss, defecation, and the like. The rats in each group were anesthetized after the last administration for 2h, blood was taken from the abdominal aorta, the rats were sacrificed, the prostate was taken, and the wet weight of the prostate was weighed. Prostate index was calculated as follows: prostate index PI ═ (prostate weight/rat body weight) × 100.
Fixing the same parts of prostate ventral lobe with 4% paraformaldehyde, embedding in paraffin for sectioning, staining with hematoxylin-eosin, and inspecting pathological morphological change of rat prostate tissue section under light microscope. Prostate epithelial cell height was also measured using Image-pro plus 6.0.
Anesthetizing rat, collecting blood from abdominal aorta in vacuum blood collection tube, standing at room temperature for 1h, refrigerating at 4 deg.C overnight, centrifuging on day 2, and collecting supernatant and storing at-80 deg.C. After uniform thawing at room temperature, the contents of acid phosphatase (ACP) and Dihydrotestosterone (DHT) in rat serum are detected according to the kit steps.
Streptavidin-biotin complex (SABC) immunohistochemistry detects VEGF expression.
The experimental data are expressed as mean ± sd, and the data processing adopts SPSS 16.0 statistical software. Adopting One-way ANOVA as the measurement data; counting data is checked by adopting rank sum; p is less than 0.05, and has statistical significance.
Test results
1) Effect on prostate index
Table 1 shows the effect of antrodia camphorata extract on prostate index in rats with prostate hyperplasia. The Prostate Index (PI) of the model group rats is significantly higher than that of the blank group (p < 0.05), and the prostate index of the finasteride group rats is significantly lower than that of the model group (p < 0.05); the group of antrodia camphorata extracts tended to be smaller than the model group.
TABLE 1 Effect of Antrodia camphorata extract on prostate index in BPH rats (mean + -sd, n ═ 8)
#P < 0.05, compared to blank;*P＜0.05，**p < 0.01, compared to model group.
2) Influence on the content of acid phosphatase and dihydrotestosterone in serum of rat with prostatic hyperplasia
Table 2 shows the effect of Antrodia camphorata extract on the contents of acid phosphatase and dihydrotestosterone in the serum of rats with prostatic hyperplasia. Compared with the blank group, the model group has the advantages that the content of serum ACP and DHT is increased, and the difference is extremely obvious (p is less than 0.01); compared with the model group, the low-dose and high-dose groups of antrodia camphorata have low ACP and DHT contents in serum, and the difference is obvious (p is less than 0.05).
TABLE 2 Effect of Antrodia camphorata extract on ACP and DHT in serum of BPH rats (mean + -sd, n-8)
##P < 0.01, compared to blank;*P＜0.05，**p < 0.01, compared to model group.
3) Effect on prostate histomorphology in rats with prostate hyperplasia
FIG. 2 shows the effect of Antrodia camphorata extract on prostate histomorphology in rats with prostate hyperplasia. The normal group has clear arrangement of glands, clear and complete glandular cavity structure and prostate epithelial cells in single-layer columnar arrangement. The number of the glands in the model group is increased, the number density of the glands per unit area is increased, epithelial cells are obviously thickened, the structure of a part of the glands is multi-layer and is disorderly arranged, most of the glands are provided with papillary bulges, and interstitial fibrous tissues and smooth muscle tissues are increased, which indicates that the modeling is successful. The low-dose antrodia camphorata tissue cavities are obviously reduced, the luminal cells are mostly arranged in a single-layer column shape, and the interstitium is few and loose. The prostate gland epithelial cells of the antrodia camphorata high-dose group and the finasteride group are arranged in a single layer or a multiple layer mode, the epithelial cells are mostly in a high column shape or a cubic shape, and the papillary structures are obviously reduced (figure 1), which indicates that the antrodia camphorata high-dose group has the function of obviously inhibiting the hyperplasia of prostate.
The pathological morphology of the prostatic hyperplasia of each administration group of the experiment is measured according to a semi-quantitative standard, and the measurement result is shown in a table 3. By rank test, compared with the blank group, the model group has obvious pathological changes of prostatic hyperplasia (P is less than 0.01). Compared with a model group, the finasteride and antrodia camphorata high-dose group can obviously relieve the symptom of prostate hyperplasia of rats (P is less than 0.01) after being administrated; the low dose group of Antrodia camphorata also can relieve the symptom of prostate hyperplasia of rats to a certain extent (P is less than 0.05).
TABLE 3 Effect of Antrodia camphorata extract on BPH rat prostate tissue morphology (mean ± sd, n ═ 8)
"-" indicates that the prostate gland is essentially normal, the glandular epithelium is essentially normal, and the stroma is essentially normal; "+" indicates that the gland is essentially normal, the glandular epithelium is hyperplastic, and the stroma is essentially normal; "+ +" indicates glandular hyperplasia, glandular epithelial hyperplasia, interstitial hyperplasia; the "+++" indicates that the gland is obviously hyperplastic, the glandular epithelium is obviously hyperplastic, and the stroma is obviously hyperplastic.
4) Effect of Antrodia camphorata extract on prostate epithelial cell height of BPH rat
Table 4 demonstrates the effect of antrodia camphorata extract on prostate epithelial cell height in prostate-proliferating rats. The epithelial cell height of the model group was significantly increased (p < 0.05) compared to the blank group; compared with the model group, each group of antrodia camphorata can obviously reduce the height of prostate epithelial cells.
TABLE 4 Effect of Antrodia camphorata on prostate epithelial height in BPH rats (mean ± sd, n ═ 8)
##P < 0.01, compared to blank;*P＜0.05，**p < 0.01, compared to model group.
5) Effect of Antrodia camphorata extract on Vascular Endothelial Growth Factor (VEGF) expression of prostate tissue of BPH rats
VEGF expression of prostate tissues in a normal group is weakly positive; the expression of the model group is strong positive, the expression is that the number of positive cells is increased, most of the positive cells are dark brown yellow, and the staining is darker; the positive cells of the finasteride group are reduced and the staining becomes lighter compared with the positive cells of the model group; the situation of each administration group of the antrodia camphorata is improved compared with that of the model group, the staining is lighter, and the number of positive expression cells is reduced; see figure 2 for details.
6) Effect of Antrodia camphorata extract on VEGF expression in BPH rat prostate epithelial cells
Table 5 shows the effect of Antrodia camphorata extract on the mean optical density of VEGF in prostate epithelial cells of prostate-proliferating rats. The expression of VEGF is highest in the model group, the average optical density value is the largest, and the obvious difference is compared with the normal group (P is less than 0.01); the expression of VEGF after the Antrodia camphorata treatment is obviously reduced compared with the model group, and the average optical density value is reduced (P is less than 0.01).
TABLE 5 Effect of Antrodia camphorata extract on VEGF in BPH rat prostate (mean + -sd, n ═ 8)
#P < 0.05, compared to blank;*P＜0.05，**p < 0.01, compared to model group.
According to the test results, the antrodia camphorata extract can obviously reduce the prostate index of rats with prostatic hyperplasia induced by testosterone propionate, reduce the height of prostate epithelial cells, improve the pathological change of prostate tissues, obviously reduce the contents of ACP and DHT in serum and reduce the expression of vascular endothelial growth factors in prostate.
Example 2
In this example, an anti-inflammatory and analgesic antrodia camphorata extract was prepared in the same manner as in example 1, and was used to treat inflammation and pain associated with prostatic hyperplasia.
The following experiments show that the Antrodia camphorata extract prepared in the example has analgesic and anti-inflammatory effects.
Test animals: ICR mice, SPF grade, 18-22 g, male and female half.
Test method
1) Mouse writhing method
ICR mice were randomly divided into 8 groups: the normal saline group (NS), the positive control aspirin group (Asp40mg/kg), and the antrodia camphorata group (low dose group: 104mg/kg, medium dose group: 208mg/kg, high dose group: 416mg/kg) each contain 10 male and female halves. The mice of each group are respectively given physiological saline or corresponding drugs, and are subjected to intragastric administration for 1 time every day for 5 days continuously. After the last administration for 1h, each mouse was injected intraperitoneally with 0.2mL of 1% glacial acetic acid, resulting in a writhing response in the mice. The writhing reaction is shown as: the abdomen of the mouse is obviously concave, the hind limbs are extended backwards, the hip is raised and the like, and the whole torsion reaction is counted once. The frequency of writhing reaction is high within 15min after glacial acetic acid injection, the latency period is about 3min, therefore, the writhing times of mice within 15min after glacial acetic acid injection for 3min are taken as quantitative indexes of pain, the more the writhing times of the administration group is compared with that of the blank group, the more the analgesic effect is shown, the difference of the analgesic effect of the test drugs is reflected, and the inhibition rate of writhing reaction of the mice of each administration group is calculated.
2) Mouse cotton ball granuloma method
An ICR mouse is anesthetized with 0.1mL/10g of 10% chloric acid hydrate, a mouse plate is fixed, a 1cm long opening is cut in the back of the mouse under an aseptic condition, a 10mg sterilized cotton ball (which is subjected to autoclaving and penicillin sodium treatment in advance) is buried under the axilla of the right forelimb of the mouse and is sutured, and the penicillin sodium powder is coated outside the mouse to resist infection after operation. Mice were randomized into 8 groups (n ═ 10): blank group, positive control group (cortisone acetate 1.8 mg/kg. d), low, medium and high dose groups of Antrodia camphorata. The blank group was dosed at equal volume to the dexamethasone acetate group. Beginning the gavage administration the next day after the operation for 7d continuously; and after 1h of last administration, killing, stripping and taking out the granulation tissue of the cotton ball, placing in a constant-temperature drying box at 60 ℃ for 24h, weighing the weight of the cotton ball, subtracting the weight of the original cotton ball to obtain the dry weight of the proliferated granulation, taking the dry weight of the granulation as an evaluation index, and calculating the granulation swelling inhibition rate.
Test results
Tables 6 and 7 show that the three dose groups of the antrodia camphorata extract can obviously reduce the times of writhing reaction of mice caused by acetic acid and inhibit the proliferation of cotton ball granulation tissues of the mice compared with the normal saline group. The Antrodia Camphorata extract has antiinflammatory and analgesic effects, and can be used for treating inflammation and pain accompanied with prostatic hyperplasia.
TABLE 6 inhibitory effect of Antrodia camphorata extract on writhing response in acetic acid induced mice (mean + -sd, n ═ 10)
*P＜0.05，**P < 0.01, compared to the blank group.
TABLE 7 inhibition of mouse cotton ball granuloma by Antrodia camphorata extract (mean + -sd, n ═ 10)
*P＜0.05，**P < 0.01, compared to the blank group.
The foregoing embodiments may be modified in many different ways by those skilled in the art without departing from the spirit and scope of the invention, which is defined by the appended claims and all changes that come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein.
Claims (2)
1. The application of antrodia camphorata in preparing the medicine for inhibiting the hyperplasia of prostate is characterized in that the antrodia camphorata dry extract is used for preparing the medicine for inhibiting the hyperplasia of prostate;
the antrodia camphorata dry extract is prepared by the following steps:
1) picking up fruiting bodies of Antrodia camphorata, removing impurities, and shearing or chopping;
2) putting the mixture into a round-bottom flask, and adding 65-90% ethanol serving as an extraction solvent, wherein the material-liquid ratio is 2-3 g crude drug/1000 mL;
3) reflux extraction is carried out for 30-60 min;
4) filtering the extractive solution, recovering the extraction solvent, and concentrating under reduced pressure to obtain extract dry extract;
the inhibition of the prostatic hyperplasia refers to reduction of the height of human or rat prostatic epithelial cells, reduction of serum ACP and DHT contents and reduction of the expression of vascular endothelial growth factors in the prostate;
the Antrodia camphorata is prepared from Antrodia camphorata (Antrodia cinnamanomea T.T.Chang & W.N.Chou) fruiting body, and is ground, extracted by ethanol reflux and concentrated under reduced pressure to obtain dry extract.
2. The application of antrodia camphorata in preparing the medicine for resisting the hyperplasia of prostate is characterized in that the antrodia camphorata dry extract is used for preparing the medicine for resisting inflammation and easing pain;
the Antrodia camphorate is prepared by taking Antrodia camphorate (Antrodia cinnamanomea T.T.Chang & W.N.Chou) fruiting body, grinding, extracting by ethanol reflux, and concentrating under reduced pressure to obtain dry extract;
the Antrodia camphorata is prepared from Antrodia camphorata (Antrodia cinnamanomea T.T.Chang & W.N.Chou) fruiting body, and is ground, extracted by ethanol reflux, concentrated, dried and recrystallized by ethanol to obtain dry suspension or suspension.
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| JP2005110508A (en) * | 2003-10-02 | 2005-04-28 | Nec Soft Ltd | Growth factor-like protein derived from snake venom having specific binding property to vascular endothelial growth factor receptor 2 |
| CN101151957A (en) * | 2006-09-26 | 2008-04-02 | 国鼎生物科技股份有限公司 | Culture method and uses of antrodia cinnamomea |
| CN101219169A (en) * | 2007-01-08 | 2008-07-16 | 欧阳湘 | Externally used Chinese medicine for treating prostate gland hyperplasia and delaying human body caducity |
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