1、 Pharmacopoeia standard of Polyporus umbellatus
This product is the dried sclerotia of Polyporus umbellatus (pers.) fries, a Polyporus family fungus. Excavate in spring and autumn to remove sediment and dry.
[properties] this product is strip-shaped, quasi round or flat block, some of which have branches, 5-25cm long and 2-6cm in diameter. The surface is black, gray black or brown black, shrunken or with tumor like protrusions. It is light and hard, with white or yellowish white cross-section and slightly granular shape. The air is weak and the taste is light.
[identification] (1) cut surface of this product: all are closely interwoven by hyphae. The outer layer is 27 ～ 54 μ m thick, and the hyphae are brown and difficult to separate; The internal hyphae are colorless, curved, 2-10 μ m in diameter, and some can be seen with transverse septa, branches or nodules. There are many calcium oxalate crystals between the hyphae, most of which are square octahedral, regular biconical octahedral or irregular polyhedral, with a diameter of 3-60 μ m and a length of 68 μ m, and sometimes several crystal sets.
(2) Take 1g of this product powder, add 20ml of methanol, sonicate for 30 minutes, filter, and take the filtrate as the test solution. Take ergosterol reference substance and add methanol to make a solution containing 1mg per 1ml as the reference solution. According to the test of thin-layer chromatography (general rule 0502), suck 20 μ l of the test solution and 4 μ l of the control solution, respectively dot on the same silica gel G thin-layer plate, use petroleum ether (60 ～ 90 ℃) - ethyl acetate (3:1) as the developing agent, develop, take out, dry, spray with 2% vanillin sulfuric acid solution, heat at 105 ℃ until the spots are clear. In the chromatogram of the test sample, spots with the same color appear at the corresponding position of the chromatogram of the control sample.
[inspection] the moisture content shall not exceed 14.0% (the second method of general rule 0832).
The total ash content shall not exceed 12.0% (general rule 2302).
Acid insoluble ash content shall not exceed 5.0% (general rule 2302).
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Chromatographic conditions and system suitability test octadecylsilane bonded silica gel was used as filler; Methanol was used as mobile phase; The detection wavelength was 283nm. The number of theoretical plates should not be less than 5000 according to the ergosterol peak.
Preparation of reference solution take an appropriate amount of ergosterol reference, weigh accurately, and add methanol to make a solution containing 50 μ g per 1ml.
Preparation of test solution take about 0.5g of powder (passing through No. 4 screen), weigh accurately, place in a corked conical flask, add 10ml of methanol precisely, weigh, sonicate (power 220W, frequency 50KHz) for 1 hour, cool, weigh again, make up the lost weight with methanol, shake well, filter, and take the filtrate.
The determination method is to precisely suck 20 μ l of the reference solution and 20 μ l of the test solution, inject them into the liquid chromatograph, and determine.
The content of ergosterol (c28h44o) in this product shall not be less than 0.070% calculated as dry product.
Decoction pieces
[processing] remove impurities, soak, wash, moisten, cut into thick pieces, and dry.
This product is round or irregular thick film. The outer skin is black or brownish black and wrinkled. The cut surface is white or yellowish white, slightly granular. The air is weak and the taste is light.
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The total ash content is the same as that of medicinal materials, and shall not exceed 10.0%.
[content determination] the content of ergosterol (c28h44o) in the same medicinal material shall not be less than 0.050%.
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[nature, taste and meridian tropism] sweet, light and flat. Return to kidney and bladder meridian.
[functions and indications] diuresis and dampness. It is used for urination, edema, diarrhea, drenching and purgation.
[usage and dosage] 6 ～ 12g.
[storage] store in a ventilated and dry place.

2、 Chemical constituents of Polyporus umbellatus

Sclerotia contains Polyporus umbellatus glucan Ⅰ, steroids: porostanol aPorosteroneB、PorosteroneC、PorosteroneD、PorosteroneE、PorosteroneF、PorosteroneG， 4,6,8 (14), 22-ergostatene-3-one, 25-deoxy-24 (28) - dehydroandrostone, 7,22-ergostadien-3-one, 7,22-ergostadien-3-ol, 5,7,22-ergostrien-3-ol, 5 α, 8 α - epidioxy-6,22-ergostadien-3-ol, and α - hydroxy tetracarbonate.
It was also reported that the polysaccharide in the fermentation filtrate of Polyporus umbellatus mycelium was composed of D-mannose, D-galactose and D-glucose, and the molar ratio was 20:4:1.

3、 Pharmacological effects of Polyporus umbellatus

1. diuretic use: Polyporus umbellatus decoction, equivalent to 0.25-0.5g / kg of crude drug, has obvious diuretic effect on non anesthetized dogs and can promote the excretion of sodium, chlorine, potassium and other electrolytes, which may be the result of inhibiting renal tubular reabsorption function.

2. impact on immune function:
2.1. immune enhancing effect: Polyporus umbellatus polysaccharide can significantly enhance the proliferative response of mouse T cells to ConA and the proliferative response of B cells to LPS. Polyporus umbellatus polysaccharide has obvious mitogenic effect on mouse whole spleen cells. At the dose of 12.5mg / (kg · day), Polyporus polysaccharide could significantly enhance the number of specific antibody secreting cells against SRBC in mice; . CTL is an important effector cell of immune surveillance, and plays a key role in tumor immunity.

2.2. a non-T cell mitogen action.
Splenocytes and thymocytes from normal mice, splenocytes from nude mice and splenocytes cultured after removing adsorptive cells were used to observe the proliferation response to different concentrations of Polyporus umbellatus polysaccharide. The results showed that the response of spleen cells of nude mice to Polyporus umbellatus polysaccharide was indistinguishable from that of normal mice cells, that is, T cells did not play a role in this reaction; After the adsorptive cells were removed from the splenocytes of nude mice, most of the remaining cells were B cells, but they could still proliferate significantly with Polyporus umbellatus polysaccharide, indicating that Polyporus umbellatus polysaccharide has the mitogenic effect of B cells. The response of splenocytes of nude mice to Polyporus umbellatus polysaccharide was significantly weakened after the removal of adsorptive cells, indicating that adsorptive cells have a certain auxiliary role in the response of splenocytes of nude mice to Polyporus umbellatus polysaccharide. . But it has no hemagglutination effect, which is different from mitogens from plants such as PHA and ConA.

2.3. effect on Mice 'blood ANAE positive lymphocytes: test mice were intraperitoneally injected with Polyporus umbellatus polysaccharide 2mg / 0.2ml every day for 7 consecutive days. Blood smears were taken before and after the experiment, and ANAE positive T lymphocytes were labeled by a-Naphthyl acetate esterase method, and the percentage was determined. The results showed that Polyporus umbellatus polysaccharide had no effect on the total number of ANAE positive T lymphocytes in the blood of mice, decreased the granular positive T lymphocytes, and significantly proliferated the dispersed granular positive T lymphocytes.

3. anti tumor effect:
3.1. effects on hepatic glucose metabolism and adrenal cortex function in H22 mice bearing liver cancer: Polyporus umbellatus polysaccharide was injected intraperitoneally into H22 mice bearing liver cancer at 200mg / kg × 2 days, and the animals were sacrificed on the 10th day. Ascites was removed, and the total number of tumor cells in each mouse was detected by eosin method. Results the inhibition rate was 39%. Intraperitoneal injection of 100-200mg / kg for 5 days increased the liver glycogen accumulation and the activities of gluconeogenic enzymes: glucose-6-phosphatase, fructose-1,6-bisphosphatase in cancer bearing mice, but had no effect on the liver of normal mice. 400mg / kg can restore the hyperfunctional cortical function of cancer bearing mice to normal. It is suggested that Polyporus umbellatus polysaccharide can adapt to the original, which may be a pharmacological basis for its anti-tumor effect.

3.2. effects on hepatic glycogen accumulation, gluconeogenesis and catabolic enzyme system in H22 mice bearing liver cancer: the regulation of gene expression and gene rearrangement during tumor formation are carried out through key enzymes, because gluconeogenesis is an important mechanism for the body to maintain homeostasis. The activities and dynamic changes of several key enzymes in the carbohydrate metabolism pathway were selected as indicators to observe how Polyporus umbellatus polysaccharide, which is effective for tumor, increases the accumulation of liver glycogen in cancer bearing mice. . However, it has no effect on the activity of glycogenolytic enzyme, namely phosphorylase. It is suggested that the liver glycogen accumulation of cancer bearing mice after Polyporus umbellatus polysaccharide administration plays a role by increasing the activity of gluconeogenic enzyme, accelerating gluconeogenesis and improving the self stable state of the body.

3.3. antitumor effect of Polyporus umbellatus extract: Polyporus umbellatus extract (mainly Polyporus umbellatus polysaccharide) has a significant inhibitory effect on transplanted tumor S-180 in mice. . About 6-7% of the tumor bearing mice treated with the extract completely regressed. For mice whose tumors completely subsided, the tumor cells were inoculated after 1-6 months, and none of them grew tumors. The extract could inhibit the growth of tumors at a certain dose by intraperitoneal injection, intravenous injection and gavage, but the effect of gavage was worse than that of intraperitoneal injection and intravenous injection, and the dosage was also large. Prophylactic administration has tumor suppressive effect on S-180. In the dose where chemotherapy alone does not show anti-tumor effect, adding an appropriate amount of Polyporus umbellatus extract will have significant anti-tumor effect. . This product can increase the cAMP content in S-180 ascites cancer cells. In the experimental group with high inhibition rate of cancer cell proliferation, the increase rate of cAMP content in cancer cells is also high. Generally, the cAMP content in cancer cells is lower than that in normal cells. The worse the canceration, the lower the camp content. Camp can transform tumor cells into normal cells. Grifolone A-G, seven compounds have cytotoxic effects on L1210 cells, and have dose-dependent inhibitory effects.

3.4. inhibitory effect on experimental bladder tumor: female rats were gavaged with 0.25ml (90mg) of carcinogen BBN [N-butyl-N - (4-hydroxybutyl) nitrosamine] solution, twice a week for 12 weeks, with a total dose of 2.16G BBN per rat, and were fed with 90g / kg Polyporus umbellatus dry powder at the same time. They were sacrificed after 30 weeks. .

4. protective effect on liver of mice with toxic hepatitis: carbon tetrachloride and D-galactosamine were intraperitoneally injected into mice to induce toxic hepatitis. Polyporus umbellatus polysaccharide 100-200mg / kg was intraperitoneally injected before and after induction, and the drugs were administered every 4, 8, and 12 hours. The activity of SGPT decreased, and the activities of liver 5 '- nucleotidase and acid phosphoaminase 6-phosphate glucose phosphatase increased. It also has a similar effect in vitro, indicating that it has a significant protective effect on the liver.

5. anti radiation effect: Polyporus umbellatus polysaccharide has obvious effect of preventing and treating acute radiation disease in mice, and the effective dose and time are relatively wide. Intraperitoneal injection of mice receiving a lethal dose (800rad) of total body irradiation 2 and 48 hours before irradiation could increase the survival rate by 30-70%. Administration after irradiation, whether oral or intraperitoneal, has protective potency, and prevention is more effective than treatment. . It is preliminarily believed that the anti radiation effect of Polyporus umbellatus polysaccharide may be through regulating the function of pituitary adrenal system, making the body in a state of stress, thereby enhancing the ability to resist radiation injury.

6. other effects: Polyporus umbellatus polysaccharide injected intraperitoneally into mice for 48 hours can significantly enhance the incorporation of 3H TDR in mouse thymocytes and accelerate the release of thymocytes. On the one hand, these phenomena no longer exist after adrenalectomy, and on the other hand, polysaccharides can significantly increase the content of corticosterone in animal plasma, so it can be considered that these effects are achieved through the adrenal cortex.