1、 Pharmacopoeia standard of Polygonum cuspidatum
This product is the dried rhizome and root of Polygonum cuspidatum sieb.et zucc. Excavate in spring and autumn, remove fibrous roots, wash, cut short sections or thick pieces while fresh, and dry them in the sun.
. There are septa or cavities in the rhizome pulp. It is hard. It is slightly bitter and astringent.
[identification] (1) the powder of this product is orange yellow. Calcium oxalate cluster crystals are extremely numerous and large, with a diameter of 30-100 µ M. the stone cells are yellowish, square like or round like, and some are branched. The branched stone cells are often 2-3 connected, with a diameter of 24-74 µ m, striated pores, and the cell cavity is filled with starch granules. Cork cells are polygonal or irregular in shape, and their cavities are filled with reddish brown substances. The diameter of the conduit with marginal pits is 56 ～ 150 µ M.
(2) Take 0.1g of this product powder, add 10ml of methanol, sonicate for 15 minutes, filter, evaporate the filtrate, add 5ml of 2.5mol/l sulfuric acid solution to the residue, heat it in a water bath for 30 minutes, cool it, shake it with chloroform twice, 5ml each time, combine the chloroform solution, evaporate to dryness, add 1ml of chloroform to dissolve the residue, and use it as the test solution. . Then take emodin reference substance and emodin methyl ether reference substance, add methanol to make a solution containing 1mg each in 1ml as the reference solution. Test according to thin-layer chromatography (general rule 0502), suck 4 µ l of the test solution and 1 µ l of the control drug solution, respectively, and dot them on the same silica gel G thin-layer plate. Use the upper solution of petroleum ether (30 ~ 60 ℃) - ethyl formate formic acid (15:5:1) as the developing agent, unfold, take out, dry, and view under the ultraviolet light (365nm). In the chromatogram of the test sample, fluorescent spots of the same color are displayed at the positions corresponding to the chromatogram of the control medicinal material and the chromatogram of the control sample; .
[inspection] the moisture content shall not exceed 12.0% (the second method of general rule 0832).
The total ash content shall not exceed 5.0% (general rule 2302).
Acid insoluble ash content shall not exceed 1.0% (general rule 2302).
[extract] according to the cold leaching method under the determination method of alcohol soluble extract (general rule 2201), the use of ethanol as solvent shall not be less than 9.0%.
[content determination] Rhein shall be determined by HPLC (general rule 0512).
Chromatographic conditions and system suitability test octadecylsilane bonded silica gel was used as filler; The mobile phase was methanol-0.1% phosphoric acid solution (80:20); The detection wavelength is 254nm. The number of theoretical plates should not be less than 3000 according to the emodin peak.
Preparation of reference solution take an appropriate amount of emodin reference substance dried under reduced pressure for 24 hours with phosphorus pentoxide as desiccant, weigh accurately, add methanol to make a solution containing 48 µ g per 1ml.
Preparation of test solution take about 0.1g of powder (passing through No. 3 screen), accurately weigh it, precisely add 25ml of chloroform and 20ml of 2.5mol/l sulfuric acid solution, weigh it, heat it in an 80 ℃ water bath and reflux it for 2 hours, cool it to room temperature, then weigh it, use chloroform to make up the lost weight, and shake it well. .
The determination method is to precisely suck 5 µ l each of the reference solution and the test solution, inject them into the liquid chromatograph, and determine.
The content of emodin (c15h10o5) in this product shall not be less than 0.60% according to the dry product.
Polydatin operates in the dark. .
Chromatographic conditions and system suitability test octadecylsilane bonded silica gel was used as filler; Acetonitrile water (23:77) was used as mobile phase; The detection wavelength was 306nm. The number of theoretical plates should not be less than 3000 according to the peak of polydatin.
Preparation of reference solution take an appropriate amount of polydatin reference substance dried under reduced pressure for 24 hours with phosphorus pentoxide as desiccant, weigh it accurately, and add dilute ethanol to make a solution containing 15 µ g per 1ml.
Preparation of the test solution take about 0.1g of the powder (passing through No. 3 screen), weigh accurately, add 25ml of dilute ethanol precisely, weigh, heat and reflux for 30 minutes, cool to room temperature, weigh again, make up the lost weight with dilute ethanol, shake well, take the supernatant, filter, and take the filtrate.
The determination method is to precisely suck 10 µ l of the reference solution and 10 µ l of the test solution, inject them into the liquid chromatograph, and determine.
The content of polydatin (c20h22o8) in this product shall not be less than 0.15% according to the dry product.
Decoction pieces
[processing] remove impurities, wash, moisten, cut into thick pieces, and dry.
[nature, taste and meridian tropism] slightly bitter and cold. It belongs to the liver, gallbladder and lung meridians.
[functions and indications] it can remove dampness and jaundice, clear away heat and toxin, relieve blood stasis and pain, relieve cough and phlegm. It is used for damp heat jaundice, drenching turbidity, band down, rheumatism, arthralgia, carbuncle, swelling and sore poison, water and fire scald, amenorrhea, symptomatic scrofula, traumatic injury, lung heat cough.
[usage and dosage] 9 ～ 15g. Apply an appropriate amount externally and make it into decoction or ointment.
[note] pregnant women should use it with caution.
[storage] put it in a dry place to prevent mold and moth.

2、 Chemical constituents of Polygonum cuspidatum

The roots and rhizomes contain free anthraquinone and anthranilin, mainlyEmodin（emodin），Emodin methyl ether（Physcion），Chrysophanol（chrysopha－nol）， Anthraglycoside A is 8-o - β - D-glucoside, anthraglycoside B is emodin-8-o - β - D-glucoside, charming alcohol, 6-hydroxyaloe emodin, emodin-8-o - β - D-glucoside, 6-hydroxyaloe emodin, emodin-8-methyl ether, 6-hydroxyaloe emodin-8-methyl ether, etc. It also contains stilbene compounds:Resveratrol(resveratrol) is 3,4 ', 5-trihydroxystilbene,PolydatinPolydatin, namely resveratrol-3-o - β - D-glucoside, also containsProtocatechuic acid（Protocate－chuic acid），Catechin（catechin)， 2,5-dimethyl-7-hydroxychromone, 7-hydroxy-4-methoxy-5-methyl coumarin, 2-methoxy-6-acetyl-7-methyljuglone, torachrysone-8-o-d-glucoside, β - sitosterol glucoside- sitosterol glucoside), glucose, rhamnose, polysaccharide, amino acid 12.99%, copper, iron, manganese, zinc Potassium and potassium salt, etc.

3、 Pharmacological action of Polygonum cuspidatum

1. effects on cardiovascular system anthraquinone injection has obvious hypotensive effect on anesthetized rabbits. After several consecutive administrations, the drug volume accumulates, and a small dose can cause sudden drop in blood pressure or even death. Anthraquinone injection could significantly slow down the heart rate of anesthetized rabbits, and no other changes in ECG were found. Resveratrol glucoside (PD) injection can double the reduced cardiac output and stroke index of hemorrhagic shock rats during shock, reduce the total peripheral resistance to near normal, restore the decreased cardiac output after burn to 91% of pre injury, achieve ventricular function of 100.1%, restore the systemic peripheral resistance to near normal, and significantly improve the survival rate of animals. PD has obvious positive inotropic effect on the isolated working heart of normal rats, but does not accelerate the heart rate. It can resist the negative inotropic effect of propranolol and heart failure caused by phenobarbital sodium. Pd0.05.0.15.0.45mmol / l could significantly accelerate the cell beating rate, and could be blocked by nisoldipine, propranolol and phentolamine. Pd0.05 and 0.15mmol/l could significantly reduce LDH release from cardiomyocytes at 6 and 9 hours after glucose deprivation and hypoxia injury, and pd0.15mmol/l could significantly reduce LDH release from cardiomyocytes at 9 hours after chlorpromazine injury. It is suggested that PD has a protective effect on cardiomyocytes injured by glucose deprivation, hypoxia and chlorpromazine. . In vivo experiments showed that pd5mg / kg significantly inhibited platelet aggregation induced by ADP and AA; It also has a certain inhibitory effect on Ca2 + - induced platelet aggregation, especially at 60 minutes after administration. . Pd10.4 μ g / ml and yohimbine 0.22 μ g / ml significantly inhibited the platelet aggregation induced by CN at a final concentration of 0.09-1.50 μ g / ml. PD can improve microcirculation and excite myocytes, and inhibit ADP induced platelet aggregation in rabbits in vitro and in vivo. PD can significantly inhibit human platelet aggregation induced by ADP and epinephrine, with the inhibition rate of 15.2-29.7% for the former and 21.9-45.9% for the latter. Pd6.7-107.2umol / L significantly inhibited AA and ADP induced platelet aggregation and TXB2 production in rabbits, and the inhibition rates of platelet aggregation were 48-90% and 43-69%, respectively; The inhibition rates of TXB2 production were 50-87% and 43-68%, respectively; There was a significant positive correlation between the inhibition rate of platelet aggregation and the inhibition rate of TXB2 production. . Pd1.71mmol / l could noncompetitively inhibit the effect of norepinephrine on pulmonary artery contraction by using rabbit ex vivo vascular volume method. 4.09mmol / L and 5.12mmol / l could dilate the isolated pulmonary artery of rabbits, and 5.12mmol / l could dilate the isolated carotid artery of rabbits. The relaxant effect of PD on pulmonary artery can be attenuated by propranolol, a b-adrenoceptor blocker. PD can change the contractile blood vessels into dilated ones after burn and reduce thrombosis. Effect of PD on blood pressure during irreversible hemorrhagic shock in rats: after intravenous injection of different doses of PD and reinfusion of the released blood, the arterial blood pressure can rise steadily, the narrowed arteriolar caliber during shock can be restored, the capillaries can be opened, and the animal survival rate can be significantly improved. The effect of PD has a certain trend with increasing dose.
2. hepatoprotective effect: PD and resveratrol have therapeutic effects on liver damage induced by oral feeding of peroxide corn oil in rats
.
2.2. reduce the levels of got and GPT, LPO, FFA, TC, TG and HDL Ch in the serum of mice; (3) Prevent peroxides from being induced by microsomal ADP and n-adph in rat liver. The experimental results of mice drinking corn oil cholesterol cholic acid acidic mixture showed that, PD、 Resveratrol affects lipid metabolism as:.
2.3. taking resveratrol has a certain inhibitory effect on the accumulation of TC and TG in the liver; PD has a certain inhibitory effect on the increase of serum TG and LDL ch; Reduced atherogenic index.
2.4. resveratrol and PD reduced lipogenesis of 14C palmitic acid in mouse liver. Polygonum cuspidatum and Kota Jihuang decoction can significantly reduce the amount of serum hepatic heme and the activity of serum alanine aminotransferase, but have no cholagogic effect.

3. antibacterial and antiviral effects: emodin and 7-acetyl-2-methoxy-6-methyl-8-hydroxy-1,4-naphthoquinone have antibacterial activity. The in vitro antibacterial test of resveratrol showed that it had strong antibacterial properties against the dark red dermatophytes and interdigital dermatophytes that lead to intractable tinea and blister like tinea alba, and had strong bactericidal effects against Bacillus subtilis and Xanthomonas gambogi. Emodin PD、 Emodin-8-glucoside has inhibitory effect on Staphylococcus aureus and diplococcus hepaticus. A flavonoid contained in Polygonum cuspidatum has inhibitory effect on Staphylococcus aureus, Staphylococcus albus and Proteus. . The 3% decoction had strong inhibitory effect on seven representative virus strains, including 479 adenovirus type 3, 72 polio type II, 44 echo type 9, coxsackie A9 and B5, encephalitis B (jingweiyan strain I), 140 herpes simplex. Polygonum cuspidatum monomer Ⅰ and Ⅱ can reduce the titer of hepatitis B antigen by 8 times.

4. antitussive and antiasthmatic effects: experiments using electrical stimulation of the superior laryngeal nerve in cats showed that emodin PD、 Compound Yin Yang Lotus (Polygonum cuspidatum, Mahonia Mahoniae, and loquat leaf Decoction all have antitussive effects. The constant pressure ammonia spray method in mice also showed that PD has antitussive effects. . It has no antagonistic effect on acetylcholine induced tracheal contraction.

5. effect on the release of prostacyclin (PGI2) and morphology of cultured human umbilical vein endothelial cells: the effect of PD on the release of prostacyclin from primary cultured human umbilical vein endothelial cells was studied by measuring the concentration of 6-Keto-PGF1a in the culture solution with radioimmunoassay. The results showed that PD increased the release of PGI2 in a dose-dependent manner. When pd0.46.0.15mmol / l was applied for 10 minutes, 6-Keto-PGF1a increased significantly, and the effect of the former was still very significant until 30 minutes, but it did not increase when the dose was lower than 0.15mmol / L. .

6. antitumor effect: anthraquinones can inhibit the synthesis of DNA and RNA in human promyelocytic leukocyte (HL-60) cells. The mechanism of action is mainly to inhibit the synthesis of DNA and RNA. The cytotoxic effect of emodin is the strongest. At the concentration of 100umol / L (about 40 μ g/ml), the order of inhibitory effect on the incorporation of DNA precursor [3H me] dthd is: emodin, emodin-8-o-d-glucoside > emodin-8-o-d-glucoside > Rhein > polydatin. The inhibitory effect on the incorporation of [5.3h] CYD, an RNA precursor, was smaller, and the order was: emodin > emodin methyl ether > emodin methyl ether-8-o-d-glucoside, polydatin. Emodin has stronger cytotoxic effect than its glucoside, while other components have no such structure-activity relationship. After oral administration of Polygonum cuspidatum Decoction for 10 days, the tumor inhibition rate of Ehrlich ascites cancer in mice was 35.3%, and the survival time of animals could be prolonged. Emodin inhibited more than 30% of seven tumor strains, including mouse sarcoma, mouse liver tumor, mouse breast cancer, mouse Ehrlich ascites cancer, mouse lymphosarcoma, mouse melanoma, and white rat Walker cancer.

7. hypoglycemic effect: the oxalic acid extracted from Polygonum cuspidatum by intravenous injection can cause hypoglycemic shock. Polygonum cuspidatum can reduce the incidence and mortality of experimental animal diabetes.

8. hypolipidemic effect: Resveratrol glycoside can significantly reduce serum cholesterol in normal rats by gavage of 200mg / kg for 7 consecutive days, while Polygonum cuspidatum decoction has no obvious effect, which may be caused by the low content of resveratrol in the decoction.

9. hemostatic effect: Polygonum cuspidatum decoction has obvious hemostatic effect on traumatic bleeding, and oral administration also has hemostatic effect on upper gastrointestinal bleeding.

10. other effects: Polygonum cuspidatum extract has antipyretic and analgesic effects. Resveratrol glycoside has synergistic effect with sodium pentobarbital and ethyl carbamate, and can significantly prolong the sleep time of mice. Polygonum cuspidatum decoction has convergence, prevention of infection and anti-inflammatory effects on scalded wounds. In addition, a certain concentration of emodin can cause the increase of small intestinal muscle tension and contraction amplitude, and increasing the dose can inhibit the activity of small intestine.