1、 Pharmacopoeia standard of Sophora flavescens
This product is the dried root of Sophora flavescens ait., a legume plant. Excavate in spring and autumn, remove root heads and small branch roots, wash and dry, or slice while fresh and dry.
[properties] this product is a long cylindrical shape with branches at the lower part, 10-30cm long and 1-6.5cm in diameter. Its surface is grayish brown or brownish yellow, with longitudinal wrinkles and transverse lenticel like protrusions. Its outer skin is thin, often broken and rolled, easy to peel off, and the peeling part is yellow and smooth. Hard, not easy to break, fibrous section; . The breath is slight and the taste is extremely bitter.
. ; The surface appears to be quasi polygonal, with irregular fine cracks on the surface of the flat circumferential wall and intermittent pits on the vertical circumferential wall. Fibers and crystalline fibers, mostly in bundles; The fiber is slender, 11-27 µ m in diameter, with thick wall and non lignified; The cells around the fiber bundles contain calcium oxalate crystals, forming crystal fibers, and the walls of crystal containing cells are unevenly thickened. Calcium oxalate square crystals are biconical, rhombic or polyhedral in shape, with a diameter of about 237 µ M. starch grains, single round or oblong in shape, with a diameter of 2-20 µ m, umbilical point crack like, and large grain lamina striae are faintly visible; There are many complex grains, which are composed of 2-12 grains.
(2) Take a cross section of this product, add a few drops of sodium hydroxide test solution, and the cork will turn orange red, gradually change to blood red, and will not disappear after a long time. Xylem did not show color reaction.
(3) Take 0.5g of this product powder, add 0.3ml of concentrated ammonia test solution and 25ml of chloroform, place it overnight, filter, evaporate the filtrate, add 0.5ml of chloroform to dissolve the residue, and use it as the test solution. In addition, take matrine reference substance and sophoridine reference substance, add ethanol to make a mixed solution containing 0.2mg per 1ml as the reference solution. . In the chromatogram of the test sample, the same orange spots appear at the corresponding positions of the chromatogram of the control sample.
(4) . Test according to thin-layer chromatography (general rule 0502), suck 4 µ l each of the test solution and the above reference solution under item (3) of [identification], dot them on the same silica gel G thin-layer plate prepared with 2% sodium hydroxide solution, use the lower solution of chloroform methanol concentrated ammonia test solution (5:0.6:0.3) placed below 10 ℃ as the developing agent, develop, take out, air dry, and spray bismuth potassium iodide test solution and sodium nitrite ethanol test solution in turn. In the chromatogram of the test sample, the same orange spots appear at the corresponding positions of the chromatogram of the control sample.
[inspection] the moisture content shall not exceed 11.0% (the second method of general rule 0832).
The total ash content shall not exceed 8.0% (general rule 2302).
[extract] determined according to the cold leaching method under the determination method of water-soluble extract (general rule 2201), it shall not be less than 20.0%.
[content determination] determine according to high performance liquid chromatography (general rule 0512).
Chromatographic conditions and system suitability test amino bonded silica gel was used as filler; The mobile phase was acetonitrile absolute ethanol-3% phosphoric acid solution (80:10:10); The detection wavelength is 220nm. The number of theoretical plates should not be less than 2000 according to the Oxymatrine peak.
Preparation of reference solution take appropriate amount of matrine reference and Oxymatrine reference, weigh accurately, add acetonitrile absolute ethanol (80:20) mixed solution to make a solution containing 50 µ g of matrine and 0.15mg of oxymatrine per 1ml.
Preparation of test solution take about 0.3g of powder (passing through No. 3 screen), weigh accurately, place it in a corked conical flask, add 0.5ml of concentrated ammonia test solution, precisely add 20ml of chloroform, close the stopper, weigh, sonicate (power 250W, frequency 33khz) for 30 minutes, cool, weigh again, make up the lost weight with chloroform, shake well, filter, accurately measure 5ml of secondary filtrate, add it to a neutral alumina column (100-200 mesh, 5g, inner diameter LCM), elute with 20ml of chloroform and chloroform methanol (7:3) mixed solution in turn, and combine and collect the eluent, recover the solvent to dryness, add an appropriate amount of absolute ethanol to dissolve the residue, transfer it to a 10ml volumetric flask, add
Absolute ethanol to the scale, shake well, and get it.
The determination method is to precisely suck 5 µ l of each of the above two reference solutions and 5 ～ 10 µ 1 of the test solution, inject them into the liquid chromatograph, and determine.
The total amount of matrine (c15h24n2o) and Oxymatrine (c15h24n2o2) in this product shall not be less than 1.2% according to the dry product.
Decoction pieces
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This product is a thick piece of quasi circular or irregular shape. The outer skin is grayish brown or brownish yellow. Sometimes transverse lenticel like protrusions can be seen. The outer skin is thin, often broken, rolled back or falling off. The falling off part is yellow or brownish yellow, smooth. The cut surface is yellowish white, fibrous, with radial texture and cracks, and some concentric rings can be seen. The breath is slight and the taste is extremely bitter.
[content determination] the total amount of matrine (c15h24n2o) and Oxymatrine (c15h24n2o2) in the same medicinal material shall not be less than 1.0%.
[identification], [inspection], [extract] are the same as that of medicinal materials.
. Homing, liver, stomach, large intestine, bladder meridian.
[functions and indications] clear away heat and dampness, kill insects and diuresis. It is used for hot dysentery, hematochezia, jaundice, urinary incontinence, erythema and leucorrhea, Yin swelling and Yin itching, eczema, wet sores, skin itching, scabies and leprosy; External treatment of trichomonas vaginitis.
[usage and dosage] 4.5 ～ 9g. Apply appropriate amount externally, and wash the affected area with Decoction.
[note] it should not be used with resveratrol.
[storage] put it in a dry place.

2、 Chemical constituents of Sophora flavescens
  Alkaloids in flowers: d-matrine, d-oxymatrine, l-7,11-dehydromatrine, d-5a, 9a dihydroxymatrine, l-sophora root, n-oxysophora root, l-sophoramine, l-9a-hydroxysophoramine, d-sophoraol, sophoraol-n-oxide, lupanine, l-smelly bean, l-pseudoindigo leaf alkali, levo-n-methylgenistein, levo rhombifoline, d-mamanine, d-kuraranine, isokuraramine.


3、 Pharmacological action of Sophora flavescens

1. antitumor effect: matrine has anticancer activity. The survival rate of mice in the 500 μ g · D intraperitoneal injection group at 2Mo was 40%, while the control group all died within 23 days. Oxymatrine had no effect. Matrine has the effect of reducing the inhibition of P815 tumor cell proliferation by mouse peritoneal macrophages in vitro. . Matrine, Oxymatrine, sophocarpine and their mixed a, B and C alkaloids in different proportions all have different degrees of inhibition on S-180 solid tumor. The tumor inhibition rate of each monomer alkaloid is more than 35%. When the dose of mixed C alkaloids in different proportions is 113mg / (kg · d), the tumor inhibition rate is 61.38% after continuous intraperitoneal injection for 10 days, which is 23.5% higher than the total alkaloids. There is a significant difference compared with Matrine alone (P < 0.01). Matrine has a significant inhibitory effect on the colony generation rate of peripheral multidirectional hematopoietic progenitor cells in chronic myeloid leukemia, the inhibitory rate is 0.76, and the optimal inhibitory concentration is 100mg / L. Sophora flavescens decoction can significantly induce leukemia cells to differentiate into mononuclear macrophages when it acts on human promyelocytic leukemia cells cultured in vitro at a dose of 8mg / ml.

2. leukocyte increasing effect: Oxymatrine can prevent leukopenia in mice caused by MMC and cyclophosphamide. Intravenous or intramuscular injection of 30mg/kg total matrine and 100mg / kg Oxymatrine can significantly increase the number of white blood cells in peripheral blood of normal rabbits. Moreover, compared with total matrine, Oxymatrine has basically the same effect time, maintenance time and peak white blood cell count in normal rabbits within 18 days after administration, and the leukocyte increasing effect is better than that of shark liver alcohol. In the treatment experiment of total matrine, Oxymatrine and matrine on leukopenia caused by 600 X-ray irradiation in rabbits, the white blood cell count in the total matrine group remained above 6000 / mm3 from 5 days after administration to day L9, while the control group did not recover the above level until 22 days; The whitening potency of oxymatrine was not higher than that of total alkaloids, while matrine had no therapeutic effect. Oxymatrine has a certain preventive and therapeutic effect on leukopenia induced by 500 cycles of 60 Co r-ray total body irradiation in rabbits, with a dose rate of 31 cycles / min, but no preventive and therapeutic effect at a dose rate of 120 cycles / min.

3. effects on cardiovascular system: intravenous injection of matrine and Oxymatrine with 1/4ld50 significantly antagonized arrhythmias induced by aconitine and chloroform epinephrine in rats; Intraperitoneal injection significantly antagonized chloroform induced ventricular fibrillation in mice, and also increased the dosage required for aconitine induced arrhythmia in rats, and antagonized barium chloride induced arrhythmia in rats and arrhythmia induced by ligation of the anterior descending coronary artery in rats. 30mg / kg Oxymatrine was intravenously injected into rabbits to shorten the recovery time of arrhythmia induced by epinephrine, with an average reduction of 43% for treatment and 22% for prevention. ; Intravenous injection of l6mg / kg can increase the dosage of ouabain induced premature beats and cardiac arrest in rabbits, and can also resist coronary artery occlusion reperfusion induced arrhythmias in dogs, but it can not resist calcium chloride acetylcholine induced atrial fibrillation (flutter) in mice and chloroform epinephrine induced arrhythmias in rabbits. Intravenous injection of sophoridine (11mg / kg) with 1 / 5 LD50 can prevent or treat aconitine induced arrhythmia in rats, and can also improve the tolerance of guinea pigs to ouabain induced arrhythmias, and prevent arrhythmias induced by barium chloride and calcium chloride in rats. 1 / 10 LD50 dose (5mg/kg) significantly shortened the time of arrhythmia induced by epinephrine in rabbits, and the duration of action was much longer than that of quinidine. . Sophora alkaloids can dilate blood vessels and protect against acute myocardial ischemia. It can cause obvious hypotensive effect on anesthetized rabbits. Generally, the hypotensive effect is about 20mmhg, lasting for 2-3 minutes and returning to normal; The isolated rabbit ear blood vessels can be immediately dilated and last for about 10 minutes; It can prolong the arrest time of acute hemorrhagic heart for about 7 minutes; It has obvious protective effect on acute myocardial ischemia caused by pituitrin. Oxymatrine can increase atrial contractility in rabbits with a good dose-response relationship. 9 μ m does not affect myocardial excitability, but shortens the functional refractory period; Reduce the threshold concentration of adrenaline induced left atrial automaticity; Slow down the automatic rhythm of the right atrium and reduce the positive frequency effect of CaCl2 on the right atrium. The toxic dose (360 μ m) can reduce myocardial contractility and excitability. Oxymatrine 50 μ mol / l could antagonize the positive frequency effect of isoproterenol. . Matrine could slow down the automatic frequency of the right atrium, increase the contractility of the right atrium and reduce the MDF (maximum driving frequency) of the left atrium in a dose-dependent manner; Its negative frequency, positive muscle strength and negative MDF effect are all linearly correlated. Matrine can inhibit aconitine induced left atrial autorhythm in rats or prolong the latency and slow down the initial frequency of aconitine induced autorhythm; . Matrine could be significantly inhibited by the Ca2 + channel blocker verapamil (1 μ mol / L), and the inhibition of smooth muscle was greater than that of myocardium, suggesting that the inotropic effect of matrine may be related to the activation of Ca2 + channels. Matrine can increase cAMP content and reduce cAMP level in rat plasma, but has no significant effect on camp and cGMP content in myocardial tissue. Sophoridine can increase cAMP levels in plasma and myocardial tissue of rats. Oxymatrine 50 μ mol / L and 250 μ mol / L significantly inhibited the increase of lactate dehydrogenase content in cultured cardiomyocytes induced by mitomycin C and alleviated cardiomyocyte injury caused by glucose deprivation and hypoxia, but had no protective effect on cardiomyocyte injury caused by chlorpromazine. Intramuscular injection of oxymatrine can significantly prolong the survival period of free allografted myocardium in mice, and its effect increases with increasing dose. Total flavonoids of Sophora flavescens at 125-250 μ g / ml could slow down the frequency of spontaneous beats of cultured neonatal rat cardiomyocytes and antagonize the spontaneous and ouabain induced arrhythmias of cardiomyocytes. The total flavonoids of Sophora flavescens had obvious antagonistic effect on ventricular fibrillation induced by chloroform inhalation in mice, with LD50 of 23.9 ± 1.1g / kg and therapeutic index of 3.56. After intravenous injection of 8G and 30g / kg of total flavonoids of Sophora flavescens into rabbits for 30-60 minutes, it can obviously resist arrhythmia caused by chloroform adrenaline. The complete confrontation rates were 40% and 70%, respectively. The total flavonoids of Sophora flavescens (20-40g / kg) injected intravenously has obvious therapeutic effect on arrhythmia induced by intravenous aconitine in rats. The effective rate of the first treatment was 63%, and the effective rate of the second treatment was 84%. The intravenous injection of matrine in rats can significantly antagonize the arrhythmia caused by aconitine, barium chloride and ligation of coronary artery. The intravenous injection of total flavonoids of matrine 30g / kg and 60g / kg in anesthetized rats showed obvious negative autonomic effect, negative frequency effect and negative conduction effect. .

. Matrine can antagonize the effects of acetylcholine and barium chloride on isolated guinea pig and rat trachea and intestine in the presence or absence of calcium Krebs nutrient solution. The antiasthmatic effects of matrine and Oxymatrine on histamine induced asthmatic guinea pigs were basically similar to aminophylline. The antiasthmatic rate of 1 hour was more than 90%, while that of the control group was about 15%; In terms of antiasthmatic time, the antiasthmatic rate of aminophylline in 6 hours was 55%, while the total alkaloid of Sophora flavescens was more than 80%. The asthmatic effect of sophocarpine ACh is stronger than aminophylline, and its antiasthmatic effect may be through the excitation of β - receptors in the midbrain. Phenol red excretion method proved that mice were gavaged with 0.8g/kg total alkaloids and flavonoids of Sophora flavescens, which had obvious dispelling effect.

5. sedative effect: matrine 50-100mg / kg can significantly inhibit the free activity of mice; 400mg / kg significantly inhibited the passive activity of mice; ; Matrine 25-40mg / kg combined with dongmianling (5mg / kg) can cause the righting reflex of mice to disappear. Matrine 100-200mg / kg can significantly enhance the inhibitory effect of reserpine 0.5mg / kg on spontaneous activity of mice; The combination of 50-200mg / kg matrine and 20-25mg / kg pentobarbital sodium at subthreshold dose by intraperitoneal injection can cause mice to fall asleep; Combined with thiopental sodium (intraperitoneal injection, 40mg / kg) and chloral hydrate (intraperitoneal injection, 200mg/kg), respectively, can significantly enhance the central inhibition. It can also significantly resist psychomotor excitation induced by amphetamine (intraperitoneal injection, 4-6mg / kg) and caffeine (50mg / kg), which are central stimulants; The total alkaloids of Sophora flavescens not only had no antagonistic effect on the convulsion caused by strychnine and pentylenetetrazol, but also enhanced the convulsion and increased the number of animal deaths; The total alkaloid of Sophora flavescens has mild analgesic effect alone, and its analgesic percentage can be significantly increased when combined with threshold dose morphine.

6. anti allergic effect: matrine can reduce the release of allergic mediators and is an immunosuppressant. Its concentration of inhibiting 50% T cell proliferation is 0.55-0.56mg / ml; The concentration of inhibiting IL-2 production was 0.1mg / kg.

7. immunosuppressive effect: matrine and Oxymatrine can inhibit the immune function of mice at the dose of 1 / 5 LD50. The intramuscular injection of oxymatrine 150mg / kg or 100mg / kg in rabbits and rats significantly inhibited the passive or active cutaneous anaphylaxis and the formation of serum IgE antibody in rabbits. Intraperitoneal injection of oxymatrine 200mg / kg · D for 21 days had a protective effect on the death caused by rapid allergic reaction in mice.

8. other effects: intramuscular injection of oxymatrine can significantly resist the exudative inflammation induced by croton oil, carrageenan (rats) and glacial acetic acid (mice). Rabbits were treated by gavage, subcutaneous injection, intravenous injection, intraperitoneal injection and intramuscular injection of Sophora flavescens decoction, Sophora flavescens injection and matrine, and the amount of sodium chloride in urine was significantly increased. Sophora flavescens 50% methanol extract has anti ulcer effect caused by hydrochloric acid and ethanol, and its active ingredient is kurarinone. In addition, matrine has antibacterial effect both in vitro and in vivo, and its intensity in vivo is equivalent to that of chloramphenicol. The alcohol extract has anti Trichomonas effect in vitro, and its intensity is similar to that of hemp seed.