Wattle bark
1、 Pharmacopoeia standard of Vitex negundo       This product is the dried root bark or near root bark of Pseudolarix amabilis (Nelson) rehd., a Pinaceae plant. .
      [character] the root bark is irregular long strip, twisted and slightly curled, with different sizes and thicknesses of 2-5mm. The outer surface is grayish yellow and rough, with wrinkles and grayish white transverse lenticel like protrusions. The thick bark is often scaly exfoliated, and the exfoliation is reddish brown; The inner surface is yellowish brown to reddish brown, flat, and has fine longitudinal texture. It is tough, and the fracture surface is split and can be peeled off layer by layer. .
      The bark is lamellar, about 8mm thick, and thick. .
      [identification] (1) the powder of this product is light brown or brownish red. There are many stone cells, which are rectangular, round or irregular branched, with a diameter of 30-96 μ m, and contain yellow brown patches. The sieve cells are mostly in bundles, with a diameter of 20-40 μ m. There are many oval sieve domains on the side wall. . The resin cells are longitudinally connected into tubes, containing reddish brown to yellowish brown resin like substances, and some are buried with calcium oxalate crystals. The cork cell wall is slightly thick, some are lignified and have pits.
      (2) Take LG of the product powder, add 20ml of methanol, sonicate for 20 minutes, cool, filter, and take the filtrate as the test solution. In addition, LG, a control medicinal material from the bark of Vitex negundo, was prepared into a control medicinal material solution by the same method. Then take the acetic acid reference substance of Vitex negundo bark and add methanol to make a solution containing 0.2mg per 1ml as the reference solution. According to the test of thin layer chromatography (general rule 0502), suck 5 μ l of each of the above three solutions, dot them on the same silica gel G thin layer plate respectively, use toluene ethyl acetate formic acid (14:4:0.5) as the developing agent, develop, take out, air dry, spray 10% sulfuric acid ethanol solution, heat at 105 ℃ until the spots develop clearly, and check under the ultraviolet light (365nm). In the chromatogram of the test sample, fluorescent spots of the same color are displayed at the corresponding positions of the chromatogram of the control medicinal material and the chromatogram of the control sample.
      [inspection] the moisture content shall not exceed 13.0% (the second method of general rule 0832).
      The total ash content shall not exceed 6.0% (general rule 2302).
      Acid insoluble ash content shall not exceed 2.0% (general rule 2302).
      [extract] according to the hot leaching method under the alcohol soluble extract determination method (general rule 2201), 75% ethanol shall be used as solvent, which shall not be less than 15.0%.
      [content determination] determine according to HPLC (general rule 0512).
      Chromatographic conditions and system suitability test octane silane bonded silica gel was used as filler; The mobile phase was methanol-1% acetic acid solution (50:50); The detection wavelength is 260nm. The number of theoretical plates should not be less than 5000 according to the acetic acid peak of Vitex negundo bark.
      Preparation of reference solution take an appropriate amount of wattle bark acetic acid reference substance, weigh accurately, add methanol to make a solution containing 45 μ g per 1ml.
      Preparation of test solution take about 0.2g of the powder (passing through No. 3 screen), accurately weigh it, place it in a corked conical flask, precisely add 25ml of methanol, close the stopper, weigh it, heat and reflux for 1 hour, cool it, weigh it again, make up the lost weight with methanol, shake it well, filter it, 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.
      According to the calculation of dry product, the content of wattle bark acetic acid (c23h29o8) shall not be less than 0.25%.
      Decoction pieces
      [processing] wash, slightly moisten, shred and dry.
      This product is in strip or roll shape. . The inner surface is yellowish brown to reddish brown, with fine longitudinal lines. The cut surface is reddish brown to reddish brown, and sometimes small white crystals can be seen, which can be peeled off layer by layer. .
      [inspection] the total ash content is the same as that of medicinal materials, and shall not exceed 5.0%.
      [identification] [inspection] (water acid insoluble ash) [extract] [content determination] is the same as that of medicinal materials.
      [nature, taste and meridian tropism] pungent, warm; Toxic. Return to the lung and spleen meridians.
      [functions and indications] it can kill insects, treat ringworm and relieve itching. For itching and itching.
      [usage and dosage] Apply appropriate amount externally, dip and rub with vinegar or wine, or mix and coat the affected part with grinding powder.
      [storage] put it in a dry place.
 
2、 Chemical constituents of Vitex negundo bark
    Root bark containsPseudolaricacid a、pseudolaric acid B、pseudolaric acid C、pseudolaric acid D、pseudolaric acid E， The two kinds of acid C2 are demethylpseudolaricacid B, pseudolaricacida - β - d-glu-coside, pseudolaricacidb- β - D-glucoside, pseudolaricacib- β - D-glucoside, pseudolarifuroic acid, betulinicacid, β - sitosterol, β - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -s-t-t-t-t-t-t-t-t-t-t-t-t-t-t-t-t-t-t-t-t-t-t-t-t-t-t-t-t-t-t-t-t-t-t-t-t-t-t-t-t. The seeds contained pseudolarolide e, h, and pseudolarolide a, B, C, D, and I. Tree rings contain lead, iron, calcium, manganese and zinc.


3、
1. antifungal effect. Take 0.2cm of psoriatic ringworm colony and transplant it on sabourand's glucose agar medium. After 3wk, cut 0.2cm diameter colonies and inoculate them in 40% Vitex bark liquid medium and control medium. Inoculate three tubes each, incubate at 25 ℃ for 2h, and then make ultrathin sections according to the conventional treatment. The observation by electron microscope shows that Vitex bark has a strong antifungal effect. The degree of cell damage is related to the duration of drug action when the drug concentration is the same as Liu Liang 's. if the fungus is fully exposed, when the drug concentration is sufficient, it has 100% cell killing effect. Compared with the effects of antifungal agents such as amphotericin B, the same point is that there are thin electronic areas in the cytoplasm, the cell membrane is intact, and it is reflexed into the cytoplasm. The difference lies in the degeneration of the fungal cell structure under the action of Vitex negundo, the disappearance of organelles, the obvious shrinkage of the cell membrane, and the diverticulum like reverse folding into the cell head, and the degeneration or even disappearance of the mitochondrial structure. The main antifungal components in the bark of Vitex negundo are Vitex formic acid, Vitex acetic acid and Vitex propionic acid.

2. the effect of Vitexin on human liver cancer cells SMMC-7721 was cultured in RPMI-1640 medium containing 30% calf serum. The dense monolayer cells were made into county liquid and inoculated in culture flasks with the same number of cells. The cells were cultured at 37 ℃ for 2 days for experimental use, and the human diploid cells 2BS were used as the control. Vitexin was made into injection (20mg/ml) with 3.5% Tween 80. In the experiment, the above culture solution was used to prepare three groups of Vitexin with concentrations of 10ug, 5ug and 265ug/ml, which were respectively added to the culture bottle with prepared cells for further culture. The total number of cells and the number of dead cells in each group were counted to calculate the killing rate of the drug on cells. The results showed that compared with the control group, most of the tumor cells in the 10 and 5ug/ml groups became round, the cell membrane became thick and wrinkled, and both fell off in suspension, and only a few cells adhered to the wall; 10. ; The inhibition rate of protein content increased with the increasing concentration of Vitexin in each group. When the dose of Vitexin was 10ug/ml, the inhibition rate was 64.54%; When the concentration of Vitexin was 10, 5 and 2.5ug/ml, the cell killing rates were 42.9%, 39.3% and 29.4%, respectively, and only 8.3% of the cells in the control group died.

3. anti fertility effect
  . For pregnant hamsters, 60 mg / kg of trifolic acid had a very significant anti early pregnancy effect compared with the control group (P < 0.01 =), while 20-40 mg / kg of trifolic acid had a significant anti early pregnancy effect (P < 0.05 or P < 0.01 =). The anti early pregnancy effect of Vitex trifolia acetic acid 30-40mg/kgqdx3d given to rabbit SC and IV solution was better than that of Ig suspension 40mg/kgqdx3d. . Two dogs were given a single dose of Ig suspension of 20-30mg/kg, and all the fetuses also died. The anti early pregnancy effect of terrestris acetic acid may first damage the decidual part of the mother, and the decidual necrosis is not due to the decline of progesterone. Experiments showed that exogenous progesterone could not resist its anti early pregnancy effect.

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