Agrimonia pilosa
1、 Pharmacopoeia standard of Agrimonia pilosa       This product is the dry aboveground part of Agrimonia pilosa ledeb., a Rosaceae plant. When the stems and leaves are luxuriant in summer and autumn, they are cut to remove impurities and dried.
      [character] the product is 50 ~ 100cm long and is covered with white fluff. The lower part of the stem is cylindrical, with a diameter of 4-6mm, reddish brown, and the upper part is square columnar, slightly concave on all sides, greenish brown, with longitudinal grooves, prismatic lines, and knots; Light, hard, easy to break, hollow section. Odd pinnate compound leaves alternate, dark green, wrinkled and curly; ; There are two kinds of leaves in size, alternating on the leaf axis, the top leaflet is larger, the complete leaflet is oval or long oval after flattening, the apex is pointed, the base is cuneate, and the edge is serrated; Stipules 2, amplexicaul, obliquely ovate. The raceme is slender, the lower part of the calyx is tubular, the upper part of the calyx tube has barbs, the apex is 5-lobed, and the petals are yellow. The breath is slight, and the taste is slightly bitter.
      [identification] (1) the powder of the leaves of this product is dark green. The upper epidermal cells were polygonal; The cell wall of the lower epidermis was unduly curved, and the stomata were indeterminate or unequal. . The glandular hair head is 1-4 cells, oval, and the stalk is 1-2 cells; There are also a few glandular scales with single cells in the head, about 68 µ m in diameter, oil droplets, and single cells in the stalk. There are many calcium oxalate clusters with a diameter of 9-50 µ M.
      (2) . Add 10ml of trichloromethane to dissolve the residue, shake and extract with 10ml of 5% sodium hydroxide solution, discard the trichloromethane solution, adjust the pH value of the sodium hydroxide solution to 1-2 with dilute hydrochloric acid, shake and extract with trichloromethane twice, 10ml each time, combine the trichloromethane solution, add 10ml of water to wash, discard the water solution, and concentrate the trichloromethane solution to 1ml as the test solution. Another 2G of Agrimonia pilosa reference medicinal material was prepared into the reference medicinal material solution by the same method. Then take agrimonic phenol B reference substance and add chloroform to make a solution containing 0.5mg per 1ml as the reference solution. According to the test of thin-layer chromatography (general rule 0502), suck 10 µ l of each of the above three solutions, dot them on the same silica gel G thin-layer plate, use the upper solution of petroleum ether (60 ～ 90 ℃) - ethyl acetate acetic acid (100:9:5) as the developing agent, develop, take out, dry, spray 10% ethyl sulfate solution, heat at 105 ℃ until the spots are clear. .
      [check] moisture   .
      Total ash   Not more than 10.0% (general rule 2302).
      Decoction pieces
      [processing] remove residual roots and impurities, wash, moisten slightly, cut into sections, and dry.
      This product has irregular segments, most of the stems are square columnar, with longitudinal grooves, prismatic lines and knots. The cut surface is hollow. The leaves are mostly broken, dark green, with serrated edges; . Sometimes yellow flowers or fruits with barbs can be seen. The breath is slight, and the taste is slightly bitter.
      [check] moisture   .
      [identification] the same as medicinal materials.
      [nature, taste and meridian tropism] bitter, astringent and flat. Heart returning and liver meridian.
      [functions and indications] astringent and hemostatic, stop malaria, stop dysentery, detoxify and tonify deficiency. .
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2、 Chemical constituents of Agrimonia pilosa l.var.pubescens l.var.pubescens l.var.pubescens l.var.pubescens l.var.pubescens l.var.pubescens l.var.pubescen
The whole grass contains agrimoninAgrimonine(Agrimonolide)、 Tannins (pyrocatechol tannins, gallotannins, etc. (6.67% - 13.71%)), sterols, organic acids, phenolic components, saponins, agrimola, agrimola, agrimola, agrimola, agrimola, agrimola, agrimola, agrimola, agrimola, agrimola, agrimola, agrimola, agrimola, agrimola, agrimola, agrimola, agrimola, agrimolaAgrimonic phenol B(agrimolB)、 Agrimonil D(agrimolD)、 Agrimol e, agrimol F and agrimol g. the structure of agrimol C has been clarified; . The content of tannin in roots, stems and leaves was 8.9%, 6.5% and 16.4%, respectively. . Fresh rhizomes and winter buds contain agrimophol, which is proved to be an effective substance by in vitro cysticercosis killing test and clinical observation. It also contains agrimonolide, vanillicacid, l-dihydroquercetin, l-taxifolin)、 Ellagic acid, pseudo mianmarin, agrimonine-6-o - β - D-glucoside, trans-p-hydroxycinnamate, (2S, 3S) - (-) - Douglas fir-3-o - β - D-glucoside, ellagic acid-4-o - β - D-xyloside and Potentilla acid, etc.

3、 Pharmacological effects of Agrimonia pilosa
1. insecticidal effect:
  1.1. drive and kill effect on Schistosoma:

  1.2. killing effect on tapeworm and Ascaris lumbricoides:

2. spermicidal effect:
  2.1. effect of petroleum ether extract from Agrimonia pilosa root and bud on animal sperm: the testes of 6 mice and 4 guinea pigs were used for the experiment. The sperm of two animals were used with two concentrations of petroleum ether extract. The sperm of mice were used for five experiments, and the sperm of guinea pigs were used for three experiments. . Petroleum ether extract 3.1 × 10 (-4) g/ml could also kill all sperm of guinea pigs within 5 minutes, while the concentration doubled, and 6.2 × 10 (-4) g/ml could kill all sperm of guinea pigs within 1 minute.

  2.2. effect of Agrimonia pilosa petroleum ether extract on human sperm: semen from two people was used for the test, and diluted semen was used for some tests, and original semen was used for some tests, and two tests were conducted respectively. Results the petroleum ether extract suspension containing 5 × 10 (-4) g/ml oridonin could kill all sperm in 3-7 minutes; The concentration of oridonin 2.5 × 10 (-3) g/ml can kill all sperm in 1-5 minutes.

  2.3. effect of oridonin suspension on animal sperm: dissolve 1 part of oridonin crystal in 10 parts of chloroform, add it to 50 parts of polyethylene glycol (peg300) in several times, grind it evenly, then add 5% gum arabic solution prepared with Ren's solution, add it while grinding, and prepare the suspension with the required concentration. The preparation of the control solution is the same as that of the crane phenol suspension, except that there is no crane phenol. Different concentrations of oridonin suspension and control solution were diluted with Ren's solution. The testes of 5 mice, 2 guinea pigs and 2 rabbits were used for the test. The sperm of the three animals were tested with 2-3 concentrations of oridonin suspension for 2-4 times. The concentration of 1 × 10 (-4) and 2 × 10 (-4) g / ml of oridonin suspension could kill all sperm of mice, guinea pigs and rabbits within 5 minutes; 3 × 10 (-4) and 5 × 10 (-4) g/ml could kill the sperm of three animals in 1 minute.

  2.4. effect of chrysophanol suspension on human sperm: semen from three people was used for the test, and diluted semen was used for some tests, and raw semen was used for some tests. Four concentrations of oridonin suspension were used for diluted semen test, and two concentrations of oridonin suspension were used for original semen test. Each concentration was tested for 3-5 times respectively. The concentration of chrysophanol suspension at 2 × 10 (-4) and 3 × 10 (-4) g/ml can kill all sperm in 5 minutes, and 4 × 10 (-4) and 5 × 10 (-4) g/ml can kill all sperm in 1 minute; However, for human raw semen, a higher concentration is required. 1 × 10 (-3) g/ml can kill all sperm in 5 minutes, and 2 × 10 (-3) g/ml can kill all sperm in 1 minute.

3. anti tumor effect:
  3.1. effect on cancer cells: take the ascites fluid of liver cancer ascites cancer mice that have been inoculated for 6-9 days and grow well, dilute it with sterile normal saline to 1 × 10 (7) -2 × 10 (7) cancer cells / ml, take 0.5ml of this cancer cell fluid and add the same volume of different concentrations of camphor to make the final concentration of camphor 2.5-0.08mg / ml four concentrations, shake and mix gently, incubate at 37 ℃ for 3 hours, then take it out, add 0.05% eosin, microscopic examination, count the percentage of cancer cells stained red (dead cancer cells are red stained, and the percentage of red staining is called the red staining rate), and the results of four experiments with incubation for 1 h and 3 h are listed in table. The results showed that 0.16mg / ml oridonin had certain killing effect on cancer cells, and 1.25mg / ml could kill all cancer cells within 1 hour. The red staining rate of cancer cells in the normal saline control group within 1 hour was 7.3%. Each mouse was subcutaneously inoculated with 0.2ml of 10 (2) -3 × 10 (2) cancer cells / ml under the right armpit, and each large group of animals was inoculated with a tumor source. The next day, each large group of animals was randomly divided into the administration group and the normal saline control group, and began to intraperitoneally inject 30mg / kg of oridonin injection every day. The control group was injected with the same volume of normal saline for 12-14 days, and the animal weight and tumor weight were weighed the next day after drug withdrawal. The results of two repeated experiments proved that oridonin had obvious experimental therapeutic effect on S37 and U14, and the inhibition rates of tumor growth were 47.0% (p< 0.01) and 38.7% (p< 0.05). It also had certain inhibitory effect on S180 and hepatoma sarcoma, and the inhibitory rates were 26.3% and 23.5%, respectively (p< 0.05). The animals in the administration group did not die during the experiment. After anatomical observation. There was no significant difference in the size of thymus and spleen between the administration group and the control group. The experimental animals of the above four tumor spectra had significantly increased body weight after the experiment. The growth rate of body weight of the animals in the administration group and the control group was similar. It can be seen that this dose of oridonin did not show obvious toxic response to the experimental mice. The inhibition rate of jtc-26 was 100%. . At the concentration of 500 μ g/ml, it not only does not damage normal cells, but also promotes the growth and development of 100% of normal cells (in fact, it has a righting effect).

3.2. effect of local and intraperitoneal administration of tumor on solid tumors: following the above inoculation method, mice were inoculated with S180 and 37, and 30mg / kg of oridonin injection was injected intraperitoneally the next day after inoculation. Animals in the control group were injected with the same volume of normal saline. From the fifth day after administration, the total amount of oridonin injection for animals in the administration group was still 30mg / kg, but each animal was injected with 0.25mg of oridonin the tumor body, and the rest was injected intraperitoneally, starting from the next day after inoculation, for a total of 14 days. On the 15th day, the body weight and tumor weight were weighed, and the results showed that the inhibitory rate of thymol on S180 was 67.7% (p< 0.01). The inhibition rate of S37 was 68.1% (p< 0.01). It can be seen that the curative effect of this mode of administration is more obvious than that of intraperitoneal administration alone.

3.3. effect on ascites cancer: the ascites fluid of liver cancer ascites cancer was extracted, diluted into 1 × 1 (2) -2 × 10 (2) cancer cells / ml with sterile normal saline, and inoculated into mice (each mouse was injected intraperitoneally with 0.2ml). The next day, the mice were randomly divided into the administration group and the control group, and were intraperitoneally injected with oridonin 30mg / kg, once a day, for 7 consecutive days. The control group was injected with the same volume of normal saline, and the death time of each mouse within 30 days was observed (the immortal was calculated as the survival time of 30 days). The life extension rate was calculated. The average survival time of 24 animals in the liver cancer ascites cancer administration group was 26.2 ± 0.9 days, and the average survival time of 25 animals in the control group was 17.5 ± 1.3 days. The growth elongation rate was 49.6% (p< 0.01); Chrysophanol can significantly prolong the life of animals with ascites carcinoma of liver cancer.

3.4. effect on S180: the experimental method was carried out according to the national in vivo screening procedure for antitumor drugs. The results showed that the inhibitory rates of Agrimonia pilosa alcohol extract (root) on S180 in mice were 57.2% (p< 0.001), 53.2% (p< 0.01) and 56.2% (p< 0.01), respectively, after intraperitoneal injection of 1.5g / kg × 10 and repeated experiments; The tumor inhibition rate was 37.424% after oral administration of 1000mg/kg body weight to guinea pigs transplanted with sarcoma-180 once a day; When mice transplanted with sarcoma-180 were intraperitoneally injected with 100mg/kg body weight, the inhibition rate of hot water extract was 18.5%, and that of ethanol extract was 7.4%, or there was water extract from its roots, water extract from its stems and leaves, and alcohol extract had no anti-tumor effect. It is suggested that the antitumor component of Agrimonia pilosa exists in roots (including root buds) and is an alcohol soluble substance. . In the treatment group, the tumor cell nucleus showed reduced division phase, severe degeneration and necrosis, the cytoplasm was reticular or vacuolated, even transparent, the nuclear membrane thickened, and the nuclear chromatin condensed into coarse granules. In severe cases, the nuclear fragmentation and nuclear pyknosis. Electron microscopy showed that the tumor cells in the control group were the same as those seen under light microscopy. The significant changes of tumor cells in the administration group were in the nucleus, the gap between nuclear membranes expanded, and circular nucleoli appeared, and the nucleus was homogenized. In addition, swelling and cristae disappearance of mitochondria in the cytoplasm, vesiculation and degranulation of the rough endoplasmic reticulum, and depolymerization of polyribosomes were also seen.

3.5. the ethanol extract of the whole herb has a tumor inhibition rate of more than 50% on the subcutaneous type of mouse liver cancer.

3.6. oridonl also has inhibitory effect on sarcoma-37 in mice.

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4.1 according to the early research report, Agrimonia pilosa and its alcohol extract can promote blood coagulation. The crude extract also has the effect of promoting blood coagulation, and has the effect of constricting peripheral blood vessels when frog legs are perfused. After intravenous injection of agrimonin in mice, the bleeding time was shortened by 45%. Intravenous injection in rabbits can greatly shorten the hemagglutination time and increase the number of platelets. In experimental dogs with femoral artery hemorrhage, local application of Agrimonia powder combined with compression has a certain hemostatic effect.

; Agrimonin a, B, C and another phenolic resin acid contained in Agrimonia pilosa have no hemagglutination effect.

5. effect on circulatory system: Agrimonia pilosa ethanol extract can increase blood pressure and excite breathing when injected intravenously into anesthetized rabbits and dogs, while the alcohol extract of its water extract can reduce blood pressure in rabbits. When the rabbit ear and frog hind limb were perfused, the low concentration made the blood vessels contract, and the high concentration made the blood vessels expand. It has also been reported that Agrimonia pilosa extract and its extract have no effect on rabbit ear blood vessels when perfused at low concentration, but also show dilation reaction at high concentration, and can resist the vasoconstriction effect of epinephrine. The extracts and agrimonin had cardiotonic effects on isolated frog hearts; It can increase the heart rate and contraction intensity of frogs and toads, while the ethanol extract of water extract can inhibit the isolated frog heart. Agrimonin can increase the contraction amplitude of isolated rabbit heart at the concentration of 1 ∶ 100000-1 ∶ 5000, and the degree of increase is proportional to the concentration. . The results showed that the small dose could increase the amplitude of cardiac contraction, the medium dose could slightly increase the heart rate and amplitude of cardiac contraction, and the large dose could make the cardiac bradycardia and amplitude of contraction larger.

6. effect on smooth muscle: the ethanol extract of the water extracted part can excite the isolated intestine of rabbits and guinea pigs at low concentration and inhibit it at high concentration. Agrimonine can reduce the contraction amplitude and tension of isolated rabbit intestine, and then stop the intestinal movement in a relaxed state. .

. Its anti-inflammatory effect is due to the astringent effect of tannin that can produce condensed phlobaphene in Agrimonia pilosa.

8. antibacterial and antiviral effects: hot water or ethanol extract has a certain inhibitory effect on Bacillus subtilis and Staphylococcus aureus in the test tube, and a weak inhibitory effect on human tuberculosis. It can kill Paramecium, and the effective ingredient can be dissolved in ether and chloroform. The brown powder obtained from methanol extract of Agrimonia pilosa has inhibitory effect on Gram negative bacteria. . The aqueous extract of agrimoniaeupatoria produced in Europe has inhibitory effect on Mycobacterium tuberculosis in vitro, and it is also effective for those who are resistant to streptomycin and salicylic acid, but it is ineffective for those who are resistant to isoniazid. Ethanol extract has antiviral (Columbia virus) effect in mice.

9. other effects: 100mg / kg of water extract has analgesic effect in rabbits (rabbit dental pulp electrical stimulation method), and 50mg/kg of alcohol and water extract is effective. In addition, agrimonin seems to slightly reduce blood glucose, slightly increase the resistance of red blood cells to hypotonic saline, and slightly reduce the basal metabolism of rats. In addition, winter bud powder has cathartic effect, so it is not necessary to take another cathartic in clinical application.