Evodia rutaecarpa
1、 Pharmacopoeia standard of Evodia rutaecarpa       This product is the dried and nearly mature fruit of Rutaceae plants euodia rutaecarpa (juss.) benth., euodia rutaecarpa (juss.) benth. var. officinalis (Dode) Huang or euodia rutaecarpa (juss.) benth. var. bodinieri (Dode) Huang. From August to November, when the fruit has not yet cracked, cut the fruit branches, dry them in the sun or at low temperature, and remove the impurities such as branches, leaves, and fruit stems.
      [character]   The product is spherical or slightly pentagonal oblate, with a diameter of 2-5mm. The surface is dark yellowish green to brown, rough, and there are many dot like protrusions or concave oil spots. . It is hard and brittle, with 5 ovaries in cross section, and 1 pale yellow seed in each room. The aroma is rich, and the taste is spicy and bitter.
      [identification]   (1) This product is powder brown. The non glandular hairs are 2-6 cells, 140-350 μ m long, with obvious mural warts, and some of the cell cavities contain brownish yellow to brownish red substances. The head of glandular hair is 7-14 cells, oval, often containing yellowish brown inclusions; Stalk 2-5 cells. Calcium oxalate has many clusters with a diameter of 10 ～ 25 μ M; . Stone cells are round or rectangular in shape, with a diameter of 35-70 μ m and a large lumen. Oil chamber debris is sometimes visible, light yellow.
      (2) Take 0.4g of this product powder, add 10ml of ethanol, stand for 30 minutes, sonicate for 30 minutes, filter, and take the filtrate as the test solution. In addition, take the rutaecarpine reference substance and rutaecarpine reference substance, add ethanol to make a solution containing 0.2mg and 1.5mg per 1ml, respectively, as the reference solution. According to the test of thin-layer chromatography (general rule 0502), suck 2 μ l of each of the above three solutions, dot them on the same silica gel G thin-layer plate, use petroleum ether (60 ～ 90 ℃) - ethyl acetate triethylamine (7:3:0.1) as the developing agent, develop, take out, dry, and check under the UV light (365nm). In the chromatogram of the test sample, fluorescent spots of the same color appear at the corresponding positions of the chromatogram of the control sample.
      [check]   Impurities shall not exceed 7% (general rule 2301).
      The moisture content shall not exceed 15.0% (the second method of general rule 0832).
      .
      [extract]   According to the hot leaching method under the determination of alcohol soluble extract (general rule 2201), the use of dilute ethanol as solvent shall not be less than 30.0%.
      [content determination]   Determine according to HPLC (general rule 0512)
      ; Octadecylsilane bonded silica gel was used as filler; The mobile phase was [acetonitrile tetrahydrofuran (25:15)] -0.02% phosphoric acid solution (35:65); .
      Preparation of reference solution   Take an appropriate amount of evodiamine reference substance, evodiamine reference substance and limonin reference substance, accurately weigh them, and add methanol to make a mixed solution containing 80 μ g evodiamine, 50 μ g evodiamine and 0.1mg limonin per 1ml.
      Preparation of test solution   Take about 0.3g of this product powder (passing through No. 3 screen), weigh it accurately, place it in a corked conical flask, add 25ml of 70% ethanol precisely, weigh it, soak it for 1 hour, sonicate it (power 300W, frequency 40KHz) for 40 minutes, cool it, weigh it again, make up the lost weight with 70% ethanol, shake it well, filter it, and take the filtrate.
      Assay   Precisely suck 10 μ l of the reference solution and 10 μ l of the test solution respectively, inject them into the liquid chromatograph, and determine.
      The total content of rutaecarpine (c19h17n3o) and rutaecarpine (c18h13n3o) shall not be less than 0.15% and limonin (c26h30o8) shall not be less than 0.20% according to the dry product.
      Decoction pieces
      [processing]   Evodia rutaecarpa to remove impurities.
      ; Same as medicinal materials.
      Prepare Evodia rutaecarpa, mash licorice, add appropriate amount of water, decoct the soup, remove the residue, add pure Evodia rutaecarpa, soak it up, fry it until slightly dry, take it out, and dry it.
      For every 100kg of Evodia rutaecarpa, use 6kg of licorice.
      This product is shaped like Evodia rutaecarpa, and its surface is brown to dark brown.
      [identification] [inspection] (moisture and total ash) [extract] [content determination]   Same as medicinal materials.
      [nature, taste and meridian tropism]   Bitter, bitter, hot; . It belongs to liver, spleen, stomach and kidney channels.
        [function and indications]   It can dispel cold and relieve pain, reduce adverse reaction and stop vomiting, and help yang to stop diarrhea. It is used for Jueyin headache, cold hernia abdominal pain, cold and damp beriberi, menstrual abdominal pain, abdominal distension, vomiting and acid swallowing, diarrhea in the fifth watch.
      [usage and dosage]   2 ～ 5g. Suitable for external use.
      [storage]   Store in a cool and dry place.

2、 Chemical constituents of Evodia rutaecarpa
The seeds contain cis-5,8-tetradecadienoic acid. The volatile oil of fruits contains evodene,Evodia rutaecarpa visfatol（evodol），Limonin（limonin）。 The fruit containsEvodiamine（evodiamine），Rutaecarpine（rutaecarpine），Evodiamine（evocarpine），Hydroxyevodiamine（hydroxyevodiamine）， Wuzhuyin, ocimene, evodione, evogin,Evodiamine（rutaevin）， 7-carboxyevodiamine, dihydrorutaecarpine, 14 formylrutaecarpine, 1-methyl-2-nonyl-4 (1H) - quinolone [1-methyl-2-nonyl-4 (1H) -quinolone], N, n-dimethyl-5-methoxytryptamine, n-methylanthranoylamide), synephrine, dehydroevodiamine, Evodiamide, demethylevodiamide [n- (2-methylaminobenzoyl) tryptamine], 6a-acetoxy-5-epi-limonin, 6 β - acetoxy-5-epilimonin,Berberine（obacunone），Rodanmei lactide（jangomolide）， Rutaevineacetate, graucina, 12a hydroxylimonin, 12a hydroxyevodol, 1-Methyl-2 [(z) - 6-undecylene] - 4 (1H) - quinolone, 1-Methyl-2 [(z) - 6-undecyl]-4 (1H) - quinolone h) - quinolone {1-Methyl-2 - [(z) - 10 pentadecenyl] - 4 (1H) -quinolone}, 1-methyl-2-[(z) - 6-pentadecenyl] - 4 (1H) - quinolone {1-methyl-2-[(z) - 6-pentadecenyl] - 4 (1H) -quinolone}, 1-methyl-2-[(6Z, 9z) - 6, 9-pentadecene] - 4 (1H) quinolone {1-methyl-2-[(6Z, 9z) - 6, 9-pentadecadienyl] - 4 (1H) - quinolone}, 1-methyl-2-[(6Z, 9z) - 6, 9-pentadecadienyl] - 4 (1H) - quinolone},4z7z) - 4,7-tridecadiene]-4 (1H) - quinolone {1-methyl-2-[(4Z, 7z) - 4,7-tridecadienyl]-4 (1H) - quinolone}, Evodia rutaecarpa fruit amide Ⅰ, Fructamide II of Evodia rutaecarpa. In addition, the fruit also containedL-aspartic acid（L-asparticacid），L-tryptophan（L-tryptophan），L-threonine（L-threonine），l-serine （L-serine）， andL-cystine(L-cystine) and other 18 amino acids.

Shihu fruit containsEvodiamide（evodin）,Limonin（limonin），Evodiamine，Rutaecarpine，Hydroxyevodiamine, Shih Hua, evodiamine, dihydroevocarpine, 1-methyl-2-undecyl-4 (1H) - quinolone [1-methyl-2-undecyl-4 (1H) -quinolone], 1-methyl-2-[(z) - 6-undecyl] - 4 (1H) - quinolone {1-methyl-2-[(z) - 6-undecyl] - 4 (1H) - quinolone}. The seeds contain cis-5,8-tetradecadienoic acid. The volatile oil of fruits containsEvodiamine（evodene），Evodia rutaecarpa visfatol（evodol），Limonin（limonin）。 The fruit containsEvodiamine（evodiamine），Rutaecarpine（rutaecarpine）， Evo carpine,Hydroxyevodiamine（hydroxyevodiamine）， Wuzhuyin, ocimene, evodione, evogin,Evodiamine（rutaevin）， 7-carboxyevodiamine, dihydrorutaecarpine, 14 formylrutaecarpine, 1-methyl-2-nonyl-4 (1H) - quinolone [1-methyl-2-nonyl-4 (1H) -quinolone], N, n-dimethyl-5-methoxytryptamine, n-methylanthranoylamide),Synephrine （synephrine），Dehydroaevodiamine（dehydroevodiamine）， Evodiamide, demethylevodiamide [n- (2-methylaminobenzoyl) tryptamine], 6a-acetoxy-5-epi-limonin, 6 β - acetoxy-5-epilimonin,Berberine（obacunone）， Jangomolide, rutaevineacetate, graucina, 12a hydroxylimonin, 12a hydroxyevodol, 1-Methyl-2 [(z) - 6-undecylene] - 4 (1H) - quinolone {1-Methyl-2 [(z) - 6-undecyl]-4 (1H) - quinolone}, 1-Methyl-2 [(z) - 6-undecyl]-4 (1H) - quinolone},z) - 10 pentadecene]-4 (1H) - quinolone {1-methyl-2-[(z) - 10 pentadecenyl] - 4 (1H) -quinolone}, 1-methyl-2-[(z) - 6-pentadecenyl] - 4 (1H) - quinolone {1-methyl-2-[(z) - 6-pentadecenyl] - 4 (1H) -quinolone}, 1-methyl-2-[(6Z, 9z) - 6, 9-pentadecene] - 4 (1H) quinolone {1-methyl-2-[(6Z, 9z) - 6, 9-pentadecadienyl] - 4 (1H) - quinolone}, 1-methyl-2-[(6Z, 9z) - 6, 9-pentadecadienyl] - 4 (1H) - quinolone},4z7z) - 4,7-tridecadiene]-4 (1H) - quinolone {1-methyl-2-[(4Z, 7z) - 4,7-tridecadienyl]-4 (1H) - quinolone}, Evodia rutaecarpa fruit amide Ⅰ, Fructamide II of Evodia rutaecarpa. In addition, the fruit also containedL-aspartic acid（L-asparticacid），L-tryptophan（L-tryptophan），L-threonine（L-threonine），l-serine （L-serine）， andL-cystine(L-cystine) and other 18 amino acids.

Shihu fruit containsEvodiamide（evodin）,Limonin（limonin），Evodiamine，Rutaecarpine，Hydroxyevodiamine, Shih Hua, evodiamine, dihydroevocarpine, 1-methyl-2-undecyl-4 (1H) - quinolone [1-methyl-2-undecyl-4 (1H) -quinolone], 1-methyl-2-[(z) - 6-undecyl] - 4 (1H) - quinolone {1-methyl-2-[(z) - 6-undecyl] - 4 (1H) - quinolone}.

3、 Pharmacological effects of Evodia rutaecarpa
1. effect on cardiovascular system:

1.1. cardiotonic effect:.

1.1.1. the other 8 isolated toad hearts were cross administered with Evodia rutaecarpa Decoction and isoproterenol in the same heart. It is suggested that the cardiotonic effect of Wuzhuyu decoction is small in dose and long in duration, and has no effect of isoproterenol on accelerating heart rate, so it may not increase myocardial oxygen consumption.

1.1.2. three large eared white rabbits, weighing 1.75 ± 0.09kg, of either sex. After anesthesia, open the chest, clamp the apex of the heart with a frog heart clamp, connect the mechanical and electrical transducer, and record the cardiac contraction curve with a two channel physiological instrument. The ear margin vein was given 0.3g/kg Wuzhuyu decoction, and the observation was conducted for 30 minutes. Then, an appropriate amount of 2% Pentobarbital was injected intravenously. After the myocardial contractility was significantly weakened, Wuzhuyu decoction was given, and the observation was conducted for 30 minutes. It can be seen that Wuzhuyu Decoction significantly enhanced the contractility of rabbit hearts in vivo, which did not return to the pre drug level 30 minutes after administration.

1.2. boosting effect:

1.2.1. three dogs were anesthetized, and the left femoral artery blood pressure was displayed with a plug-in sphygmomanometer, and the right femoral vein was given Wuzhuyu Decoction 0.2g / kg. The blood pressure was 13.29 ± 5.71 (kPa, X ± SD, the same below) before administration, and increased after administration, with the highest peak of 24.36 ± 5.60 and the difference of 10.79 ± 1.24 (P < 0.01); The lowest peak was 11.07 ± 5.41, and the difference was 2.20 ± 0.73 (P < 0.05). The pressure rise was maintained at 368 ± 635; There was a slight decrease after recovery, which was maintained for 565 ± 95s.

1.2.2. nine rats were used to observe the effect of 1.04g / kg Wuzhuyu Decoction injection on blood pressure by the right femoral vein with mercury pressure method, and the heart rate was recorded by electrocardiograph. Another 3 rats were used to observe the effect of 0.04mg / kg epinephrine on blood pressure. The results showed that Wuzhuyu decoction had a pressor effect, and the difference was extremely significant before and after administration (P < 0.001). Another 6 rats were selected and divided into two groups. One group was given propranolol 1.8mg / kg in advance, the other group was given phentolamine 3.0mg / kg in advance, and then Wuzhuyu Decoction injection 1.04g / kg after 5 minutes. Results the antihypertensive effect of Wuzhuyu decoction could be abolished by phentolamine pretreatment; . The above results show that Wuzhuyu decoction has a rapid pressor effect, and the pressor effect is accompanied by a slow heart rate. α. The β - receptor blockade experiment showed that wuzhuyuhuang decoction had significant α - excitatory effect and weak β - excitatory effect, and it was a mixed stimulant of α and β receptors. Due to the large human parameters in Wuzhuyu decoction, these experiments cannot fully explain the pharmacological effects of Wuzhuyu on cardiovascular system.

1.3. effects on Bulbar Conjunctival Microcirculation in rabbits.

Eleven white rabbits were fixed on the thermal insulation operation table in the supine position after anesthesia. The bulbar conjunctival microcirculation was observed under the cold light source with the microcirculation microscope magnified by 80 times. The ear marginal vein was slowly injected with 0.3ml / kg of normal saline, and 5 minutes later, Wuzhuyu Decoction injection was injected with 0.3g / kg. The inner diameter of arterioles and the number of capillary network intersections before and after administration were observed and measured, and the dynamic changes of micro blood flow pattern and red blood cell aggregation were recorded. After administration, the micro blood flow velocity increased rapidly; Some micro blood flow patterns were improved, and the particle rheology formed linear particle flow, and the linear particle rheology formed linear flow.

2. effect on digestive system:

2.1. anti gastric ulcer effect:

2.1.1. effect on water immersion stress ulcer: 50 male and female mice weighing 21.2 ± 1.2g were divided into five groups, and fasted for 1 night. After gavage with Evodia rutaecarpa decoction, cimetidine and tap water, they were put into a special stress box and immersed in 20-23 ℃ water to make the water level reach the neck of mice. After 8 hours, the mice were sacrificed, the stomach was removed, and the punctate lesions were examined by microscopy. The number of bleeding points was calculated as the ulcer index. Oral administration of 2G / kg of 50% methanol extract has the anti ulcer effect of water immersion stress in rats, and the inhibition rate is 66.6%.

2.1.2. effect on indomethacin and ethanol induced ulcers: 50 male and female mice weighing 23.4 ± 1.9g were divided into five groups, and were gavaged with Evodia rutaecarpa decoction, furazolidone and tap water respectively after fasting for 24 hours......................................the mice were fed with water decoction of Evodia rutaec. After 30 minutes, indomethacin 10mg / kg was injected intraperitoneally, and then 50% ethanol 0.2ml / mouse was given by gavage 1 hour later. The stomach was sacrificed 1 hour later, and the strip damage was examined by microscopy. The total length (mm) was measured as the ulcer index. Evodia rutaecarpa at 10 and 20g / kg significantly inhibited the ulcer formation induced by indomethacin plus ethanol.

2.1.3. effect on hydrochloric acid-induced gastric ulcer: 40 male and female rats, weighing 176 ± 21g, were divided into four groups. After fasting for 24 hours, they were given Evodia rutaecarpa extract, cimetidine and tap water respectively. After 1 hour, they were given 1ml of 0.6N hydrochloric acid. After another 1 hour, they were sacrificed and the stomach was taken. The strip damage was examined microscopically. The total length (mm) was measured as the ulcer index. The results are shown in Table 8. Evodia rutaecarpa 5 and 10g / kg significantly inhibited the formation of hydrochloric acid ulcer.

2.1.4. effect on ligation of pylorus ulcer: 20 male and female rats weighing 145 ± 16g were divided into two groups equally. After fasting for 24 hours, the pylorus was ligated with ether anesthesia, and Evodia rutaecarpa Decoction and tap water were injected into the duodenum respectively. The stomach was sacrificed 19 hours after suturing the abdominal wall. The gastric ulcer was detected before microscopic examination, and the grading index was calculated. The results are shown in table 8D. It is suggested that Evodia rutaecarpa 10g / kg can only inhibit the tendency of ulcer formation.

2.2. effect on gastric secretion: Twenty male rats weighing 240-300g were randomly divided into two groups. After fasting for 24 hours (free drinking), the administration group was subcutaneously injected with Evodia Decoction injection at the weight of 10G / kg. Two times in total, with an interval of 1 hour. The control group was given equal volume normal saline in the same way. Thirty minutes after the last administration, the animals were anesthetized, and the abdominal wall was sutured after ligation of the pylorus. Postoperative fasting and water deprivation. . The results showed that Wuzhuyu decoction could reduce the secretion of gastric juice and the acidity of gastric juice in rats.

2.3. effects on gastrointestinal motility:

2.3.1. improving the gastric residual rate: 21 healthy mice weighing 18-22g were randomly divided into two groups. According to the method introduced by Zhang et al., the phenol red residual rate in the stomach was used as the index of the discharge of the chamber. The faster the emptying, the less the gastric residual rate. The results showed that Wuzhuyu decoction could significantly improve the gastric residual rate.

2.3.2. inhibit the movement of gastric strips in vitro: prepare the specimens of gastric strips in vitro, maintain them with Taishi solution, and record the curve changes of gastric strips movement with an organ tester in vitro. According to the self comparison before and after administration. Results when the concentration of Evodia rutaecarpa and Evodia decoction reached 5 × 10 (-3) g / ml and 1 × 10 (-2) g / ml, the spontaneous activity of gastric strips was inhibited, that is, the tension and contraction amplitude were decreased, and the activity could be recovered after washing with the maintenance solution. The effects of Codonopsis pilosula, jujube and ginger on the spontaneous activity of gastric strips were not obvious, but they had obvious antagonistic effects on the spastic contraction of gastric strips caused by acetylcholine chloride. Both Evodia rutaecarpa and Evodia rutaecarpa Decoction showed a decrease in tension and contraction amplitude, and a decrease in frequency.

2.3.3. effects on the isolated small intestine activity caused by various drugs: rabbits are about 1.5kg and guinea pigs are about 250g. They are prepared and tested according to the method of warm blood animal ex vivo intestinal tube experiment, and the spontaneous contraction activity is recorded. After it is stable, the drugs to be antagonized (nicotine, physostigmine, acetylcholine, phentolamine, reserpine, sodium chloride and histamine) are given. After the recorded contraction activity is stable, Evodia rutaecarpa decoction is given once every 3 minutes for a total of 3 times (except that a single dose is used to antagonize acetylcholine and histamine), so that the nutrient solution presents three cumulative concentrations of traditional Chinese medicine. After each administration of traditional Chinese medicine, the changes in intestinal activity are recorded for 3 minutes. The results showed that Evodia rutaecarpa gradually inhibited the activity of small intestine in vitro induced by seven drugs with increasing dose.

2.3.4. antiemetic effect: 30 healthy domestic pigeons, either male or female, weighing 300-400g, were randomly divided into three groups, 10 in each group. The experimental group was given Evodia rutaecarpa and Evodia rutaecarpa Decoction respectively, and the control group was given tap water by gavage according to the same volume. One hour after administration, each pigeon was gavaged with 2% copper sulfate 1ml / 100g body weight. The time of the first vomiting (vomiting latency) and the number of vomiting (vomiting frequency) within 1 hour after the administration of copper sulfate were recorded for each pigeon. . Evodia rutaecarpa also has a certain antiemetic effect, but the statistics are not significant.

2.3.5. antidiarrheal effect:

2.3.5.1. effect on diarrhea in mice caused by castor oil thirty six male and female mice, weighing 20.7 ± 2.6g, were divided into three groups. They were respectively gavaged with Evodia rutaecarpa Decoction and tap water, and then enema with castor oil 0.15ml / mouse after 30 minutes, and the number of diarrhea was recorded. During the experiment, food and water were forbidden. .

. It can be seen that the anti diarrhea effect of Evodia rutaecarpa increases with the increase of dose, and the effect is slow, but lasts for a long time.

3. effects on late hemorrhagic shock: 22 large eared white rabbits, regardless of sex, weighing 1.75 ± 0.25kg, were used to replicate the late hemorrhagic shock rabbit model by imitating the Wiggers blood storage bottle method. After whole blood transfusion, the drug was started to be administered from ear marginal vein. The treatment group was given Wuzhuyu Decoction injection (normal saline diluted to 0.5g / ml) 10ml/kg; 4 drops / min, 3 hours; The control group was given the same amount of normal saline, and the 3-hour and 6-hour survival rates, mean arterial blood pressure, heart rate and urine output were observed. . The 3-hour survival rate in the treatment group was 11 / 11, which was significantly higher than 6 / 11 in the control group (P < 0.05); The 6h survival rate was 7 / 11, which was also significantly higher (P < 0.05) than 1 / 11 in the control group. . After administration, the breathing and heartbeat in the treatment group changed from weak to clear, and the limbs shook; The breathing and heart rate of rabbits in the control group were still weak, and their limbs were stiff.

4. effect on thrombosis and coagulation function: Sixty Wister rats, half male and half female, weighing 185 ± 25g, were randomly divided into six groups. The control group was given equal volume of normal saline; Evodia rutaecarpa water extract group was administrated by gavage at 1 / 5, 1 / 10, 10-20g/kg of LD50, and the other two groups were aconite water extract group. One hour after administration, the time of thrombosis was measured, and the blood was taken to determine the biochemical items. Heparin group, intravenous injection of 100u/kg, determined by the above method. Results the thrombosis time of Evodia rutaecarpa aqueous extract was significantly prolonged at the dose of 20g / kg; At the dose of 10G / kg, the partial thromboplastin time (kptt) of white clay was prolonged to varying degrees, and at the dose of 20g / kg, kptt and VF were significantly prolonged. .

5. hepatoprotective effect: effect on SGPT and SGOT in rats with CCl4 induced acute liver injury thirty six male and female rats weighing 222 ± 25g were equally divided into four groups. They were gavaged with Evodia rutaecarpa Decoction and tap water once a day for five consecutive days. One hour after the last administration, except the tap water control group, the other groups were intraperitoneally injected with carbon tetrachloride (CCl4) 0.3ml / kg, and after fasting for 16 hours, SGPT and SGOT were measured by carotid artery blood sampling under urethane anesthesia. It can be seen that Evodia rutaecarpa 5 and 10g / kg can resist the effect of CCl4 increasing SGPT, and the 5g / kg group can not resist the effect of CCl4 increasing SGOT.

6. other effects: intragastric administration of 20g/kg Evodia rutaecarpa Decoction to mice can significantly reduce the number of writhing reactions caused by potassium antimony tartrate and prolong the latency of pain response to hot plate stimulation. It has been reported that both methanol and water extracts of Evodia rutaecarpa have serotonin receptor affinity, indirectly suggesting that dehydroaevodiamine may be a serotonin receptor agonist. . The cytotoxin isolated from this plant is a yellow powder. In the cytotoxicity test with V-79 cells, only this yellow powder showed obvious activity. In further cytotoxicity tests with tumor cells, human nasopharyngeal carcinoma (KB), murine lymphocytes and leukemia cells (P388), only this yellow powder showed activity. Their IC50 values were: V-79 was 0.19mg / ml; KB was 0.98mg / ml; . In addition, evodiamine in Evodia rutaecarpa fruit has anti hypoxia effect.