Cornus officinalis
1、 Pharmacopoeia standard of Cornus officinalis . Harvest the fruit when the peel turns red in late autumn and early winter, bake it with slow fire or place it in boiling water for a little while, remove the core in time and dry it. [character] the product is irregular in shape of sheet or capsule, 1-1.5cm long and 0.5-lcm wide. The surface is purplish red to purplish black, wrinkled and shiny. Some have round persistent calyx marks at the top and fruit stem marks at the base. It is soft. It is slightly sour, astringent and bitter. [identification] (1) the powder of this product is reddish brown. . Mesocarp cells are orange brown and often wrinkled. Calcium oxalate cluster crystals are few, with a diameter of 12 ～ 32 μ M. Stone cells are square, oval or rectangular, with obvious pits and large cell cavities. (2) Take 0.5g of this product powder, add 10ml of ethyl acetate, sonicate for 15 minutes, filter, evaporate the filtrate, add 2ml of absolute ethanol to dissolve the residue as the test solution. In addition, take ursolic acid reference substance and add absolute ethanol to make a solution containing 1mg per 1ml as the reference substance solution. . In the chromatogram of the test sample, the same purple red spots appear at the corresponding positions of the chromatogram of the control sample; Put it under the ultraviolet light (365nm) to inspect, showing the same orange yellow fluorescent spots. (3) Take 0.5g of this product powder, add 10ml of methanol, sonicate for 20 minutes, filter, evaporate the filtrate, add 2ml of methanol to dissolve the residue as the test solution. In addition, take the morroniside reference substance and loganin reference substance, add methanol to make a mixed solution containing 2mg per 1ml each as the reference solution. According to the test of thin-layer chromatography (general rule 0502), absorb 2 μ l of each of the above two solutions, respectively dot on the same silica gel G thin-layer plate, use chloroform methanol (3:1) as the developing agent, develop, take out, dry, spray with 10% sulfuric acid ethanol solution, heat at 105 ℃ until the spots are clear, and check under the ultraviolet 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. [inspection] the impurities (stone and stem) shall not exceed 3% (general rule 2301). The moisture content shall not exceed 16.0% (the second method of general rule 0832). The total ash content shall not exceed 6.0% (general rule 2302). . [content determination] determine according to HPLC (general rule 0512). Chromatographic conditions and system suitability test octadecylsilane bonded silica gel was used as filler; Acetonitrile was used as mobile phase a, and 0.3% phosphoric acid solution was used as mobile phase B. gradient elution was performed according to the following table; The detection wavelength was 240nm, and the column temperature was 35 ℃. The number of theoretical plates shall not be less than 10000 according to loganin peak. ----------------------------------------. 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 80% methanol, weigh it, heat and reflux for 1 hour, cool it, then weigh it, make up the lost weight with 80% 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 products, the total content of monooside (c17h26o11) and loganin (c17h26o10) shall not be less than 1.2%. [processing] Cornus officinalis flesh to remove impurities and residual fruit stones. [character], [identification], [inspection] (moisture and total ash) [content determination] is the same as that of medicinal materials. . . It has a slight wine aroma. [content determination] the total amount of monooside (c17h26o11) and loganin (c17h26o10) in the same medicinal material shall not be less than 0.70%. [identification], [inspection] (moisture and total ash) [extract] is the same as that of medicinal materials. [nature, taste and meridian tropism] sour, astringent, mild. Return to liver and kidney channels. [function and indications] tonify the liver and kidney, astringent and firm. It is used for dizziness and tinnitus, waist and knee soreness, impotence and spermatorrhea, enuresis and frequent urination, metrorrhagia and metrorrhagia, sweating and collapse, internal heat and thirst. [usage and dosage] 6 ～ 12g. [storage] store in a dry place to prevent moths.

2、 Chemical constituents of Cornus officinalis
Fruit containsCornus officinalis glycoside(cornin i.eVerbenosideVerbe-nalin）、 Saponins, tanninsUrsolic acid（Ursolic acid）、gallic acid, malic acid, tartaric acid and vitamin A. the fatty oil of seeds containspalmitic acid, oleic acid and linoleic acid, etc. Fruit containsMorifolin（morroniside）、7-o-methylmorolonidin(7-O-methylmorroniside)、Angelica glycosides(sweroside) andChitin(loganin)。 In addition, containingUrsolic acid(ursolic acid)、 Tartaric acid, malic acidgallic acid(gallic acid)、 Vitamin A and about 13% saponin. Cornus tannins 1,2,3 were also reported. Recently, bicyclic iridoid glycosides were obtainedCornus officinalis neoglycoside.The new glycoside of Cornus officinalis L.in the new period of development of Cornus officinalis L.in the new period of development, the new glycoside of Cornus officinalis L.in the new(cornuside)。 Fruit stone containsβ - sitosterol、Betulinic acid、Ursolic acid、Methyl gallate、gallic acidEt al. Seed containsoleanolic acid (oleanolic acid) andUrsolic acid。

3、 Pharmacological effects of Cornus officinalis
1. antibacterial effect fruit decoction can inhibit the growth of Staphylococcus aureus in vitro, but has no effect on Escherichia coli.

2. hypoglycemic effect
  2.1 effect on blood glucose of normal rats male Wistar rats, weighing 150-200g, were divided into 4 groups equally. Group 1 was the control group, and the other three groups were the drug groups, with 5, 7.5, and 15g / (kg, d) of drug (Cornus officinalis mature pulp). After milling, 70% ethanol was used to reflux and extract, and 1ml of extract (equivalent to 3.5G of crude drug) was prepared. There was no significant difference in blood glucose between the drug group and the control group. In addition, 50g/kg drug was used to treat rats with Ig1 for one week, and the results also showed that it had no obvious effect on the blood glucose value of normal rats.

  2.2 effects on hyperglycemia induced by adrenaline in rats 24 rats were equally divided into three groups. After 2 hours of fasting, the normal control group and the modeling control group were given Ig water, and the administration group was given Ig Cornus 7g/kg. One hour after administration, rats in the modeling control group received SC normal saline; The SC adrenal glands of rats in the modeling control group and the administration group were 0.25mg/kg. Blood glucose was measured 135 minutes after injection. It can be seen that Cornus officinalis has obvious effect on anti adrenaline induced hyperglycemia.

  2.3 effects of alloxan on hyperglycemia, liver glycogen, triglycerides and cholesterol in rats 24 rats were equally divided into 3 groups. One group was the normal control group, the other two groups were treated with 30mg/kg of alloxan normal saline solution under the tongue of rats, and one group was treated with 7g/kg of Cornus officinalis daily. .

  2.4 effects on azosulfonamide induced diabetes in rats Wistar male rats were used. Diabetes mellitus was induced by 50-65mg/kgiv azosulfonamide after the animals stopped eating for 45 hours. Bawei Dihuang Pill was made into corresponding dry extract and powder, and other components were made into powder. . Then, three kinds of crystalline fish liver acid oil, ursolic acid and 2-a-hydroxyursolic acid were prepared from coe-fr-2. The first two were used for experiments. The powder of Cornus officinalis and Bawei Dihuang pills was 2g/kg, coe1g/kg, and the three subfractions of CoE and fish liver oleic acid and ursolic acid were 0.5g/kg, respectively. The method of administration was as follows: animals were Po administered 1h after intravenous injection of streptozotocin, and then twice a day for a total of 6 times. Blood was taken from the abdominal vein 3 hours after the last Administration for blood glucose measurement. At the same time, the urine sugar and 20-hour water intake and urine output of the animals were measured. The experimental results showed that the anti diabetic effect of Bawei Dihuang Pill powder and Cornus officinalis was the most obvious. After the administration of Cornus officinalis powder, the water consumption of animals in 20 hours was 23.3 ± 2.8, which was significantly different from that of the control group (46.4 ± 50) (P < 0.0001 =). The blood glucose (mg/dl) and urine glucose content (mg/20 h urine) were 393.3 ± 28.7 and 2553.0 ± 205.1, respectively, which were significantly different from those of the control group (470.0 ± 21.3 and 4733.7 ± 389.5, P < 0.05 for the former and P < 0.0001 for the latter). Bawei Dihuang Pill powder has no obvious hypoglycemic and urinary glucose effects, while its dry extract has no obvious effect. Among the ether extractants of Cornus officinalis, coe-fr-2 was the most active. After the administration of coe-fr-2, the 20-hour water intake and urine output of the animals were 13.8 ± 3.4 and 13.5 ± 1.9, respectively, which were significantly different from those of the control group (50.0 ± 3.2 and 36.7 ± 3.0) (P < 0.001 =), and the blood glucose and urine glucose contents were 227.2 ± 25.3 and 978.2 ± 167.8, respectively, which were significantly different from those of the control group (403.3 ± 36.6 and 2916.8 ± 55.4) (P < 0.05 for the former and P < 0.0001 for the latter). In addition, CoE and coe-fr-l also had obvious hypoglycemic effect, while CoE and coe-fr-2 had a tendency to reduce animal weight. For the fish liver acid oil and ursolic acid isolated from coe-fr-2, although both can significantly reduce the 20-hour water intake and urine output of animals, only ursolic acid can significantly reduce blood glucose and urine glucose. After administration of ursolic acid, the blood glucose and urine glucose contents of animals were 294.2 ± 37.2 and 1175.5 ± 249.2, respectively, which were significantly different from those of the control group (513.2 ± 35.4 and 3013.3 ± 404.2) (P < 0.0001). 2.5 effect of ursolic acid on whole blood viscosity in hyperglycemic rats after alloxan induced diabetic rats were Ig Cornus officinalis 7g/kg for 1 week, blood glucose was measured by tail vein blood sampling. Then anesthetized with 40mg/kg sodium pentobarbital solution, anticoagulated with heparin, blood was taken from abdominal aorta, and whole blood viscosity was measured.

2.6 effect on platelet aggregation in hyperglycemic rats Cornus officinalis 7h/kg, two hours after the last Ig alloxan induced diabetic rats, anticoagulation was performed with 3.8% sodium citrate at a ratio of 9:l, and blood was taken from the abdominal aorta to prepare platelet rich and platelet poor plasma. ADP was used as the aggregating agent to determine the effect of the drug on platelet aggregation function. Results the platelet aggregation inhibition rate of Cornus officinalis group was 44.6% (P < 0.00013). Effect on immune system 3.1 effect on the weight of immune organs forty male ICR mice were randomly divided into four groups. The experimental group received 5dig Cornus officinalis 10.20g/kg (water decoction) continuously; . Twenty four hours after the last administration, the animals were sacrificed by exsanguination, and the thymus, spleen and liver of mice were harvested and weighed to calculate the index of each organ. Cornus officinalis reduced the thymus index of mice, but had no significant effect on liver and spleen.

3.2 effect on clearance rate of carbon particles take male ICR mice. The dosage and mode of administration of Cornus officinalis and cyclophosphamide are the same as above. . Results the K value of different doses of Cornus officinalis group was significantly lower than that of the control group.

3.3 effects on mouse serum antibodies 3.3.1 effects on hemolysin antibodies male NIH mice were treated with 5.10g/kg of Cornus officinalis for 5 days in the administration group and 25mg/kg of cyclophosphamide for IP in the positive control group from the second day before sensitization with sheep red blood cells (SRBC). 24h after each administration, HC50 of samples was measured and calculated according to the colorimetric method. It can be seen from table 6 that 10g/kg of Cornus officinalis significantly increased the content of serum hemolysin antibody in mice, while 5g/kg of Cornus officinalis had no significant effect.

3.3.2 effect on the content of antibody IgG balb/c mice, both male and female, the dosage of Cornus officinalis is the same as above, and the positive control group started from the fourth day of administration of cyclophosphamide 100mg/kg for 2 consecutive days in the experimental group. Twenty four hours after the last administration, blood was collected from the mouse orbit, and the serum was separated. The IgG content was measured by agar unidirectional diffusion method. The IgG content was expressed by the diameter of the precipitation ring. Different doses of Cornus officinalis could significantly increase the serum IgG content of mice.

3.4 effect on delayed foot pad reaction induced by SRBC male Kunming mice were selected. The experimental group was given the drug 1D before SRBC sensitization. The dosage and method of Cornus officinalis were the same as that of 3. After the last administration, the mice were challenged with antigen 1H later. The thickness difference and weight difference of the left and right foot pads of mice were measured 24h later. Results Cornus officinalis at 5 and 10g/kg inhibited the delayed foot pad swelling induced by SRBC in mice.

3.5 effects on contact dermatitis caused by DNCB 3.5.1 effects of administration before challenge on contact dermatitis female NIH mice were selected. The experimental group started from D6 after DNCB sensitization, and continuously applied 5dig of Cornus for 10, 20d/kg. 6h after the last administration, DNCB was applied to the right ear of mice for attack. After 16h, two ears were cut off, ear pieces were punched with a 9mm diameter punch, and the degree of contact dermatitis was expressed by the weight gain of the right ear. Results different doses of Cornus showed obvious inhibitory effect on auricle swelling in mice.

3.5.2 effect of administration on contact dermatitis after challenge fifty six male NIH mice were randomly divided into seven groups. The experimental method was the same as the literature. At dl0 after DNCB sensitization, the administration group was Ig Cornus 20 g / kg at different times after antigen challenge. Cornus officinalis can significantly inhibit DNCB induced contact dermatitis in mice when administered 3-15 hours after challenge.

The total glycosides of Cornus officinalis had immunosuppressive effect on mice in vitro and in vivo; In vitro test, ursolic acid can kill cultured mouse lymphocytes and lose the ability of lymphocyte transformation, IL-2 production and LAK cell production. However, if ursolic acid is injected intraperitoneally, the above indicators of mice are significantly improved.

4. anti shock effect
4.1 effect on carotid blood pressure healthy rabbits of 1.5-3.5kg, either male or female, were randomly divided into two groups, group A as the experimental group and group B as the control group. Carotid artery dissection was performed under local anesthesia with 2% procaine, intubated, connected to rm-600 four channel physiological recorder, and 312u/ (M1, kg) heparin was injected into ear vein. Then the femoral artery was slowly bled to reduce the blood pressure to less than 50mmhg. Due to the compensatory function of rabbits, blood pressure can rise automatically after stopping bloodletting. If the recovery was high, the blood was bled again to maintain the blood pressure below 60mmhg for 1h, and the values of various observation indicators during the shock period were recorded. Then group A was given 1g/ml Cornus injection through the subcutaneous superficial vein of the neck, and the administration speed was 36 drops / min; Group B was given the same amount of normal saline intravenous drip. . The specimens were taken after 30 minutes, and the observation indexes were recorded after treatment, and the observation was terminated. The results showed that in group A, the average value of 49.73 ± 7.25mmhg increased from shock stage to 87.64 ± 7.13mmhg, with an increase of 39.43 ± 9.97mmhg; In group B, the average increase from 55.46 ± 8.57mmhg in shock phase to 68.61 ± 17.6mmhg, with an increase of 13.15 ± 12.12mmhg. There were significant differences between the two groups, t=61.8,P＜0.001。

4.2 effect on the amplitude of blood pressure heartbeat wave in the above experiment, group a increased from an average of 1.42 ± 0.7mm in shock phase to 4.03 ± 1.11mm, with an increase of 2.61 ± 0.72mm; group B increased from an average of 1.88 ± 0.9mm in body weight phase to 2.35 ± 0.87mm, with an increase of 0.57 ± 0.93mm. There was significant difference between the two groups, t=5.36，P＜0.001。

5. inhibition of inflammatory response
5.1 inhibition of peritoneal capillary permeability in mice male Kunming mice, weighing 18-22g. The positive control group received 50mg/kg of Ig aspirin daily; In the administration group, Cornus officinalis was Ig 5, 10g/kg (the decoction contained crude drug 1g/ml) daily for 5 days; . It can be seen that different doses of Cornus officinalis can significantly inhibit the increase of peritoneal capillary permeability induced by acetic acid in rats.

5.2 inhibition of xylene induced ear swelling in mice male Kunming mice, weighing 18-22g, were treated with 5dig of Cornus officinalis 10.20g/kg continuously. One hour after the last administration, 30ul of xylene was applied to the right ear of mice, and the left ear was used as a control. Two hours later, the ear was cut off and the ear piece was removed with a 9mm diameter puncher and weighed. Results the weight difference (mg) of left and right auricles in the control group and Cornus 10.20g/kg group were 18.2 ± 4.5, 9.9 ± 4.5 (P < 0.01), 11.3 ± 4.3 (P < 0.01), (n = 10), respectively. It showed that Cornus officinalis significantly inhibited xylene induced ear swelling in mice.

5.3 inhibition of inflammasome induced rat footpad swelling male SD rats, weighing 120-160g, in the positive control group and the administration group, 50mg/kg of aspirin and 5g/kg of Cornus officinalis were administered continuously. One hour after the last administration, 0.1ml of fresh egg white was prepared from SCL% carrageenan on the rat right rear footpad. The degree of footpad swelling (mm) was measured every hour after inflammation. Results Cornus officinalis 5g/kg had a significant inhibitory effect on the swelling caused by egg white, while the inhibition effect of carrageenan induced footpad swelling was only significant at 1 hour.

5.4 inhibit the proliferation of rat cotton ball granulation tissue male Wistar rats, weighing 150-220g, were anesthetized with ether, and 20mg of heavily disinfected cotton balls were embedded under the armpits of both forelimbs. The positive control group and the administration group were continuously SC hydrocortisone 10mg/kg and Ig Cornus 5, 10g/kg, respectively. The rats were sacrificed on D8, and the granulation tissue was stripped. After baking at 80 ℃ for 3 hours, they were weighed, and at the same time, the thymus and adrenal glands of each rat were weighed. It can be seen from table 12 that Cornus officinalis significantly inhibited the proliferation of cotton ball granulation tissue in rats, and had no significant effect on the weight of thymus and adrenal gland.

5.5 reduce the content of ascorbic acid in rat adrenal gland take the right adrenal gland of rats used for granuloma test, make homogenate, and determine the content of ascorbic acid according to the 2,4-dinitrophenylhydrazine color development method. .

5.6 effect on PGE content in inflammatory tissues of rats male SD rats, weighing 150-180g, were given 5dig of Cornus officinalis 10g/kg or indomethacin 50mg/kg consecutively in the administration group and the positive control group respectively. 30 minutes after the last administration, 0.1ml of SCL% carrageenan was placed on the right hind foot of each rat. After 3 hours, they were sacrificed. The right hind foot was cut off from the LCM above the ankle joint and weighed. After peeling, it was placed in 5ml of normal saline for 1H. 0.15ml of the supernatant was added to 1ml of 0.5mol/l potassium hydroxide methanol solution. It was isomerized in a water bath at 50 ℃ for 20min, and methanol was 2.5ml, and the UV absorption value (△ a) was measured at 278nm. The PGE content was expressed by the equivalent absorption value per gram of inflammatory tissue. Results dogwood 10g/kg had no obvious effect on the weight of inflamed feet and PGE content in rats.

6. anticancer effect inoculate healthy mice with 0.1ml of ascites cancer cell fluid. After 7-8 days of growth, there is more cancer fluid. Draw out the cancer fluid and add a drop to the glass slide, and add a drop of 20% Cornus Decoction and a drop of eosin tais solution. Observe the death of cancer cells under the microscope. If killed, the cells will be stained red, and if not dead, they will still be colorless and transparent. The normal cells of the control were salivary gland cells and testis cells. The experimental results showed that Cornus officinalis could kill all cancer cells; Testis cells also have the same effect, but only a small part of salivary gland cells are killed.

. The above acidic solution has little toxicity to frogs, mice, rats and rabbits, has no effect on body weight and blood picture, and can cause mild congestion of rabbit gastric mucosa, but has no effect on rabbit conjunctiva. Cornus officinalis glycosides have low toxicity and no hemolysis, but have weak effect of exciting parasympathetic nerves. This product has antioxidant effect.