2. Stir-fry the earthworm segments with yellow wine until they are slightly stuffy, put them in a pot and fry them with slow fire until they are slightly dry, take them out and let them cool (yellow wine for earthworm 100kg). 3. Stir-fry with wine and sand, mix well with wine, and get slightly bored. Put the sand in a hot pot, stir-fry until it is smooth, add Sheng Dilong, stir-fry until the surface is brown and slightly swollen, take it out, sieve out the sand, and let it cool. 4. Honey bran frying method: evenly spray the earthworm segments with wine (100: 15) and moisten1h.. Then put the honey bran into the pot and fry until it starts to smoke slightly, that is, put it into the earthworm and stir it constantly. When the surface of the earthworm is brown, take it out, quickly pour it into a container, cover it with a sack and stew it for 5 ~ 65438+. At the same time, we tried to cook in an electric incubator, mixing earthworm with old rice vinegar (1∶0.2) and wetting1h. After the vinegar is completely absorbed, spread it in an enamel dish with a thickness of about 3cm, put it in an electric thermostat at 100℃ for 2 hours, turn it once in the middle, bake it until the surface of the earthworm is brown, take it out and spread it out. Dry coarse powder of body cavity fluid of earthworm and ginseng was directly injected into the duodenum of rats at a dose of 30mg/kg. Fibrinogenesis was determined by the method of connecting fibrin in series with arteriovenous cannula, which showed obvious fibrinolytic activity, and the activity was lost after boiling or drying. The fibrinolytic activity of 5μg freeze-dried coarse powder is equivalent to that of urokinase1u. However, after the fibrin plate was heated in 85℃ water bath for 30 minutes, the activity of urokinase was lost, while the activity of freeze-dried coarse powder was partially retained. It shows that the fibrinolytic mechanism of freeze-dried coarse powder is not exactly the same as that of urokinase. The function of urokinase is to activate plasminogen into active fibrinolytic solution and catalyze fibrinolysis. In addition to urokinase-like fibrinolysis, earthworm freeze-dried coarse powder also directly promotes chemical fiber dissolution. Eisenia foetida extract can dissolve rabbit venous thrombosis in vitro, and also has fibrinolytic activity to coagulated fibrin glue plate. Its fibrinolysis has indirect effects of direct fibrinolysis and activation of plasminogen, but direct fibrinolysis is the main one. 0.lg/kg or 0.2g/kg of earthworm water extract was injected intravenously into rabbits, which obviously inhibited the experimental thrombosis of common carotid artery-external jugular vein blood flow, and also significantly inhibited 1g/kg, but it was weak. Intravenous injection of 0.02 g/kg has thrombolytic effect. Intravenous injection of 2 ml/kg earthworm ethanol extract injection (containing 2 g crude drug per 65438±0ml) in rabbits can obviously prolong the time of platelet thrombosis and fibrin thrombosis, reduce the length and dry weight of thrombus and reduce blood viscosity. Intravenous injection of artificially cultured crude extract of earthworm feces can significantly reduce the content of fibrinogen in blood and shorten the dissolution time of globulin.
After further extraction, the preparation containing various plasmin and plasminogen activator has good thrombolytic effect. It can improve the activity of tissue plasminogen activator (t-PA) in rabbit plasma, significantly reduce platelet aggregation, whole blood viscosity and plasma viscosity, and reduce the rigidity index of red blood cells, suggesting that it has the functions of promoting fibrinolysis, inhibiting platelet aggregation and enhancing the stability of red blood cell membrane. The water extract of earthworm dry powder can dissolve rabbit fibrin and blood clot in vitro. Intravenous injection of water decoction and alcohol precipitation in rabbits has obvious dissolving effect on venous thrombosis of ear margin. The earthworm extract taken orally by normal volunteers also has obvious anticoagulant and fibrinolytic effects in a dose-dependent manner. The time of thromboplastin was obviously prolonged, the activity of plasminogen activator was enhanced, the dissolution time of globulin was shortened, and the content of fibrin degradation products was increased. Fibrinolytic enzyme (freeze-dried powder) extracted from earthworm was gavaged to rabbits at 0.05 g/(kg d) or 0.1g/(kg d) for 3 days. Taking blood from the heart, the length, wet weight and dry weight of thrombus formed in vitro were significantly reduced compared with the control group.
Tommy Tam decoction has obvious antithrombin effect on bovine thrombin promoting coagulation of human fibrinogen in vitro. Eisenia foetida extract can obviously prolong thrombin time (TT), and it can also obviously prolong TT when added to plasma lacking antithrombin -III(AT-III), but the extension degree is small, which indicates that this effect has two parts: AT-III dependence and AT-III independence. Intravenous injection of earthworm extract in rabbits can obviously prolong TT half-life by 0.32 hours, and in vitro injection of earthworm extract can also obviously prolong TT in white mud and activate partial thromboplastin time (KPTT). The aqueous extract of Pheretima was hydrolyzed by alkali, and then separated by ethanol. The obtained extract can also significantly prolong KPTT and TT, and its antithrombin activity is not neutralized by AT-III antibody and protamine, indicating that its activity is independent of AT-III. The active ingredient is not polysaccharide or protein, but may be a small peptide with alkali resistance and heat resistance or a compound with double bonds, which is different from anticoagulant substances such as bombesin and heparin. Patients with hyperviscosity and ischemic stroke were treated with Eisenia foetida extract 14d for three courses and1~ for two courses, and their blood cell ratio, erythrocyte sedimentation rate, plasma viscosity and fibrinogen decreased. Intravenous injection of earthworm extract in rabbits can significantly reduce platelet aggregation rate, whole blood viscosity, plasma viscosity and erythrocyte rigidity index. (1) has antiarrhythmic effect on animals.
Intravenous injection of earthworm has obvious antagonistic effect on arrhythmia induced by chloroform-epinephrine or aconitine in rats and arrhythmia induced by barium chloride or poison hairy flower G in rabbits. It also inhibits cardiac conduction.
(2) Hypotensive effect: Anesthetized dogs were injected with 0. 1g/kg hot water extract or ethanol extract intravenously, and their blood pressure decreased within 30-45 minutes, lasting for 2-3 hours; Normal rats were given high dose (10g/kg) three times or low dose (50mg/kg) twice a day, all of which had obvious antihypertensive effect. The blood pressure of renovascular hypertensive rats decreased from 3 to 7 days after drug administration, and rose to the level before drug administration in the second week after drug withdrawal. The hypotensive mechanism of Pheretima may be that it directly acts on the central nervous system above the spinal cord or affects the central nervous system through some receptors, causing some splanchnic blood vessels to dilate and lower blood pressure. Oral administration of 0.25g/kg of Guangdilong decoction in rats immediately caused a decrease in blood pressure, which was similar to that of platelet activating factor (PAF). If CV6209, a PAF receptor blocker, is injected intravenously in advance, the antihypertensive effect of Pheretima angustifolia is significantly inhibited, suggesting that PAF-like substances are important antihypertensive components of Pheretima angustifolia. Feeding earthworm-containing feed to stroke-prone spontaneously hypertensive rats (SHRSP) can reduce the blood pressure of SHRSP and improve vascular elasticity, that is, increase the wet-to-weight ratio of aortic load (Raqq/wet-weight mg of aorta) and the maximum aortic load (Raqq/wet-weight mg of aorta) at rupture. Mice were given earthworm extract l 1mg/ mouse every day for 58 days, and treated with transplanted sarcoma S 180. It has obvious inhibitory effect, which not only makes the tumor shrink obviously, but also prolongs the survival time of mice. Feed earthworm extract to mice every day.
1mg/ animal for 6 days for transplanted solid tumor H22. It also has obvious inhibitory effect. Inoculate the dialysate of earthworm extract after dialysis with sarcoma S 180 or solid tumor EMT6. All mice have the effect of inhibiting tumor growth, and the anti-cancer effect disappears after 100℃ heating for 20min minutes. The components of earthworm dialysis can obviously inhibit thymidine (H3) infiltration in gastric cancer cell MGC803 in vitro. The component still has anti-tumor effect after heating at 56℃ for 0.5h, which shows that the effective component has small molecules and heat resistance. The earthworm extract was separated by gel filtration technology and four components were obtained. Component I has a very significant inhibitory effect on 3H-TDR infiltration of gastric cancer cell MGC803 in vitro. After heating at 56℃ for 0.5h, the anti-tumor effect of the component still exists, but it is obviously weaker than that of the unheated group, suggesting that the anti-tumor effect of the component remains partially after heating and has certain heat resistance. Component IV can also significantly inhibit the 3h-TDR infiltration of gastric cancer cell MGC803 in vitro, but after heating at 56℃ for 0.5h, the anti-tumor effect disappears, suggesting that this component is not heat-resistant, while components II and III have no anti-tumor effect. Earthworm extract has a concentration-dependent inhibitory effect on laryngeal cancer cell Hep-2 in vitro. The combination of earthworm extract and 5- fluorouracil (5-Fu) has obvious synergistic effect on the treatment of breast cancer in mice. The growth of tumor was delayed by 3 days compared with 5-Fu alone, and the cancer cells degenerated and died.
The single use is serious. For mouse sarcoma S 180, the mice can survive for a long time by using earthworm extract and interleukin -2 (IL-2) alone. It can also inhibit tumor growth and show synergistic effect. Transplant sarcoma S 180. When 5-Fu is combined with earthworm extract, it has synergistic effect on tumor inhibition. If it is combined with radiation synergist AK 123, the radiation synergistic effect is stronger, and the combination of the three has the best antitumor effect. In vitro, earthworm extract combined with AK 123 also has synergistic effect on human laryngeal cancer cell Hep-2. The combination of earthworm extract and hyperthermia also has obvious synergistic effect on the inhibition of solid tumor EMT6 in mice.
Oral earthworm extract can promote the transformation of spleen lymphocytes in normal and tumor-bearing mice (S 180), and can also promote the transformation of spleen lymphocytes in mice irradiated by linear electron accelerator and unirradiated mice. However, in vitro experiments can inhibit the transformation of mouse spleen lymphocytes and human blood lymphocytes. It is suggested that earthworm extract may inhibit the growth of tumor cells by improving the immune ability of the body. Intraperitoneal injection of earthworm extract (100 mg/kg) for 6 days can significantly increase the number of nucleated cells in thymus and spleen, and the reactivity of spleen cells to concanavalin A(ConA) and lipopolysaccharide (LPS) is significantly higher than that of the control group. Natural killer cells (NK) and antibodies in spleen are mediated by cells.
The cytotoxicity (ADCC) activity was also significantly higher than that of the control group. It has a similar effect on the immune function of tumor-bearing mice, but the activities of NK and AfXIC in spleen are significantly lower than those in control group. After local irradiation with deep X-ray therapy machine, the spleen weight, the number of nucleated cells in the spleen, NK cell activity, ADCC activity and brand cell IL-2 activity of tumor-bearing mice were all inhibited to some extent, but if treated with Dillon extract, there was no immunosuppression or even improvement, suggesting that it could promote the immune function of the body to some extent. Taking earthworm extract daily in mice can enhance the chemiluminescence of phagocytes in tail blood, suggesting that phagocytosis can be enhanced. The NK cell activity of tumor-bearing mice (S 180) decreased. If earthworm extract was given for 8 days, the activity of NK cells basically recovered. After taking the earthworm extract for 2 months, the tumor patients were tested for T3, T4, T8, T4/T8, Tb, Tac and B cell subsets by indirect immunofluorescence. Taking earthworm extract can increase T3, T4 and T4/T8, suggesting that earthworm extract is an immune enhancer. 150mg earthworm extract extracted from the eyes of patients with esophageal cancer who received radiotherapy twice a day can control the leukopenia of patients, and the effect is better than that of immune enhancement drug polyclonal antibody A.
After taking100 mg/kg for 7 days, the content of fluorescent products (directly reflecting the lipid peroxidation rate in vivo) increased due to the tumor-bearing decline, and glutathione reductase (GR), catalase (CAT), glutathione peroxidase (GSH-Px) and superoxide in blood moved.
The activity of superoxide dismutase (SOD) increased, suggesting that earthworm extract has the effects of scavenging free radicals and resisting lipid peroxidation. However, in vitro experiments show that earthworm extract, laser and blood leaf spray derivatives have synergistic killing effect on S 180 cancer cells, and the chemiluminescence intensity is also obviously enhanced. If free radical scavengers CAT and SOD were added to the culture medium, the enhanced luminescence intensity was obviously reduced, indicating that the synergistic effect of earthworm extract was at least partially realized by increasing active oxygen. Guinea pigs were intraperitoneally injected with earthworm solution (containing fresh earthworm about/kloc-0 per 65438 0 ml).
0.75g/ml)2ml, the incubation period of allergic asthma caused by ovalbumin was obviously prolonged, and the incubation period of asthma caused by histamine inhalation was also obviously prolonged, but it had no effect on asthma caused by choline acetate inhalation. Rats injected with 2ml earthworm solution intraperitoneally for 3 days can inhibit the passive skin allergic reaction caused by egg protein. In vitro, the earthworm solution with the concentration of 8× 10-7g/ml obviously inhibited histamine-induced tracheal contraction in isolated guinea pigs. The nitrogen-containing components extracted from earthworm have obvious bronchodilatory effect on rats and rabbits by lung perfusion, and can resist bronchoconstriction caused by histamine and pilocarpine. Through intravenous injection in guinea pigs, 50% animals can tolerate lethal dose of histamine. It is considered that a component in earthworm can block histamine receptor, antagonize tracheal spasm caused by histamine and increase capillary permeability, which is the main mechanism of antiasthmatic. Earthworm 3% acetic acid extract, 2.5% sulfuric acid extract.
84% ethanol and petroleum ether extracts have a strong inhibitory effect on human tuberculosis in vitro. The minimum dilution digits of the two substances extracted by 3% acetic acid to human tuberculosis are 100 letters and 70 times respectively, and the maximum dilution multiples of 2.5% sulfuric acid extract are 100 times, which are heat-resistant. The highest dilution ratio of 84% ethanol and pentaoleoether extracts is 3200 times, but they are not heat-resistant.
Lumbricus extract has a rapid killing effect on mouse and human sperm in vitro, among which ecdysterone acid and hyaluronic acid can quickly brake and agglutinate sperm and destroy its structure.
Earthworm extract is effective for local scleroderma. Biochemical analysis shows that the extract contains collagenase which specifically degrades collagen fibers, and its curative effect is probably caused by the degradation of locally denatured collagen fibers.