CN103239706B - Earthworm protein for reducing blood glucose, improving microcirculation and eliminating diabetic complication and application thereof - Google Patents
Earthworm protein for reducing blood glucose, improving microcirculation and eliminating diabetic complication and application thereof Download PDFInfo
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Abstract
The invention discloses an earthworm protein for reducing blood glucose, improving microcirculation and eliminating diabetic complication. The earthworm protein is prepared by the following steps of: 1) cleaning earthworms, namely selecting live earthworms, removing impurities and dead earthworms, putting the retained earthworms into water with the pH value of 4-5.8 for 10-15 minutes, adding 50 g of edible salt at the same time, then eluting the cleaned earthworms, flushing the earthworms by using warm water with the temperature of 28 DEG C so as to flush out residual water and impurities on the outer surfaces; 2) making pulp; 3) filtering homogenate obtained in the step 2) or performing centrifugal removal to remove rough slag in the homogenate, filtering the homogenate liquid subjected to slag removal to obtain a clean liquid, wherein every milliliter of clean liquid is more than 180 activity units, putting the clean liquid into a disinfection container, injecting high-pressure CO2 gas into the disinfection container, performing mechanical full stirring on the clean liquid to be treated for 3-8 minutes, outputting the clean liquid through a guide pipe to release pressure, wherein after the pressure is reduced, the CO2 solubility is quickly reduced, then filtering and cleaning the impurities, and precipitating the clean liquid for 12 hours to obtain supernate; and 4) freezing and drying the supernate, namely drying the supernate in a freezing dryer at the temperature of -28 DEG C to obtain powdered products, wherein every millimeter of the powdered products is equal to 2,000-3,000 activity units.
Description
Technical field
The invention belongs to field of health care products, relate in particular to a kind of blood sugar lowering, improve microcirculation, eliminate earthworm protein and the application thereof of complication.
Background technology
Earthworm protein is extracted from Pheretima, contains the Multiple components such as collagenase, fibrinolysin, Lumbrukinase, nucleic acid, trace element, and its molecular weight, at 5000-10000, belongs to short chain small-molecule substance.Pheretima extract (comprising earthworm protein etc.) has wide influence to the blood coagulation system in body and fibrinolytic system, both can obviously reduce rat platelet adherence rate, in extension body, thrombosis and dissolving thrombus in vivo, can increase again rat brain blood flow, reduces cerebral vascular resistance.Pheretima extract can also improve mouse red blood cell deformability, thereby improves hemorheological property and microcirculation disturbance.
Nutritive value
Rich in proteins in Pheretima body, in stem body multiselect 56～66%, is the half as much again of Semen sojae atricolor, Oryza glutinosa six times.In earthworm protein, contain multiple free amino acid, they are the topmost nutritional labelings of Pheretima.Its protein is made up of 18 seed amino acids, wherein the highest with the content of leucine and glutamic acid, every hectogram sample is respectively containing 22.161 and 6.227 grams, and next has aspartic acid, valine, lysine, arginine, alanine, glycine, methionine, serine, Soviet Union's chloric acid, phenylalanine, proline, histidine and tyrosine etc.Eight kinds of free amino acids of needed by human are comprised.Therefore, Lumbricus is a kind of full-valence animal egg Huang Yuan.The lipid part of Pheretima is containing the fatty acid of stearic acid, Palmic acid, highly unsaturated fatty acid, straight-chain odd number carbon and paniculate fatty acid, phospholipid, cholesterol etc.Highly unsaturated fatty acid is as higher in oleic acid, linoleic acid, arachidonic acid, linolenic content, has higher pharmacologically active, can prevent and treat atherosclerosis, is one of effective ingredient of Pheretima blood circulation promoting and blood stasis dispelling, control cardiovascular and cerebrovascular disease.Meanwhile, these unsaturated fatty acids also have certain Nutrition to human body.
Pheretima also contains the nucleotide compositions such as more hypoxanthine, xanthine, guanine, uracil, can provide requisite raw material for human body rib acid metabolic.Wherein. hypoxanthine or Pheretima blood pressure lowering, the bent active relative of relievining asthma are divided.In Pheretima, trouble has abundant fibrinolysin and catalase, peroxidase, dismutase, 8-D glucuronidase, alkali phosphatase. lipase, porphyrin synzyme etc.These enzymes are useful to human body.In Pheretima, contain abundant trace element as strontium, selenium, magnesium, zinc, copper, molybdenum, nickel, cobalt etc., these elements are all to ensure that health institute is requisite.
In earthworm protein, there is collagenase, fibrinolysin, earthworm protein, nucleic acid, the Multiple components such as trace element, its molecular weight is at 5000-10000, belong to short chain small-molecule substance, can enter smoothly blood capillary, dissolve the micro-bolt in blood capillary, allow the blood capillary of sclerosis recover elasticity, thereby remove microvascular obstruction, torsional deformation situation, directly dissolve the main substance fibrin that forms human thrombus, thereby the direct thrombolytic effect that performance is powerful, dissolve fresh and old thrombosis, can improve the anticoagulant function of human body self, can activate the differentiation characteristic of vascular endothelial cell, get through passages through which vital energy circulates, promote blood capillary regeneration, set up collateral circulation, for the heart and brain tissues of focal zone is opened up new blood access, repair downright bad cranial nerve cell, cure hemiplegia, thereby reach the effect that continues thrombolytic, to the prevention of cardiovascular and cerebrovascular disease, the rehabilitation of apoplexy sequela is significant.Contribute to again to eliminate the aging relevant with microcirculation disturbance and tumor generation, also improved whole body various physiological-function and health status, improve immunity simultaneously.
Thrombus
Earthworm protein fibre rich protein resolvase, can activate the fibrinoclase unit in thrombosis, makes it to be converted into the activated fibrinoclase of tool, and then fibrin in thrombus, fresh thromboembolism.Fibrinoclase in this product direct thrombus again, the thromboembolism of old.Thrombus does not affect the clotting mechanism in body, the side effect not causing bleeding.
Vessel softening
Active contained fibrinoclase and other active matter mass-energy are dissolved fibrin net in atherosis blood vessel wall, eliminate the pathological products such as attached cholesterol speckle thereon, promote synthesizing of vascular endothelial cell DNA, promote the reparation of vascular damaged, blood vessel dilating, recover blood vessel elasticity, make unblocked blood flowing, change to a certain extent and stoped arteriosclerotic natural history.
Reduce blood sticky
Take after active earthworm protein, along with the dissolving of thrombosis, softening, the microcirculatory improvement of blood vessel, reduce blood viscosity, make abnormal hemorheology index recover normal, Microcirculation of Nailfold is detected by extremely transferring to normal, thereby eliminated the hidden danger that cordis and cerebral accident occurs.
Two-ways regulation blood pressure
Active earthworm protein by recovering blood elasticity, blood vessel dilating, thrombus, improve microcirculation and improve circumference of cannon bone state, reduce blood flow Peripheral resistance, reduce blood viscosity and there is stable hypotensive effect.After essential hypertension is taken, can make controlling of blood pressure approach normal level after normal; Severe hypertension disease or cardiovascular and cerebrovascular vessel Complicated with Hypertension, take the hypotensive effect that can strengthen other depressor after this product, reduces its dosage and malicious 1 side effect, and this product is by improving blood quality and Peripheral resistance has regulating action to hypotension simultaneously.
Reduce blood glucose
The positive source comprehensive effect optimum due to active earthworm protein improved the multiple function of human body, improved the interior environment of body, promoted the recovery of islet function, has certain blood sugar reducing function after taking this product.
Property of raw material
Earthworm protein is extracted from Pheretima, and Pheretima has another name called Lumbricus, and [3] Pheretima is one of important Chinese crude drug of China.In 67 kinds of animal drugs recording in Chinese materia medica monograph Shennong's Herbal the earliest, just there is Lumbricus.Classify low-grades as at Shennong's Herbal, there is heat clearing away arresting convulsion, dredging collateral, relieving asthma, effect of diuresis.After normal parch for diseases such as high heat, coma, infantile convulsion tic, arthralgia, dyspnea and cough due to lung-heat, oliguria edema, hypertension.Main containing several amino acids.Because raw product fishy smell is too heavy, generally need be through parch therefore be used as medicine.TAO Hong-Jing meaning: " if take dry earthworm, must endure and consider to be worth doing ".Tradition is with Pheretima, and method for making is a lot, mainly contains medicine system, vinegar system, boils, the method such as processed with wine, oil system, Pulvis concha Mactrae parch, processed with salt, makes its quality pine steep crisp, detoxification, correct stink and be convenient to fried taking.
Pheretima is except having better nutritivity value, and it also has a lot of health cares, can strengthen the premunition of human body, improves people's health level.In recent ten years, Pheretima is carried out to large quantity research chemical, pharmacology aspect both at home and abroad, find that Pheretima not only contains abundant nutritional labeling, also contain multiple pharmacological component, there is many-sided pharmacological action, relate to each system of human body as cardiovascular system, blood system, immune system, central nervous system, respiratory system, digestive system etc.Especially unify aspect blood system at cardiovascular system, Pheretima is prophylaxis of hypertension, myocardial infarction, arrhythmia, apoplexy thromboembolism well.Application
Thrombus disease aspect
Earthworm protein has thrombolytic effect, and its effective ingredient is white enzyme of dawn.This kind of enzyme energy direct hydrolysis fiber dawn is white, and have fibre fight proenzyme invite live effect.Real fine horse result shows that the effective ingredient of this fibrinolysin thrombolytic is present in the 5th dawn white peak of the fine plain column chromatography of DEAE dimension and the 2nd dawn white peak of sepbadeG-50 column chromatography sympathized.This dimension lyase energy anticoagulant, anticoagulant, reduction blood viscosity, thus thrombosis stoped.
Cardiovascular and cerebrovascular disease aspect
Experiment shows, earthworm protein can obviously increase blood flow rate and the blood flow of rat serum circulation, reduce hematocrit percentage rate and the blood viscosity of cell, obviously improve percentage composition and the erythrocyte Mg of blood total protein ++ content, explanatorily some chemical composition of dragon protein really can be to some common cardiac and cerebral vascular diseases, as hypertension, coronary heart disease, arteriosclerosis and the fatigue disease of other blood etc., all there is good prevention effect.
Anti-tumor aspect
During by Pheretima extracting solution treatment gastric cancer, find, this extracting solution has significant inhibitory action to cancerous cell DNA synthetic; To mice with tumor treatment, after 38 days, result tumor cell disappears completely.Also have over the years many about with earthworm protein treatment kinds cancer patient and all obtain the clinical report of certain curative effect.This all explanatorily some contained chemical composition of dragon protein cancerous protuberance cell is had to very strong Inhibited differentiation and killing action.Or may remove the human blood hypercoagulability that cancer patient causes because of the solidifying factor of cancer cells secrete cancer due to some activation factor, thereby the therapeutic purposes that play blood circulation promoting and blood stasis dispelling, remove swollen dispersing tumor.
Earthworm protein has been proved to be good clinical value, and it is a kind of polymolecular restructuring oral formulations, has special affinity with thrombosis (fibrin), can follow the tracks of thrombolytic, effectively dissolve micro-bolt, improve microcirculation, add heart tonifying, cerebrovascular side Zhi Xunhuan, the open vascular injury endotheliocyte of repairing, increase blood vessel elasticity, improve blood vessel oxygen supply function, reduce blood viscosity, reduce platelet aggregation rate, suppress thrombosis and again form.Repair the downright bad brain cell of thrombosis generation rear perimeter edge, save half dark space.Be widely used at present clinically, and be used in more and more the heart, cerebrovascular, endocrine, the prevention and treatment of diseases centers such as respiratory system.
Current clinical trial shows, earthworm protein is definite to commonly encountered diseases therapeutical effect such as angina pectoris, cerebral infarction, diabetes, the nephrotic syndrome, pulmonary heart disease and deep vein thrombosis of lower extremity, and bad reflection is little, and patient easily tolerates.Although there still have many problems to need to be extensive, clinical control is researched and solved for a long time, looks forward to the future, and the application of earthworm protein in clinical disease has better prospect.
At present, the application of China's Chinese medicine has more and more obtained the accreditation of international community, its multiple medical material is evident in efficacy, medicine for blood sugar lowering, blood pressure lowering, blood fat reducing is various especially, but real evident in efficacy in practical application, only have several, such as Herb Gynostemmae Pentaphylli, Semen Cassiae etc., and the application mentions obtaining pulse family Cassia sophera Cassia sophera L., apply little.The applicant finds in the leaching process in earthworm protein in actual production and application, compound a certain amount of blood-sugar lowering tcm drug, the earthworm protein of locating of compound extraction has remarkable result aspect blood sugar lowering, in fat-reducing, blood fat reducing and blood pressure lowering, there is too significant curative effect, especially add after the batching of certain proportional quantity, successful is strengthened.
Earthworm protein is called as the king of thrombolytic in medical circle, be extracted from Lumbricus.Be applied to rapid osmotic behind affected part, thrombus becomes silted up and hinders, and makes unobstructed blood vessel, and varicosis falls after rise gradually.The aspects such as earthworm protein treatment diabetes, kidney syndrome have remarkable result equally.
Summary of the invention
For realizing object of the present invention, concrete scheme is as follows:
Blood sugar lowering, blood fat reducing, blood pressure lowering, improve microcirculation, eliminate the earthworm protein of diabetic complication, it is characterized in that being prepared by following steps:
1) Lumbricus is cleaned: select Lumbricus alive, remove impurity and the rotten Lumbricus of dead, it is 4～5.9 water 10～15 minutes that Lumbricus is put into pH value, adds 50g Sal simultaneously, the 2-3 hour of illumination simultaneously, impels Lumbricus get rid of the interior dirt of body and clean Lumbricus body surface; Lumbricus alive after cleaning is carried out to drip washing, use the warm water washing of 28 DEG C, wash away excessive moisture and the impurity of outer surface; 2) slurrying: by step 1) Lumbricus that obtains is positioned in container after weighing, meter is put into 30～50 parts by weight, pH value is 5.0～5.3 10～20 parts of sodium citrates, puts into 10～30 parts of Chinese medicine composite powders again, and described Chinese medicine composite powder is: 5 parts of 10 parts of Cassia sophera Linn seeds, 3 parts of Semen Vaccariae, 3 parts of the Radixs Astragali, 2 parts of Rhizoma Dioscoreaes, 2 parts, the Radix Rehmanniae, 2 parts of Fructus Schisandrae Chinensis, 1 part of Radix Ophiopogonis, 1 part of Fructus Mume, 1 part of Rhizoma Polygonati, 1.5 parts of Fructus Lyciis, 1 part of Radix Puerariae, 1 part, Folium Mori, Abelmoschus esculentus are crushed to 350～500 orders through super micron mill and make powder and stir and be composited; Then add the pure water of 3～8 times of Lumbricus weight, above-mentioned Lumbricus is carried out after cell breakage with pearl mill, colloidal mill or high pressure homogenizer, add the sodium citrate of above-mentioned parts by weight and Chinese medicine composite powder to carry out fully stirring and making homogenate.3) by step 2) homogenate that obtains is by filtering or the centrifugal thick slag of removing wherein of sedimentation centrifuge, homogenate after slagging-off, adopt per minute 6000 to leave centrifugal 16～24 minutes of scheming or 0.35 micron pore size membrane filtration processing, obtain clear liquor, described clear liquor is every milliliter of liquid that is greater than 180 enzyme activity units; 4) saltout: step 3) add ammonium sulfate by 35% saturated concentration in gained clear liquor, stir leave standstill 50 minutes after dissolving completely mixed liquor; 5) centrifugal ultrafiltration: with vertical high speed centrifugal machine through high speed centrifugation by step 4) in mixed liquor thin slag remove, make clear liquid, then pass through 50000dt membrane ultrafiltration, obtain ultrafiltrate.6) chromatographic column separates: adopt anion-exchange chromatography post or affinity column separating step 5) ultrafiltrate that obtains, with petroleum ether, 0.005～1.0 mole of arginine or sodium chloride solution eluting, collect the chromatography effluent of Peak Activity part; 7) sterilization: by above-mentioned steps 6) chromatography effluent put into the sealed sterilised container of carrying pipe, in container, inject high pressure N through conduit
2gas, machinery fully stirs pending clear liquor 5～8min simultaneously, through conduit output release pressure, N after pressure decreased
2dissolubility reduces rapidly, a large amount of N in container
2the rapid loss of gas; Then filter removal of contamination, precipitate 10 hours and obtain supernatant; 8) by step 7) lyophilization of gained supernatant, in subzero 28～35 degree freezer dryers freezing 5～8 hours, then, under the vacuum of 0.2mmHg, be warming up to 25～30 DEG C, dry 8 hours; Continue to be warming up to 35～39 DEG C, vacuum drying 10～12 hours, obtains powdery product, every milligram of 2000～3000 enzyme activity units.Described Abelmoschus esculentus is Abelmoschus esculentus stem and leaf, flower and the seed after cleaning up and drying.
Preferably, a kind of blood sugar lowering, blood fat reducing, blood pressure lowering, improve microcirculation, eliminate the earthworm protein of diabetic complication, it is characterized in that, described Lumbricus is selected from: day stannum Dula earthworm, Japanese Dula earthworm, Pheretima aspergillum, Pheretima pingi, Amythas dancatala, flat greatly No. two earthworms.
Preferably, a kind of blood sugar lowering, blood fat reducing, blood pressure lowering, improve microcirculation, eliminate the earthworm protein of diabetic complication, it is characterized in that, described Lumbricus is selected from: day stannum Dula earthworm, Japanese Dula earthworm, Pheretima aspergillum, Pheretima pingi, Amythas dancatala, flat greatly No. two earthworms; The described Lumbricus of selecting starts to feed in advance Cassia sophera Linn seed powder or Rhizoma Dioscoreae powder or Radix Et Rhizoma Fagopyri Tatarici powder or oatmeal or Abelmoschus esculentus powder for 30 days, continue to feed the raw material that is elected to be preparation earthworm protein after 30 days, described Cassia sophera Linn seed powder or Rhizoma Dioscoreae powder or Radix Et Rhizoma Fagopyri Tatarici powder or oatmeal or Abelmoschus esculentus powder are to be crushed to through super micron mill the powder that 350～500 orders are made after Cassia sophera Linn seed, Rhizoma Dioscoreae, Radix Et Rhizoma Fagopyri Tatarici, Herba bromi japonici, Abelmoschus esculentus drying.
Preferably, a kind of blood sugar lowering, blood fat reducing, blood pressure lowering, improve microcirculation, eliminate the capsule that the earthworm protein of diabetic complication is made, it is characterized in that, by weight, get 30～50 parts, the powder of above-mentioned earthworm protein, then adopt research of super-pine crush equipment processing to pulverize; Make pulverized powder grain diameter reach for subsequent use between 100～160 microns; Add again 5 parts of xylosic alcohol powders, after stirring, obtain mixed dust formulation; Described mixed dust formulation is added and in capsule, makes portable capsule.
Preferably, a kind of blood sugar lowering, blood fat reducing, blood pressure lowering, improve microcirculation, eliminate diabetic complication earthworm protein make capsule, it is characterized in that, described capsule is selected soft capsule or hard capsule.
Preferably, a kind of blood sugar lowering, blood fat reducing, blood pressure lowering, improve microcirculation, eliminate the earthworm protein of diabetic complication, it is characterized in that, by weight, get 30～50 parts, the powder of above-mentioned earthworm protein, then adopt research of super-pine crush equipment processing to pulverize; Make pulverized powder grain diameter reach for subsequent use between 120～200 microns; Add again 5 parts of xylosic alcohol powders, after stirring, obtain mixed dust formulation; By the agent of described mixed dust formulation granulation, pill, tea bag, tablet, effervescent tablet.
Preferably, a kind of blood sugar lowering, blood fat reducing, blood pressure lowering, improve microcirculation, eliminate the purposes of earthworm protein of diabetic complication, it is characterized in that, it is used to the medicament of preparation treatment or prevent diabetes.
Preferably, a kind of blood sugar lowering, blood fat reducing, blood pressure lowering, improve microcirculation, eliminate the purposes of earthworm protein of diabetic complication, it is characterized in that, it is used to food or the health product of preparation treatment or prevent diabetes.
Preferably, a kind of blood sugar lowering, blood fat reducing, blood pressure lowering, improve microcirculation, eliminate the purposes of earthworm protein of diabetic complication, it is characterized in that, it is used to the medicament of preparation treatment or the prevention nephrotic syndrome.
Described Chinese medicinal ingredients is through verifying for many years, record and test gained, although their effect is different, but choosing of each Chinese medicine is not to be arbitrarily, the applicant is according to the proportioning of the each composition of test gained for many years, be not by raw material effect simple superposition of each composition, because may there is the multiple interactivelies such as interpromoting relation in five elements, mutual reinforcement between, mutual restraint between two drugs or the mutual-detoxication between various raw material of Chinese medicine, and the applicant has selected above-mentioned raw materials accordingly according to this principle exactly, and determine the best proportioning between them, therefore, the application has significant meaning.
Beneficial effect:
It is raw material sources that the present invention selects natural animal and plant medical material, utilize the comprehensive function of each natural constituents, treatment diabetic complication, kidney syndrome, through acute toxicology and long-term toxicological test, prove human body nonhazardous effect, its drug effect is high, can improve high blood coagulation state, improves blood fat, reduce blood glucose, thrombus, dredging microcirculation, improves immunity of organisms simultaneously.
The capsule that the earthworm protein that the present invention extracts is made, selects gelatine capsule, and every packs 0.25g earthworm protein powder of the present invention into, and capsule properties and characteristics is yellow transparent shape, soluble in water.Show through safety toxicology test: LD50 is 20000mg/kg, Toxicological evaluation is nontoxic level, has no side effect.
Drink through human experiment, every day 2-3 grain, every 0.25g.In 150 people, have obesity (with hyperlipidemia), hypertension, hyperglycemia symptom patient, everyone drinks 30-60 days, and drug withdrawal 30-60 days.Wherein obesity patient's (with hyperlipidemia symptom) 50 people, through inspection fasting plasma lipid, T-CHOL CT, triglyceride TG and high density lipoprotein HDL, contrast test before and after drinking, carry out again curative effect comparison: test is drunk 30 days, experimenter TC, TG significantly decline, HDL obviously rises, and TC declines respectively 32.6%, 43.1%, 55.2%;
Wherein 50 hypertension symptom persons, drank after 30 days, obtained good blood pressure lowering effect simultaneously, and mean blood pressure declines 30%, blood pressure drops amplitude: systolic pressure decline 3.16kpa, and diastolic pressure decline 2.61%kpa, its hypotensive effect is remarkable;
50 hyperglycemic patients are also obtained to good blood sugar lowering, lipid-lowering effect; Experimenter drank after 30 days, the TC 1.20mmol/L that on average declines, HDL rising 0.80mmol/L, the effect of TC and rising HDL wherein obviously declines, its effect is higher than vitamin E, the average rate of decrease that plays its glucose is 12.4%, and the average rate of decrease of fructosamine is to check its blood glucose value all to reach normal level after 26.6%, 30 day.Result of the test shows: the health product effect the present invention relates to is remarkable.
Show through pharmacological testing: it is raw material sources that the present invention selects natural animal and plant medical material, utilize the comprehensive function of each Chinese medicinal components, treatment diabetic syndrome and nephrotic syndrome, through acute toxicology and long-term toxicological test, prove human body nonhazardous effect, its drug effect is high, can improve high blood coagulation state, improves blood fat, reduce blood glucose, thrombus, dredging microcirculation, improves immunity of organisms simultaneously.
Specific embodiment
Embodiment 1
Cassia sophera Linn Semen Cassiae, pulse family Cassia sophera Cassia sophera L..
Rhizoma Dioscoreae, common name Rhizoma Dioscoreae.Herbaceos perennial, stem is overgrow, and is often with purple, and tuber is cylindrical, and leaf is to life, avette or oval, flower milky, dioecism.Starch-containing and the protein of tuber.Rhizoma Dioscoreae-medicinal part, plants name: Rhizoma Dioscoreae (y ù), Latin language formal name used at school: Dioscorea opposita another name: Rhizoma Dioscoreae, triangle, RHIIZOMA DIOSCOREAE fr0m Henan of China, Rhizoma dioscoreae, medicated egg, native potato, mountain potato, jade prolong.Abnormal smells from the patient: (root) is sweet, warm, flat, nontoxic.English name: nagaimo, yamaimo, Chinese yam, Japanese mountain yam, Korean yam; Compendium of Materia Medica " to record, Rhizoma Dioscoreae is because the Tang Dynasty ancestor is Li Yu, for taboo changes Rhizoma Dioscoreae into, again because Song Yingzong is Zhao Shu, for taboo changes Rhizoma Dioscoreae into.Rhizoma Dioscoreae is one of composition medicine of LIUWEI DIHUANG WAN.[1] Rhizoma Dioscoreae " book on Chinese herbal medicine is looked for the truth ": " enter in drugs for nourishing yin and should give birth to use, entering spleen reinforcing lung medicine should use by parching to brown." " this genus food, though gas is warm and flat, is the moon of spleen reinforcing lung.Can moisten fur, longue meat, sweet in the mouth is held concurrently salty, again can kidney tonifying reinforcing YIN-essence.", in the research of C-II induced arthritis in rats, Rhizoma Dioscoreae water extraction liquid can significantly suppress the Denaturation of Cu2+ to gamma Globulin, shows to have antarthritic effect.Rhizoma Dioscoreae water extraction liquid also can be eliminated urine protein, mutant cell is had to the effect that suppresses generation.Invigorating the spleen and replenishing QI: Rhizoma Dioscoreae is planted trace element because being rich in 18 seed amino acids and more than 10, and other mineral, so have strengthening the spleen and stomach, tonifying the lung kidney, invigorating the spleen and replenishing QI, spleen invigorating tonify deficiency, the effects such as beneficial essence, beneficial heart and tranquilizing mind of reinforcing the kidney, in Li Shizhen (1518-1593 A.D.) Compendium of Material Medica, there is saying of " all kinds of diseases and ailments are reinforced the kidney, controlled to spleen invigorating tonification, taste essence, treats five kinds of strain and seven kinds of impairment ".Quench one's thirst and promote the production of body fluid: this Rhizoma Dioscoreae has effect of the life of quenching one's thirst: Chinese traditional treatment asthenia is quenched one's thirst in (diabetes) prescription often has Rhizoma Dioscoreae single to use, or share with other medicines, and effect is better.Health care: because fresh Rhizoma dioscoreae is rich in multivitamin, aminoacid and mineral, can prevent and treat Human Lipid Metabolism abnormal, and arteriosclerosis, to safeguarding that insulin normal function also has certain effect, there is enhancing body immunity, beneficial heart and tranquilizing mind, rather cough the health-care effects such as Dingchuan, slow down aging.Skin care: Yuan Dynasty taste expert Li Gao says: " control xerosis cutis and moisten it with this thing." Li Shizhen (1518-1593 A.D.) is write: " Rhizoma Dioscoreae can be moistened fur." Rhizoma Dioscoreae is to nourishes rough skin, and vigorous and graceful skin care has unique curative effect.
From effect, Cassia sophera Linn, Rhizoma Dioscoreae are all good at chronic diseases such as adjuvant therapy of diabetes complication.Earthworm protein product is more single in the market, substantially there is no the precedent of earthworm protein and Chinese medicine Application of composite, belongs to technological gap.The present invention, through long-term test and clinical verification, utilizes the compound earthworm protein of Chinese medicine to make oral liquid.Product meets the growing health care demand of people, future market promise well.In clinical practice, further verified that in prescription, various medicines share by proportioning, the single result of use of comparing obviously improves.
Other Chinese medicinal ingredients that relate to are through verifying for many years, record and test gained, although their effect is different, choosing of each Chinese medicine is not to be arbitrarily that length is limit, and does not carry out exhaustive at this;
Blood sugar lowering, blood fat reducing, blood pressure lowering, improve microcirculation, eliminate the earthworm protein of diabetic complication, it is characterized in that being prepared by following steps:
1) Lumbricus is cleaned: select Lumbricus alive, remove impurity and the rotten Lumbricus of dead, it is 4 water 10 minutes that Lumbricus is put into pH value, adds 50g Sal simultaneously, and illumination simultaneously 2 hours impels Lumbricus get rid of in body dirt and clean Lumbricus body surface; Lumbricus alive after cleaning is carried out to drip washing, use the warm water washing of 28 DEG C, wash away excessive moisture and the impurity of outer surface; 2) slurrying: by step 1) Lumbricus that obtains is positioned in container after weighing, meter is put into 30 parts by weight, pH value is 5.0 10 parts of sodium citrates, puts into 10 parts of Chinese medicine composite powders again, and described Chinese medicine composite powder is: 5 parts of 10 parts of Cassia sophera Linn seeds, 3 parts of Semen Vaccariae, 3 parts of the Radixs Astragali, 2 parts of Rhizoma Dioscoreaes, 2 parts, the Radix Rehmanniae, 2 parts of Fructus Schisandrae Chinensis, 1 part of Radix Ophiopogonis, 1 part of Fructus Mume, 1 part of Rhizoma Polygonati, 1.5 parts of Fructus Lyciis, 1 part of Radix Puerariae, 1 part, Folium Mori, Abelmoschus esculentus are crushed to 350～500 orders through super micron mill and make powder and stir and be composited; Then add the pure water of 3～8 times of Lumbricus weight, above-mentioned Lumbricus is carried out after cell breakage with pearl mill, colloidal mill or high pressure homogenizer, add the sodium citrate of above-mentioned parts by weight and Chinese medicine composite powder to carry out fully stirring and making homogenate.3) by step 2) homogenate that obtains is by filtering or the centrifugal thick slag of removing wherein of sedimentation centrifuge, homogenate after slagging-off, adopt per minute 6000 to leave centrifugal 16 minutes of scheming or 0.35 micron pore size membrane filtration processing, obtain clear liquor, described clear liquor is every milliliter of liquid that is greater than 180 enzyme activity units; 4) saltout: step 3) add ammonium sulfate by 35% saturated concentration in gained clear liquor, stir leave standstill 50 minutes after dissolving completely mixed liquor; 5) centrifugal ultrafiltration: with vertical high speed centrifugal machine through high speed centrifugation by step 4) in mixed liquor thin slag remove, make clear liquid, then pass through 50000dt membrane ultrafiltration, obtain ultrafiltrate.6) chromatographic column separates: adopt anion-exchange chromatography post or affinity column separating step 5) ultrafiltrate that obtains, with petroleum ether, 0.005 mole of arginine or sodium chloride solution eluting, collect the chromatography effluent of Peak Activity part; 7) sterilization: by above-mentioned steps 6) chromatography effluent put into the sealed sterilised container of carrying pipe, in container, inject high pressure N through conduit
2gas, machinery fully stirs pending clear liquor 5min simultaneously, through conduit output release pressure, N after pressure decreased
2dissolubility reduces rapidly, a large amount of N in container
2the rapid loss of gas; Then filter removal of contamination, precipitate 10 hours and obtain supernatant; 8) by step 7) lyophilization of gained supernatant, in subzero 28 degree freezer dryers freezing 5 hours, then, under the vacuum of 0.2mmHg, be warming up to 25 DEG C, dry 8 hours; Continue to be warming up to 35 DEG C, vacuum drying 10 hours, obtains powdery product, every milligram of 2000～3000 enzyme activity units.Described Abelmoschus esculentus is Abelmoschus esculentus stem and leaf, flower and the seed after cleaning up and drying.
Wherein chromatographic column separates and adopts Mono-Q or Q-agarose gel H.P chromatography, carries out eluting with 0.005～1.0 mole of arginine.Or adopt carboxymethyl-or DEAE-glucose gel or agarose gel column chromatography, with 1.0～1.5 mole nacls gradient elution separation go out the different earthworm protein of molecular weight.
Preferably, a kind of blood sugar lowering, improve microcirculation, eliminate the earthworm protein of complication, described Lumbricus is selected from: day stannum Dula earthworm, Japanese Dula earthworm, Pheretima aspergillum, Pheretima pingi, Amythas dancatala, flat greatly No. two earthworms.
Preferably, a kind of blood sugar lowering, blood fat reducing, blood pressure lowering, improve microcirculation, eliminate the earthworm protein of diabetic complication, it is characterized in that, described Lumbricus is selected from: day stannum Dula earthworm, Japanese Dula earthworm, Pheretima aspergillum, Pheretima pingi, Amythas dancatala, flat greatly No. two earthworms; The described Lumbricus of selecting starts to feed in advance Cassia sophera Linn seed powder or Rhizoma Dioscoreae powder or Radix Et Rhizoma Fagopyri Tatarici powder or oatmeal or Abelmoschus esculentus powder for 30 days, continue to feed the raw material that is elected to be preparation earthworm protein after 30 days, described Cassia sophera Linn seed powder or Rhizoma Dioscoreae powder or Radix Et Rhizoma Fagopyri Tatarici powder or oatmeal or Abelmoschus esculentus powder are to be crushed to through super micron mill the powder that 350～500 orders are made after Cassia sophera Linn seed, Rhizoma Dioscoreae, Radix Et Rhizoma Fagopyri Tatarici, Herba bromi japonici, Abelmoschus esculentus drying.
Preferably, a kind of blood sugar lowering, blood fat reducing, blood pressure lowering, improve microcirculation, eliminate the capsule that the earthworm protein of diabetic complication is made, it is characterized in that, by weight, get 30～50 parts, the powder of above-mentioned earthworm protein, then adopt research of super-pine crush equipment processing to pulverize; Make pulverized powder grain diameter reach for subsequent use between 100～160 microns; Add again 5 parts of xylosic alcohol powders, after stirring, obtain mixed dust formulation; Described mixed dust formulation is added and in capsule, makes portable capsule.
Preferably, a kind of blood sugar lowering, blood fat reducing, blood pressure lowering, improve microcirculation, eliminate diabetic complication earthworm protein make capsule, it is characterized in that, described capsule is selected soft capsule or hard capsule.
Preferably, a kind of blood sugar lowering, blood fat reducing, blood pressure lowering, improve microcirculation, eliminate the earthworm protein of diabetic complication, it is characterized in that, by weight, get 30～50 parts, the powder of above-mentioned earthworm protein, then adopt research of super-pine crush equipment processing to pulverize; Make pulverized powder grain diameter reach for subsequent use between 120～200 microns; Add again 5 parts of xylosic alcohol powders, after stirring, obtain mixed dust formulation; By the agent of described mixed dust formulation granulation, pill, tea bag, tablet, effervescent tablet.
Preferably, a kind of blood sugar lowering, blood fat reducing, blood pressure lowering, improve microcirculation, eliminate the purposes of earthworm protein of diabetic complication, it is characterized in that, it is used to the medicament of preparation treatment or prevent diabetes.
Preferably, a kind of blood sugar lowering, blood fat reducing, blood pressure lowering, improve microcirculation, eliminate the purposes of earthworm protein of diabetic complication, it is characterized in that, it is used to food or the health product of preparation treatment or prevent diabetes.
Preferably, a kind of blood sugar lowering, blood fat reducing, blood pressure lowering, improve microcirculation, eliminate the purposes of earthworm protein of diabetic complication, it is characterized in that, it is used to the medicament of preparation treatment or the prevention nephrotic syndrome.
Acute toxicity test:
Experimental subject: 10 of 20-23g healthy mices, male and female half and half
Experimental agents: the oral liquid that earthworm protein dry powder of the present invention is made
Test method: the dosage according to 50 times of human body administration, 100 times gavages to white mice, negative control group gives the normal saline of matched doses, animal is freely taken the photograph life and the death condition that diseases caused by retention of fluid food is observed animal, administration after 10 days is slaughtered the white mice of survival, get the variation of the indices of its its peripheral blood of whole blood assay, take to improve karber's method (Journal of the Fourth Military Medical University, the 7th phase in 2002) and ask LD
50for 20000mg/kg, Toxicological evaluation is nontoxic level, has no side effect.
The capsule that the earthworm protein that the present invention extracts is made, selects gelatine capsule, and every packs 0.25g earthworm protein powder of the present invention into, and capsule properties and characteristics is yellow transparent shape, soluble in water.Show through safety toxicology test: LD50 is 20000mg/kg, Toxicological evaluation is nontoxic level, has no side effect.
Drink through human experiment, every day 2-3 grain, every 0.25g.In 150 people, have obesity (with hyperlipidemia), hypertension, hyperglycemia symptom patient, everyone drinks 30-60 days, and drug withdrawal 30-60 days.Wherein obesity patient's (with hyperlipidemia symptom) 50 people, through inspection fasting plasma lipid, T-CHOL CT, triglyceride TG and high density lipoprotein HDL, contrast test before and after drinking, carry out again curative effect comparison: test is drunk 30 days, experimenter TC, TG significantly decline, HDL obviously rises, and TC declines respectively 32.2%, 43.3%, 55.4%;
Earthworm protein treatment diabetes: blood plasma GMP140 is one of activation releasing product of platelet or endotheliocyte, the mensuration that research shows GMP140 content in blood plasma also can antimer in hematoblastic activation degree and thrombophilia, and the metabolic end product that TXB2 is XA2, the latter has very strong short platelet aggregation and contracting blood vessel function.Many reports show that diabetics blood plasma GMP140 and TXB2 level significantly increase, and illustrate in diabetics body that platelet is in the state of activation.Platelet adhesion aggregation capability increases, and this is that diabetics arteriosclerosis occurs early and incidence rate is high and one of the reason of easy concurrent microangiopathies.Earthworm protein is to like to win from artificial breeding's compatriot the one group of acidic protein extracting Lumbricus.After the oral earthworm protein of diabetics 2 weeks, blood plasma TXB2 level obviously declines, and with matched group without significant difference, prompting earthworm protein is assembled and is had obvious inhibitory action platelet adhesion.After blood plasma GMP140 horizontal stretcher, also significantly decline, explanatorily dragon protein has the effect that suppresses Concentration In Platelets of Diabetic Patients activation to a certain extent.
The earthworm protein treatment nephrotic syndrome: earthworm protein is the histone hydrolytic enzyme extracting from extraordinary Lumbricus.The unit Preliminary Results such as Tsing-Hua University in 1984 explicitly dragon protein have thrombolytic, anticoagulant and go fine effect.Confirm at present its mechanism of action, the one, there is activator of plasminogen (similar t-PA) effect; The 2nd, be combined fibrin is degraded rapidly with fibrin.There is direct and indirect thrombolytic function.Levels in Nephrotic Syndrome Patients often has the blood coagulation activity of interior coagulation system to strengthen.Fibrinogen increases and impels blood coagulation, impels microthrombusis in glomerular capillary.NS patient's majority is in hypercoagulability, and has thrombotic tendency.The result of study proof fibrinolytic kinase activities (being mainly t-PA) such as the attached Hou Ming of institute of Shandong medical university reduce and fibrinolytic suppresses active (being mainly a2-PA) enhancing, are one of major reasons of the formation of chronic glomerulus disease high blood coagulation state and progress.Test inspection prompting fibrinogen and the blood viscosity index of this group significantly increase, and this conforms to substantially with more domestic scholars' report.Imagine the effect that this group is used earthworm protein treatment PNS, just be to supplement or increased the content of t-PA in blood, thereby in blood fibrinolytic system, activate with the imbalance state that suppresses factor and be able to rebalancing while making NS, play the effect that anticoagulant and thrombolytic improve Microcirculation of Renal Glomeruli, reach the object that reduces urine protein and improve renal function.The U.S. former permanent earth-worm extractive as raw material that uses of nineteen eighty-three Japan's Miyazaki University, the thousands of examples of clinical treatment thromboembolic disorders, effective percentage reaches more than 80%.Within 1993, the long company of Zhu waits reported 9 cases primary glomerulopathy, and after oral earthworm protein extract, twenty-four-hour urine protein quantification is treated front obvious minimizing, and 2 routine renal insufficiency persons have clear improvement after earthworm protein treatment.Therapeutic effect is challenging.The routine PNS of this group 20 total effective rate after treatment is 95%, and apparently higher than 65% of matched group, P < 0.05 has significant difference.Difference as for I type and II type curative effect may be long with its course of disease, and the degree of renal damage is heavier, and histological type difference is relevant.Because many reasons fails to do the research of pathological and Relationships Between Curative, need further to be explored.
Embodiment 2
Blood sugar lowering, blood fat reducing, blood pressure lowering, improve microcirculation, eliminate the earthworm protein of diabetic complication, it is characterized in that being prepared by following steps:
1) Lumbricus is cleaned: select Lumbricus alive, remove impurity and the rotten Lumbricus of dead, it is 4.9 water 12.5 minutes that Lumbricus is put into pH value, adds 50g Sal simultaneously, and illumination simultaneously 2.5 hours impels Lumbricus get rid of in body dirt and clean Lumbricus body surface; Lumbricus alive after cleaning is carried out to drip washing, use the warm water washing of 28 DEG C, wash away excessive moisture and the impurity of outer surface; 2) slurrying: by step 1) Lumbricus that obtains is positioned in container after weighing, meter is put into 40 parts by weight, pH value is 5.0～5.3 10～20 parts of sodium citrates, puts into 20 parts of Chinese medicine composite powders again, and described Chinese medicine composite powder is: 5 parts of 10 parts of Cassia sophera Linn seeds, 3 parts of Semen Vaccariae, 3 parts of the Radixs Astragali, 2 parts of Rhizoma Dioscoreaes, 2 parts, the Radix Rehmanniae, 2 parts of Fructus Schisandrae Chinensis, 1 part of Radix Ophiopogonis, 1 part of Fructus Mume, 1 part of Rhizoma Polygonati, 1.5 parts of Fructus Lyciis, 1 part of Radix Puerariae, 1 part, Folium Mori, Abelmoschus esculentus are crushed to 350～500 orders through super micron mill and make powder and stir and be composited; Then add the pure water of 5 times of Lumbricus weight, above-mentioned Lumbricus is carried out after cell breakage with pearl mill, colloidal mill or high pressure homogenizer, add the sodium citrate of above-mentioned parts by weight and Chinese medicine composite powder to carry out fully stirring and making homogenate.3) by step 2) homogenate that obtains is by filtering or the centrifugal thick slag of removing wherein of sedimentation centrifuge, homogenate after slagging-off, adopt per minute 6000 to leave centrifugal 20 minutes of scheming or 0.35 micron pore size membrane filtration processing, obtain clear liquor, described clear liquor is every milliliter of liquid that is greater than 180 enzyme activity units; 4) saltout: step 3) add ammonium sulfate by 35% saturated concentration in gained clear liquor, stir leave standstill 50 minutes after dissolving completely mixed liquor; 5) centrifugal ultrafiltration: with vertical high speed centrifugal machine through high speed centrifugation by step 4) in mixed liquor thin slag remove, make clear liquid, then pass through 50000dt membrane ultrafiltration, obtain ultrafiltrate.6) chromatographic column separates: adopt anion-exchange chromatography post or affinity column separating step 5) ultrafiltrate that obtains, with petroleum ether, 0.005～1.0 mole of arginine or sodium chloride solution eluting, collect the chromatography effluent of Peak Activity part; 7) sterilization: by above-mentioned steps 6) chromatography effluent put into the sealed sterilised container of carrying pipe, in container, inject high pressure N2 gas through conduit, machinery fully stirs pending clear liquor 6.5min simultaneously, through conduit output release pressure, after pressure decreased, N2 dissolubility reduces rapidly, the rapid loss of a large amount of N2 gases in container; Then filter removal of contamination, precipitate 10 hours and obtain supernatant; 8) by step 7) lyophilization of gained supernatant, in subzero 30 degree freezer dryers freezing 6.5 hours, then, under the vacuum of 0.2mmHg, be warming up to 27 DEG C, dry 8 hours; Continue to be warming up to 37 DEG C, vacuum drying 11 hours, obtains powdery product, every milligram of 2000～3000 enzyme activity units.Described Abelmoschus esculentus is Abelmoschus esculentus stem and leaf, flower and the seed after cleaning up and drying.
Wherein chromatographic column separates and adopts Mono-Q or Q-agarose gel H.P chromatography, carries out eluting with 0.008 mole of arginine.Or adopt carboxymethyl-or DEAE-glucose gel or agarose gel column chromatography, with 1.0～1.5 mole nacls gradient elution separation go out the different earthworm protein of molecular weight.
Preferably, a kind of blood sugar lowering, improve microcirculation, eliminate the earthworm protein of complication, described Lumbricus is selected from: day stannum Dula earthworm, Japanese Dula earthworm, Pheretima aspergillum, Pheretima pingi, Amythas dancatala, flat greatly No. two earthworms.
Preferably, a kind of blood sugar lowering, blood fat reducing, blood pressure lowering, improve microcirculation, eliminate the earthworm protein of diabetic complication, it is characterized in that, described Lumbricus is selected from: day stannum Dula earthworm, Japanese Dula earthworm, Pheretima aspergillum, Pheretima pingi, Amythas dancatala, flat greatly No. two earthworms; The described Lumbricus of selecting starts to feed in advance Cassia sophera Linn seed powder or Rhizoma Dioscoreae powder or Radix Et Rhizoma Fagopyri Tatarici powder or oatmeal or Abelmoschus esculentus powder for 30 days, continue to feed the raw material that is elected to be preparation earthworm protein after 30 days, described Cassia sophera Linn seed powder or Rhizoma Dioscoreae powder or Radix Et Rhizoma Fagopyri Tatarici powder or oatmeal or Abelmoschus esculentus powder are to be crushed to through super micron mill the powder that 350～500 orders are made after Cassia sophera Linn seed, Rhizoma Dioscoreae, Radix Et Rhizoma Fagopyri Tatarici, Herba bromi japonici, Abelmoschus esculentus drying.
Preferably, a kind of blood sugar lowering, blood fat reducing, blood pressure lowering, improve microcirculation, eliminate the capsule that the earthworm protein of diabetic complication is made, it is characterized in that, by weight, get 30～50 parts, the powder of above-mentioned earthworm protein, then adopt research of super-pine crush equipment processing to pulverize; Make pulverized powder grain diameter reach for subsequent use between 100～160 microns; Add again 5 parts of xylosic alcohol powders, after stirring, obtain mixed dust formulation; Described mixed dust formulation is added and in capsule, makes portable capsule.
Preferably, a kind of blood sugar lowering, blood fat reducing, blood pressure lowering, improve microcirculation, eliminate diabetic complication earthworm protein make capsule, it is characterized in that, described capsule is selected soft capsule or hard capsule.
Preferably, a kind of blood sugar lowering, blood fat reducing, blood pressure lowering, improve microcirculation, eliminate the earthworm protein of diabetic complication, it is characterized in that, by weight, get 30～50 parts, the powder of above-mentioned earthworm protein, then adopt research of super-pine crush equipment processing to pulverize; Make pulverized powder grain diameter reach for subsequent use between 120～200 microns; Add again 5 parts of xylosic alcohol powders, after stirring, obtain mixed dust formulation; By the agent of described mixed dust formulation granulation, pill, tea bag, tablet, effervescent tablet.
Preferably, a kind of blood sugar lowering, blood fat reducing, blood pressure lowering, improve microcirculation, eliminate the purposes of earthworm protein of diabetic complication, it is characterized in that, it is used to the medicament of preparation treatment or prevent diabetes.
Preferably, a kind of blood sugar lowering, blood fat reducing, blood pressure lowering, improve microcirculation, eliminate the purposes of earthworm protein of diabetic complication, it is characterized in that, it is used to food or the health product of preparation treatment or prevent diabetes.
Preferably, a kind of blood sugar lowering, blood fat reducing, blood pressure lowering, improve microcirculation, eliminate the purposes of earthworm protein of diabetic complication, it is characterized in that, it is used to the medicament of preparation treatment or the prevention nephrotic syndrome.
The capsule that the earthworm protein that the present invention extracts is made, selects gelatine capsule, and every packs 0.25g earthworm protein powder of the present invention into, and capsule properties and characteristics is yellow transparent shape, soluble in water.Show through safety toxicology test: LD50 is 20000mg/kg, Toxicological evaluation is nontoxic level, has no side effect.
Drink through human experiment, every day 2-3 grain, every 0.25g.In 150 people, have obesity (with hyperlipidemia), hypertension, hyperglycemia symptom patient, everyone drinks 30-60 days, and drug withdrawal 30-60 days.Wherein obesity patient's (with hyperlipidemia symptom) 50 people, through inspection fasting plasma lipid, T-CHOL CT, triglyceride TG and high density lipoprotein HDL, contrast test before and after drinking, carry out again curative effect comparison: test is drunk 30 days, experimenter TC, TG significantly decline, HDL obviously rises, and TC declines respectively 32.4%, 43.2%, 55.1%;
Embodiment 3
Blood sugar lowering, blood fat reducing, blood pressure lowering, improve microcirculation, eliminate the earthworm protein of diabetic complication, it is characterized in that being prepared by following steps:
1) Lumbricus is cleaned: select Lumbricus alive, remove impurity and the rotten Lumbricus of dead, it is 5.9 water 15 minutes that Lumbricus is put into pH value, adds 50g Sal simultaneously, and illumination simultaneously 3 hours impels Lumbricus get rid of in body dirt and clean Lumbricus body surface; Lumbricus alive after cleaning is carried out to drip washing, use the warm water washing of 28 DEG C, wash away excessive moisture and the impurity of outer surface; 2) slurrying: by step 1) Lumbricus that obtains is positioned in container after weighing, meter is put into 50 parts by weight, pH value is 5.3 20 parts of sodium citrates, puts into 30 parts of Chinese medicine composite powders again, and described Chinese medicine composite powder is: 5 parts of 10 parts of Cassia sophera Linn seeds, 3 parts of Semen Vaccariae, 3 parts of the Radixs Astragali, 2 parts of Rhizoma Dioscoreaes, 2 parts, the Radix Rehmanniae, 2 parts of Fructus Schisandrae Chinensis, 1 part of Radix Ophiopogonis, 1 part of Fructus Mume, 1 part of Rhizoma Polygonati, 1.5 parts of Fructus Lyciis, 1 part of Radix Puerariae, 1 part, Folium Mori, Abelmoschus esculentus are crushed to 500 orders through super micron mill and make powder and stir and be composited; Then add the pure water of 8 times of Lumbricus weight, above-mentioned Lumbricus is carried out after cell breakage with pearl mill, colloidal mill or high pressure homogenizer, add the sodium citrate of above-mentioned parts by weight and Chinese medicine composite powder to carry out fully stirring and making homogenate.3) by step 2) homogenate that obtains is by filtering or the centrifugal thick slag of removing wherein of sedimentation centrifuge, homogenate after slagging-off, adopt per minute 6000 to leave centrifugal 24 minutes of scheming or 0.35 micron pore size membrane filtration processing, obtain clear liquor, described clear liquor is every milliliter of liquid that is greater than 180 enzyme activity units; 4) saltout: step 3) add ammonium sulfate by 35% saturated concentration in gained clear liquor, stir leave standstill 50 minutes after dissolving completely mixed liquor; 5) centrifugal ultrafiltration: with vertical high speed centrifugal machine through high speed centrifugation by step 4) in mixed liquor thin slag remove, make clear liquid, then pass through 50000dt membrane ultrafiltration, obtain ultrafiltrate.6) chromatographic column separates: adopt anion-exchange chromatography post or affinity column separating step 5) ultrafiltrate that obtains, with petroleum ether, 0.005～1.0 mole of arginine or sodium chloride solution eluting, collect the chromatography effluent of Peak Activity part; 7) sterilization: by above-mentioned steps 6) chromatography effluent put into the sealed sterilised container of carrying pipe, in container, inject high pressure N through conduit
2gas, machinery fully stirs pending clear liquor 5～8min simultaneously, through conduit output release pressure, N after pressure decreased
2dissolubility reduces rapidly, a large amount of N in container
2the rapid loss of gas; Then filter removal of contamination, precipitate 10 hours and obtain supernatant; 8) by step 7) lyophilization of gained supernatant, in subzero 35 degree freezer dryers freezing 8 hours, then, under the vacuum of 0.2mmHg, be warming up to 30 DEG C, dry 8 hours; Continue to be warming up to 39 DEG C, vacuum drying 10～12 hours, obtains powdery product, every milligram of 2000～3000 enzyme activity units.Described Abelmoschus esculentus is Abelmoschus esculentus stem and leaf, flower and the seed after cleaning up and drying.
Wherein chromatographic column separates and adopts Mono-Q or Q-agarose gel H.P chromatography, carries out eluting with 0.005～1.0 mole of arginine.Or adopt carboxymethyl-or DEAE-glucose gel or agarose gel column chromatography, with 1.0～1.5 mole nacls gradient elution separation go out the different earthworm protein of molecular weight.
Preferably, a kind of blood sugar lowering, improve microcirculation, eliminate the earthworm protein of complication, described Lumbricus is selected from: day stannum Dula earthworm, Japanese Dula earthworm, Pheretima aspergillum, Pheretima pingi, Amythas dancatala, flat greatly No. two earthworms.
Preferably, a kind of blood sugar lowering, blood fat reducing, blood pressure lowering, improve microcirculation, eliminate the earthworm protein of diabetic complication, it is characterized in that, described Lumbricus is selected from: day stannum Dula earthworm, Japanese Dula earthworm, Pheretima aspergillum, Pheretima pingi, Amythas dancatala, flat greatly No. two earthworms; The described Lumbricus of selecting starts to feed in advance Cassia sophera Linn seed powder or Rhizoma Dioscoreae powder or Radix Et Rhizoma Fagopyri Tatarici powder or oatmeal or Abelmoschus esculentus powder for 30 days, continue to feed the raw material that is elected to be preparation earthworm protein after 30 days, described Cassia sophera Linn seed powder or Rhizoma Dioscoreae powder or Radix Et Rhizoma Fagopyri Tatarici powder or oatmeal or Abelmoschus esculentus powder are to be crushed to through super micron mill the powder that 350～500 orders are made after Cassia sophera Linn seed, Rhizoma Dioscoreae, Radix Et Rhizoma Fagopyri Tatarici, Herba bromi japonici, Abelmoschus esculentus drying.
Preferably, a kind of blood sugar lowering, blood fat reducing, blood pressure lowering, improve microcirculation, eliminate the capsule that the earthworm protein of diabetic complication is made, it is characterized in that, by weight, get 30～50 parts, the powder of above-mentioned earthworm protein, then adopt research of super-pine crush equipment processing to pulverize; Make pulverized powder grain diameter reach for subsequent use between 100～160 microns; Add again 5 parts of xylosic alcohol powders, after stirring, obtain mixed dust formulation; Described mixed dust formulation is added and in capsule, makes portable capsule.
Preferably, a kind of blood sugar lowering, blood fat reducing, blood pressure lowering, improve microcirculation, eliminate diabetic complication earthworm protein make capsule, it is characterized in that, described capsule is selected soft capsule or hard capsule.
Preferably, a kind of blood sugar lowering, blood fat reducing, blood pressure lowering, improve microcirculation, eliminate the earthworm protein of diabetic complication, it is characterized in that, by weight, get 30～50 parts, the powder of above-mentioned earthworm protein, then adopt research of super-pine crush equipment processing to pulverize; Make pulverized powder grain diameter reach for subsequent use between 120～200 microns; Add again 5 parts of xylosic alcohol powders, after stirring, obtain mixed dust formulation; By the agent of described mixed dust formulation granulation, pill, tea bag, tablet, effervescent tablet.
Preferably, a kind of blood sugar lowering, blood fat reducing, blood pressure lowering, improve microcirculation, eliminate the purposes of earthworm protein of diabetic complication, it is characterized in that, it is used to the medicament of preparation treatment or prevent diabetes.
Preferably, a kind of blood sugar lowering, blood fat reducing, blood pressure lowering, improve microcirculation, eliminate the purposes of earthworm protein of diabetic complication, it is characterized in that, it is used to food or the health product of preparation treatment or prevent diabetes.
Preferably, a kind of blood sugar lowering, blood fat reducing, blood pressure lowering, improve microcirculation, eliminate the purposes of earthworm protein of diabetic complication, it is characterized in that, it is used to the medicament of preparation treatment or the prevention nephrotic syndrome.
The capsule that the earthworm protein that the present invention extracts is made, selects gelatine capsule, and every packs 0.25g earthworm protein powder of the present invention into, and capsule properties and characteristics is yellow transparent shape, soluble in water.Show through safety toxicology test: LD50 is 20000mg/kg, Toxicological evaluation is nontoxic level, has no side effect.
Drink through human experiment, every day 2-3 grain, every 0.25g.In 150 people, have obesity (with hyperlipidemia), hypertension, hyperglycemia symptom patient, everyone drinks 30-60 days, and drug withdrawal 30-60 days.Wherein obesity patient's (with hyperlipidemia symptom) 50 people, through inspection fasting plasma lipid, T-CHOL CT, triglyceride TG and high density lipoprotein HDL, contrast test before and after drinking, carry out again curative effect comparison: test is drunk 30 days, experimenter TC, TG significantly decline, HDL obviously rises, and TC declines respectively 32.5%, 43.2%, 55.1%;
The numerical range relating in the embodiment of being described herein is all through many experiments gained, and each point can be realized, and is not confined to end points or terminal, and length is limit, and does not enumerate at this.
Finally it should be noted that: above embodiment only, in order to technical scheme of the present invention to be described, is not intended to limit; Although the present invention is had been described in detail with reference to previous embodiment, those of ordinary skill in the art is to be understood that: its technical scheme that still can record aforementioned each embodiment is modified, or part technical characterictic is wherein equal to replacement; And these amendments or replacement do not make the essence of appropriate technical solution depart from the spirit and scope of various embodiments of the present invention technical scheme.
Claims (1)
1. blood sugar lowering, blood fat reducing, blood pressure lowering, improve microcirculation, eliminate diabetic complication earthworm protein make a capsule, it is characterized in that being prepared by following steps:
1) Lumbricus is cleaned: select Lumbricus alive, remove impurity and the rotten Lumbricus of dead, it is 4～5.9 water 10～15 minutes that Lumbricus is put into pH value, adds 50g Sal simultaneously, the 2-3 hour of illumination simultaneously, impels Lumbricus get rid of the interior dirt of body and clean Lumbricus body surface; Lumbricus alive after cleaning is carried out to drip washing, use the warm water washing of 28 DEG C, wash away excessive moisture and the impurity of outer surface; 2) slurrying: by step 1) Lumbricus that obtains is positioned in container after weighing, meter is put into 30～50 parts by weight, pH value is 5.0～5.3 10～20 parts of sodium citrates, puts into 10～30 parts of Chinese medicine composite powders again, and described Chinese medicine composite powder is: 5 parts of 10 parts of Cassia sophera Linn seeds, 3 parts of Semen Vaccariae, 3 parts of the Radixs Astragali, 2 parts of Rhizoma Dioscoreaes, 2 parts, the Radix Rehmanniae, 2 parts of Fructus Schisandrae Chinensis, 1 part of Radix Ophiopogonis, 1 part of Fructus Mume, 1 part of Rhizoma Polygonati, 1.5 parts of Fructus Lyciis, 1 part of Radix Puerariae, 1 part, Folium Mori, Abelmoschus esculentus are crushed to 350～500 orders through super micron mill and make powder and stir and be composited; Then add the pure water of 3～8 times of Lumbricus weight, above-mentioned Lumbricus is carried out after cell breakage with pearl mill, colloidal mill or high pressure homogenizer, add the sodium citrate of above-mentioned parts by weight and Chinese medicine composite powder to carry out fully stirring and making homogenate; 3) by step 2) homogenate that obtains is by filtering or the centrifugal thick slag of removing wherein of sedimentation centrifuge, homogenate after slagging-off, adopt per minute 6000 to leave centrifugal 16～24 minutes of scheming or 0.35 micron pore size membrane filtration processing, obtain clear liquor, described clear liquor is every milliliter of liquid that is greater than 180 enzyme activity units; 4) saltout: step 3) add ammonium sulfate by 35% saturated concentration in gained clear liquor, stir leave standstill 50 minutes after dissolving completely mixed liquor; 5) centrifugal ultrafiltration: with vertical high speed centrifugal machine through high speed centrifugation by step 4) in mixed liquor thin slag remove, make clear liquid, then pass through 50000dt membrane ultrafiltration, obtain ultrafiltrate; 6) chromatographic column separates: adopt anion-exchange chromatography post or affinity column separating step 5) ultrafiltrate that obtains, with petroleum ether, 0.005～1.0 mole of arginine or sodium chloride solution eluting, collect the chromatography effluent of Peak Activity part; 7) sterilization: by above-mentioned steps 6) chromatography effluent put into the sealed sterilised container of carrying pipe, in container, inject high pressure N through conduit
2gas, machinery fully stirs pending clear liquor 5～8min simultaneously, through conduit output release pressure, N after pressure decreased
2dissolubility reduces rapidly, a large amount of N in container
2the rapid loss of gas; Then filter removal of contamination, precipitate 10 hours and obtain supernatant; 8) by step 7) lyophilization of gained supernatant, in subzero 28～35 degree freezer dryers freezing 5～8 hours, then, under the vacuum of 0.2mmHg, be warming up to 25～30 DEG C, dry 8 hours; Continue to be warming up to 35～39 DEG C, vacuum drying 10～12 hours, obtains powdery product, every milligram of 2000～3000 enzyme activity units; Described Abelmoschus esculentus is Abelmoschus esculentus stem and leaf, flower and the seed after cleaning up and drying;
Wherein chromatographic column separates and adopts Mono-Q or Q-agarose gel H.P chromatography, carries out eluting with 0.005～1.0 mole of arginine; Or adopt carboxymethyl-or DEAE-glucose gel or agarose gel column chromatography, with 1.0～1.5 mole nacls gradient elution separation go out the different earthworm protein of molecular weight;
Described Lumbricus is selected from: day stannum Dula earthworm, Japanese Dula earthworm, Pheretima aspergillum, Pheretima pingi, Amythas dancatala, flat greatly No. two earthworms; The described Lumbricus of selecting starts to feed in advance Cassia sophera Linn seed powder or Rhizoma Dioscoreae powder or Radix Et Rhizoma Fagopyri Tatarici powder or oatmeal or Abelmoschus esculentus powder for 30 days, continue to feed the raw material that is elected to be preparation earthworm protein after 30 days, described Cassia sophera Linn seed powder or Rhizoma Dioscoreae powder or Radix Et Rhizoma Fagopyri Tatarici powder or oatmeal or Abelmoschus esculentus powder are to be crushed to through super micron mill the powder that 350～500 orders are made after Cassia sophera Linn seed, Rhizoma Dioscoreae, Radix Et Rhizoma Fagopyri Tatarici, Herba bromi japonici, Abelmoschus esculentus drying;
Described capsule by weight, is got 30～50 parts, the powder of above-mentioned earthworm protein, then adopts research of super-pine crush equipment processing to pulverize; Make pulverized powder grain diameter reach for subsequent use between 100～160 microns; Add again 5 parts of xylosic alcohol powders, after stirring, obtain mixed dust formulation; Described mixed dust formulation is added and in capsule, makes portable capsule.
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| CN201310173902.0A CN103239706B (en) | 2013-05-13 | 2013-05-13 | Earthworm protein for reducing blood glucose, improving microcirculation and eliminating diabetic complication and application thereof |
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| CN109420155A (en) * | 2017-08-24 | 2019-03-05 | 熊小利 | Hypoglycemic improves ground dragon protein and its application that diabetic complication is eliminated in microcirculation |
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| CN107736607A (en) * | 2017-10-30 | 2018-02-27 | 中盐工程技术研究院有限公司 | A kind of earthworm subtracts sodium seasoning salt and preparation method thereof |
| CN110870520A (en) * | 2018-09-04 | 2020-03-10 | 天津星宇航天生物科技有限公司 | A nutraceutical product suitable for improving microcirculation of astronauts |
| CN116035218A (en) * | 2023-02-03 | 2023-05-02 | 海南华研胶原科技股份有限公司 | Pea oligopeptide health care product and preparation method thereof |
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