CN100349589C - Medical application of rehmannia root extractive in resisting asthma and allergy - Google Patents
Medical application of rehmannia root extractive in resisting asthma and allergy Download PDFInfo
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Abstract
The present invention relates to medicine for treating and preventing asthma diseases, chronic obstructive pulmonary diseases and allergy disease symptoms, which is prepared by extracting traditional Chinese medicine (rehmannia root). The extract of the rehmannia root and the total glycoside of the rehmannia root have obvious anti-inflammatory activity and obvious anti-anaphylaxis activity, and can be used as natural medicine for treating and preventing asthma diseases chronic obstructive pulmonary diseases and allergy disease symptoms. The present invention has good therapeutic effect, and has no toxic or side effect of the existing anti-asthma medicine.
Description
Technical field
The present invention relates to field of medicaments, a kind of medical usage that effectively prevents and treat the natural plant extracts of asthma, chronic obstructive pulmonary disease (COPD), irritated disease.The present invention is specially adapted to treatment and prevention of asthma.Also can be used for treatment and Polyglucan rhinitis and anaphylaxis dermatosis.
Background technology
Asthma is the respiratory tract anaphylaxis reactive inflammation disease that allergen causes.Allergen is the key factor that causes pathogenesis of asthma and development, and allergen is deposited on the air flue mucosa after sucking air flue, causes airway allergy inflammation by local and systemic immune response.Sucking allergen causes airway allergy inflammation branch sensitization and causes scorching two stages.Atopy patient produces sensitization after beginning to suck certain allergen, and body can be sensitiveness to this allergen.In the sensitization stage, it is that air flue produces and the corresponding specificity antibody IgE of allergen that body mainly changes, mastocyte in this antibody IgE and the air flue, acidophil and hugely have a liking for cell affinity is all arranged, these interior cells of these specificity antibody IgEs and air flue combine the sensitization state that has formed air flue and body.Can cause activation and release inflammatory mediators such as mastocyte when identical allergen sucks air flue once more, cause air flue allergenicity inflammation and cause asthma attack, Here it is causes the scorching stage.It also is the pathogenic process of asthma.
Bronchial asthma is chronic respiratory tract disease the most common in the world wide, and the conservative estimation whole world has at least the people more than 100,000,000 to suffer from asthma, in recent years, and because of its prevalence and mortality rate all are taking advantage of a favourable situation of rising.Asthma has become serious public health problem and has received the very big concern of countries in the world.Therefore need provide the medicine and the method for the treatment of asthma effectively.
In the last thirty years, a large amount of development and researches has been carried out to the medicine of treatment asthma in the whole world.The medicine of the treatment asthma that has developed has β
2-3 adrenergic receptor agonists class, alpha-adrenergic aceptor antagonist class, aminophylline class, cholinoceptor antagonist class, calcium ion antagonist class, potassium-channel agonist, glucocorticoid, cell membrane stability agent (sodium cromoglicate, Buddhist nun examine a meter sodium more), antihistamine drug, LTRA, platelet activating factor (PAF) antagonist, specific active immunotherapy agent, nonspecific immunity therapeutic agent etc.Asthma is the very complicated multi-factor disease of a kind of mechanism, is the diseases associated with inflammation that the many cells multimedium participates in.Above-mentioned these medicines (perhaps Therapeutic Method) mechanism of action single (to a kind of or two, three kind of medium, the factor, receptor or passage work), these medicines or therapeutic effect not obvious or oral invalid must the special pathway administration or side effect greatly can't life-time service.What generally adopt clinically at present also is that the most effective Therapeutic Method is to suck glucocorticoid (light moderate sucks sodium cromoglicate, the Buddhist nun examines a meter sodium more) anti-inflammatory treatment for a long time, sucks β during asthma attack
2Bronchodilators such as-3 adrenergic receptor agonists, cholinoceptor antagonist.These Therapeutic Method are to relieving asthma symptoms, and there is certain effect the quality of life of improving asthma patient, but the M ﹠ M of asthma is decreased.Reduce the M ﹠ M of asthma, the prognosis that improves asthma still lack effective means (Li Minghua etc., asthma, the People's Health Publisher, 1998, p2).The treatment asthma needs more effective and safer medicine.
The cause of disease of chronic obstructive pulmonary disease is at present also not clear, but relevant with airway inflammation be sure, use long-term glucocorticoid anti-inflammatory treatment and the suction β of sucking at present
2The treatment of bronchodilators such as-3 adrenergic receptor agonists, cholinoceptor antagonist is that certain effect is arranged.But the therapeutic effect of these medicines is not very good, and the medicine of seeking more effective treatment chronic obstructive pulmonary disease is very important.
Summary of the invention
The present invention relates to treat the Chinese medicine pharmaceutical composition of asthma and other anaphylactic disease, is the natural medicinal formulations of its effective ingredient with the iridoid glycoside that is contained in the Chinese medicine Radix Rehmanniae particularly.
Radix Rehmanniae is fresh tuber or the dry tuber of scrophulariaceae rehmannia glutinosa plant Rehmannia glutinosa Libsch., and perennial herb is mainly the cultivation product, and is also wild in the hillside and the wasteland, roadside of height above sea level 50-1100 rice.Be distributed in ground such as Liaoning, the Inner Mongol, Hebei, Henan, Shanxi, Shaanxi, Shandong, Anhui, Zhejiang, Hubei, Hunan, Sichuan.The document record contains number of chemical compositions such as iridoid glycosides, monoterpene and glycosides thereof, phenolic glycoside, organic acid and organic acid esters glycosides, polysaccharide, aminoacid, sterol in the Radix Rehmanniae, reach 20 kinds of metallic elements such as calcium, magnesium, potassium, ferrum, zinc, manganese.
Existing pharmacopeia record, Radix Rehmanniae has the branch of Radix Rehmanniae, Radix Rehmanniae, Radix Rehmanniae Preparata, and Radix Rehmanniae has clearing away heat and promoting production of body fluid, removing heat from blood, hemostatic effect.Be used for the hot blast impairment of YIN, the crimson tongue excessive thirst is sent out the speckle dermexanthesis, spits blood epistaxis, laryngopharynx swelling and pain.Radix Rehmanniae has clearing away heat and cooling blood, yin nourishing, the effect of promoting the production of body fluid.Be used for calentura crimson tongue excessive thirst, interior-heat caused by deficiency of YIN, the hectic fever due to YIN-deficiency consumptive fever, interior-heat is quenched one's thirst, and spits blood, and epistaxis is sent out the speckle dermexanthesis.Radix Rehmanniae Preparata has nourishing YIN and supplementing blood, and beneficial essence is filled out the effect of marrow.Be used for the hepatic and renal YIN deficiency, soreness of the waist and knees, osteopyrexia and fever, the night sweat seminal emission, interior-heat is quenched one's thirst, blood deficiency and yellow complexion, severe palpitation, menoxenia, metrostaxis, dizzy, tinnitus, early whitening of beard and hair.All there is not the record of asthma, antiallergic effect.Be used to prepare the treatment asthma, treat the Radix Rehmanniae of the Radix Rehmanniae extract of chronic obstructive pulmonary disease, anaphylactic disease, be after Radix Rehmanniae, Radix Rehmanniae and Radix Rehmanniae natural drying product (dry or air-dry) or Radix Rehmanniae boil 5-10 minute (killing enzyme) with boiling water, to dry or air-dry dry product.Be used to prepare the treatment asthma, treat the Radix Rehmanniae of the Radix Rehmanniae total glucosides extract use of chronic obstructive pulmonary disease, anaphylactic disease, be after Radix Rehmanniae, Radix Rehmanniae and Radix Rehmanniae natural drying product (dry or air-dry) or Radix Rehmanniae boil 5-10 minute (killing enzyme) with boiling water, to dry or air-dry dry product.
The purpose of this invention is to provide the application of Radix Rehmanniae extract in the pharmaceutical preparation of preparation treatment asthma, treatment chronic obstructive pulmonary disease, anaphylactic disease and diseases associated with inflammation.Radix Rehmanniae extract extracts from the Chinese medicine Radix Rehmanniae, comprises water extract, alcohol extract, the water-alcohol extraction of Radix Rehmanniae, and the material that extracts from Radix Rehmanniae with other solvent.Especially, be meant the Radix Rehmanniae total glucosides extract that Radix Rehmanniae iridoids chemical constituent is an effective ingredient that contains that from Radix Rehmanniae, extracts, the Radix Rehmanniae total glucosides extract mainly is made up of following chemical constituent: catalpol (catalpol), Martynoside. A, B, C, D (rehmannioside A, B, C, D), acetylcatalpol (acetylcatalpol), dihydro catalpol (dihydrocatalpol etc., and other iridoid constituents, phenolic glycoside and other micro constitutent, and be main effective ingredient with catalpol.Described Radix Rehmanniae total glucosides extract total glycosides content can be not less than 50%, and catalpol content can be not less than 20%.
Characteristics such as Radix Rehmanniae extract is a kind of novel have treatment asthma, treatment chronic obstructive pulmonary disease, active plant extract of antiallergic pharmacology, and it is little to have toxicity, and the life-time service curative effect is preferable.
The Radix Rehmanniae total glucosides extract is a kind of novel have asthma, active plant extract of antiallergic pharmacology, and it is little to have a dosage, characteristics such as toxicity is little, and the life-time service curative effect is preferable.
The Wistar healthy mice is used in the Radix Rehmanniae extract acute toxicity test, and oral maximum dosage-feeding is equivalent to not see animal dead more than the Radix Rehmanniae crude drug 150g/kg; Intraperitoneal injection LD
50Be equivalent to crude drug 50/kg, intravenous administration LD
50Be 20～30g/kg.
Iridoid glycoside content is greater than 50%, and catalpol content is done acute toxicity test greater than 20% Radix Rehmanniae total glucosides extract with the Wistar healthy mice, and oral maximum dosage-feeding is 56.5g/kg, does not see animal dead; Intraperitoneal injection LD
50Be 18.5～25.5g/kg, intravenous administration LD
50Be 2.47～3.15g/kg.
Radix Rehmanniae total glucosides provided by the invention can mix with one or more pharmaceutically acceptable stone compositions, and contains in the compositions of Radix Rehmanniae total glucosides and further contain β
2-3 adrenergic receptor agonists.
The invention provides the method that the treatment mammal breathes the immune-mediated inflammatory diseases in road, described method comprises the Radix Rehmanniae total glucosides extract to described administration treatment effective dose, or use the pharmaceutical composition that contains the Radix Rehmanniae total glucosides extract, or further contain β
2The described compositions of-3 adrenergic receptor agonists.
The pharmaceutical composition of Radix Rehmanniae total glucosides extract can be a various forms, comprises peroral dosage form, the solution of suction form and injectable and infusion.When using with inhalant or aerosol form, the Radix Rehmanniae total glucosides extract is used in combination with the dry powder that pharmaceutically acceptable carrier solution maybe can change into the form of aerosol.Similarly, when using with oral administration, the Radix Rehmanniae total glucosides extract is used in combination with the pharmaceutically acceptable carrier that is used in oral administration.When being used for the treatment of immune-mediated inflammatory dermatosis, the Radix Rehmanniae total glucosides extract can be used in combination with nontoxic, pharmaceutically acceptable topical vehicle.The Radix Rehmanniae total glucosides extract can with the treating asthma agent, as β
2-3 adrenergic receptor agonists, cholinoceptor antagonist, aminophylline class, glucocorticoid, antihistamine drug etc. are used in combination.
The Radix Rehmanniae total glucosides extract is used to prevent and treat immune-mediated inflammatory diseases, particularly comprises the disease relevant with respiratory tract of asthma, and particularly relevant with chronic asthma replying by force and allergic rhinitis.Therefore the present invention also provides the method for the treatment of immune-mediated inflammatory diseases, and this method comprises to the Radix Rehmanniae total glucosides extract that has immune-mediated inflammatory diseases patient with treatment effective dosage forms and dosage.
The dosage of Radix Rehmanniae extract of the present invention, because of patient's state, body weight, administering mode etc. different different.Such as under non-oral situation, intramuscular, vein, enteral administration, calculate 0.20mg-2000mg/kg. day with the Radix Rehmanniae crude drug, preferred 2.0mg-200mg/kg. day, calculate 2.0-20g/kg. day with the Radix Rehmanniae crude drug under the oral situation, preferred 20-2000mg/kg. day.
Iridoid glycoside content of the present invention is greater than 50%, and catalpol content is greater than the dosage of 20% Radix Rehmanniae total glucosides extract, because of patient's state, body weight, administering mode etc. different different.Such as under non-oral situation, intramuscular, vein, enteral administration 0.01mg-100mg/kg. day, preferred 0.1mg-10mg/kg. day, 0.1-1000mg/kg. day under the oral situation, preferred 1-100mg/kg. day.
The specific embodiment
With following embodiment the present invention is specified, but the present invention is not limited to the content that the following example comprises.
[embodiment one] gets Radix Rehmanniae 20kg, water boils for 160 liters and extracted 1 hour, leaching extracting solution, medicinal residues add 120 liters in water again and boil extraction 2 hours, leaching extracting solution, medicinal residues add 120 liters in water again and boil extraction 2 hours, the leaching extracting solution merges aqueous extract, vacuum concentration to 20 liter, get Radix Rehmanniae extracting solution, place refrigerator to preserve.
The clinical research of Radix Rehmanniae extracting solution treatment asthma
1, case is selected 60 examples, male 30 examples, and women 30 examples, age 16-60 year, (People's Health Publisher, 1998, standard diagnostics p165) was the bronchial asthma patient according to chief editor'ss " asthma " such as Li Minghua.Be divided into 2 groups at random, every group 30 example, male 15 examples, women 15 examples; The A group is the treatment group, and B is a matched group.2, Therapeutic Method, the treatment group is oral with Radix Rehmanniae extracting solution, and every day 3 times, each 8ml took 30 days continuously; The matched group propionic acid beclometasone aerosol, every day 2 times, each 200 μ g took 30 days continuously.3, therapeutic outcome, when treatment finished, the treatment group was cured 22 examples, effective 8 examples; Matched group is cured 10 examples, effective 20 examples.Treatment finishes back 1 month, and the treatment group is cured 14 examples, effective 13 examples; Matched group is cured 5 examples, effective 18 examples.
[embodiment two] get Radix Rehmanniae 1kg, water 8000ml boils and extracted 1 hour, the leaching extracting solution, medicinal residues add water 6000ml again and boil extraction 1 hour, and leaching extracting solution, medicinal residues add water 6000ml again and boil extraction 1 hour, the leaching extracting solution, merge aqueous extract, vacuum concentration gets Radix Rehmanniae crude extract (1) 430g to doing.The ovalbumin aerosol sucks to draw breathes heavily test
Get 60 of 200-250g Cavia porcelluss, male and female half and half, prior to right rear leg outside intramuscular injection 4% ovalbumin normal saline solution 0.2ml/ only, while lumbar injection 4% gel aluminum hydroxide 0.2ml sensitization.From sensitization second day, animal was divided into 2 groups at random: matched group, treatment group (4.0g/kg).Treatment group Radix Rehmanniae crude extract (1) 4.0g/kg gastric infusion, totally 14 days, behind the last medicine, animal is put in the airtight glass bell jar, after treating peace and quiet, start air compressor, spray into 30 seconds of 3.5% ovalbumin normal saline, observe incubation period and the number of animals that tic takes place that the tic of panting property appears in Cavia porcellus in 6 minutes with the constant voltage of 53kPa (400mmHg).(Xu Shuyun, Bian Rulian, Chen Xiu chief editor, pharmacological experimental methodology, the People's Health Publisher, in November, 1991 second edition, 1199 pages).
Table 1, Radix Rehmanniae crude extract (1) are drawn the influence that Cavia porcellus breathes heavily to medicine
| Group | Number of animals (n) | Dosage (g/kg) | Draw and breathe heavily incubation period (second) | Tic number of animals (only) |
| Contrast Radix Rehmanniae crude extract (1) | 10 10 | - 4.0 | 62.8±31.7 149.5±97.8* | 10/10 9/10 |
Annotate: relatively think significant difference in P＜0.05, (representing) with * with the blank group.
Result of the test shows, Radix Rehmanniae crude extract (1) 4.0g/kg dosage can draw the incubation period of breathing heavily Cavia porcellus by significant prolongation ovalbumin normal saline solution, compare with matched group that there were significant differences (P＜0.05), show that Radix Rehmanniae crude extract (1) has certain antiasthmatic effect.
Passive cutaneous anaphylaxis test (PCA)
The rat body is divided into 2 groups at random by heavy, 10/group, ♀ ♂ half and half, sub-cage rearing, 5/cage.Be respectively blank group, reagent group 4.0g/kg.d.Therapeutic administration Radix Rehmanniae crude extract (1) 4.0g/kg.d is adopted in test, successive administration Radix Rehmanniae crude extract (1) 14d before antigen is attacked.
Antiserum Preparation: with four sole injections of above-mentioned Radix Trichosanthis aluminium hydroxide suspension, every sole injection 0.1ml, 0.4ml altogether.The 10-15d maturation, the about 10ml of eyeball blood-letting with the centrifugal 15min of 3000 commentaries on classics/min, gets upper serum and promptly gets antiserum.
Passive sensitization of skin: will test and respectively organize the rat back depilation, 2 points are respectively got apart from center line 1.5cm in the spinal column both sides, every some interval 2cm, the about 1 * 1cm2 of area, totally 4 points.In skin of back, inject sero-fast different dilution factors (1: 30 1: 40) respectively.Carrying out quantitative corresponding antigens behind the 48h attacks.The tail vein injection Radix Trichosanthis adds AZO-blue solution 1mg/kg, sacrificed by decapitation behind the 30min.
Observation index: the sero-fast local skin reaction of intradermal injection, and measure the locus coeruleus diameter that AZO-blue is oozed out.Calculate PCA and suppress percentage rate (%).PCA suppress percentage rate=(blank group locus coeruleus diameter-medication group locus coeruleus diameter)/blank group locus coeruleus diameter * 100% (Xu Shuyun, Bian Rulian, the Chen Xiu chief editor, pharmacological experimental methodology, the People's Health Publisher, in November, 1991 second edition, 1197 pages)
Table 2, Radix Rehmanniae crude extract (1) are to models of passive skin irritability of rats reaction influence
| Group | Number of animals (only) | Dosage mg/kg | Different serum-concentration locus coeruleus diameters (cm) | |
| 1∶30 | 1∶40 | |||
| Blank Radix Rehmanniae crude extract (1) | 10 10 | Water 150.0 | 1.45±0.22 0.95±0.21* | 0.90±0.25 0.57±0.22* |
Annotate: relatively think significant difference in P＜0.05, (representing) with * with the blank group.
Result of the test sees Table 1.From table the result as can be seen, reagent Radix Rehmanniae crude extract (1) is antiserum dilution factor 1: 30,1: 40 o'clock, show significance and suppress locus coeruleus diameter (P＜0.05), suppression ratio is 33.1% and 30.%.Show that Radix Rehmanniae crude extract (1) has certain inhibitory action to passive cutaneous anaphylaxis, PCA.
[embodiment three] get Radix Rehmanniae 1kg, ethanol water 8000ml reflux, extract, with 70% 1 hour, the leaching extracting solution, medicinal residues added 70% ethanol water 6000ml reflux, extract, 1 hour again, and leaching extracting solution, medicinal residues added 70% ethanol water 6000ml reflux, extract, 1 hour again, the leaching extracting solution, merge extractive liquid,, vacuum concentration gets Radix Rehmanniae crude extract (2) 230g to doing.
The ovalbumin aerosol sucks to draw breathes heavily test
Get 60 of 200-250g Cavia porcelluss, male and female half and half, prior to right rear leg outside intramuscular injection 4% ovalbumin normal saline solution 0.2ml/ only, while lumbar injection 4% gel aluminum hydroxide 0.2ml sensitization.From sensitization second day, animal is divided into 2 groups at random: matched group, treatment group, treatment group Radix Rehmanniae crude extract (2) 2.5g/kg gastric infusion, totally 14 days, behind the last medicine, animal is put in the airtight glass bell jar, after treating peace and quiet, start air compressor, spray into 30 seconds of 3.5% ovalbumin normal saline, observe incubation period and the number of animals that tic takes place that the tic of panting property appears in Cavia porcellus in 6 minutes with the constant voltage of 53kPa (400mmHg).(chief editor such as Xu Shuyun, pharmacological experimental methodology, People's Health Publisher, second edition in 1991,1199 pages)
Table 3, Radix Rehmanniae crude extract (2) are drawn the influence that Cavia porcellus breathes heavily to medicine
| Group | Number of animals (n) | Dosage (g/kg) | Draw and breathe heavily incubation period (second) | Tic number of animals (only) |
| Contrast Radix Rehmanniae crude extract (2) | 10 10 | - 2.5 | 59.8±30.6 152.5±101.7* | 10/10 9/10 |
Annotate: relatively think significant difference in P＜0.05, (representing) with * with the blank group.
Result of the test shows, Radix Rehmanniae crude extract (2) 2.5g/kg dosage can draw the incubation period of breathing heavily Cavia porcellus by significant prolongation ovalbumin normal saline solution, compare with matched group that there were significant differences (P＜0.05), show that Radix Rehmanniae crude extract (2) has certain antiasthmatic effect.
[embodiment four] get Radix Rehmanniae 1kg, with methanol 8000ml reflux, extract, 1 hour, the leaching extracting solution, medicinal residues add methanol 6000ml again and boil extraction 1 hour, and leaching extracting solution, medicinal residues add methanol 6000ml reflux, extract, 1 hour again, the leaching extracting solution, merge extractive liquid,, vacuum concentration gets Radix Rehmanniae crude extract (3) 216g to doing.
The ovalbumin aerosol sucks to draw breathes heavily test
Get 60 of 200-250g Cavia porcelluss, male and female half and half, prior to right rear leg outside intramuscular injection 4% ovalbumin normal saline solution 0.2ml/ only, while lumbar injection 4% gel aluminum hydroxide 0.2ml sensitization.From sensitization second day, animal is divided into 2 groups at random: matched group, Radix Rehmanniae crude extract (3) group, Radix Rehmanniae crude extract (3) group Radix Rehmanniae crude extract (3) 2.5g/kg gastric infusion, totally 14 days, behind the last medicine, animal is put in the airtight glass bell jar, after treating peace and quiet, start air compressor, spray into 30 seconds of 3.5% ovalbumin normal saline, observe incubation period and the number of animals that tic takes place that the tic of panting property appears in Cavia porcellus in 6 minutes with the constant voltage of 53kPa (400mmHg).(Xu Shuyun, Bian Rulian, Chen Xiu chief editor, pharmacological experimental methodology, the People's Health Publisher, in November, 1991 second edition, 1199 pages).
Table 4, Radix Rehmanniae crude extract (3) are drawn the influence that Cavia porcellus breathes heavily to medicine
| Group | Number of animals (n) | Dosage (g/kg) | Draw and breathe heavily incubation period (second) | Tic number of animals (only) |
| Contrast Radix Rehmanniae crude extract (3) | 10 10 | - 2.5 | 59.8±30.6 152.5±101.7* | 10/10 9/10 |
Annotate: relatively think significant difference in P＜0.05, (representing) with * with the blank group.
Result of the test shows, Radix Rehmanniae crude extract (3) 2.5g/kg dosage can draw the incubation period of breathing heavily Cavia porcellus by significant prolongation ovalbumin normal saline solution, compare with matched group that there were significant differences (P＜0.05), show that Radix Rehmanniae crude extract (3) has certain antiasthmatic effect.
[embodiment five] get Radix Rehmanniae 2kg, water 16000ml boils and extracted 1 hour, the leaching extracting solution, medicinal residues add water 12000ml again and boil extraction 1 hour, and leaching extracting solution, medicinal residues add water 12000ml again and boil extraction 1 hour, the leaching extracting solution, merge aqueous extract,, get Radix Rehmanniae total glucosides 135g through further purification, HPLC, TLC detect and to show and contain catalpol (catalpol), Martynoside. A, B, C, D (rehmanniosideA, B, C, D), acetylcatalpol (acetylcatalpol), (dihydrocatalpol etc. also contain a spot of phenolic glycoside to the dihydro catalpol.Assay shows that catalpol content is 21.3%, and iridoid glycoside content is 51.4%.
The ovalbumin aerosol sucks to draw breathes heavily test
Get 60 of 200-250g Cavia porcelluss, male and female half and half, prior to right rear leg outside intramuscular injection 4% ovalbumin normal saline solution 0.2ml/ only, while lumbar injection 4% gel aluminum hydroxide 0.2ml sensitization.From sensitization second day, animal is divided into 2 groups at random: matched group, Radix Rehmanniae total glucosides group, Radix Rehmanniae total glucosides group Radix Rehmanniae total glucosides 150mg/kg gastric infusion, totally 14 days, behind the last medicine, animal is put in the airtight glass bell jar, after treating peace and quiet, start air compressor, spray into 30 seconds of 3.5% ovalbumin normal saline, observe incubation period and the number of animals that tic takes place that the tic of panting property appears in Cavia porcellus in 6 minutes with the constant voltage of 53kPa (400mmHg).(chief editor such as Xu Shuyun, pharmacological experimental methodology, People's Health Publisher, second edition in 1991,1199 pages).
Table 5, Radix Rehmanniae total glucosides draw the influence that Cavia porcellus breathes heavily to medicine
| Group | Number of animals (n) | Dosage (mg/kg) | Draw and breathe heavily incubation period (second) | Tic number of animals (only) |
| Contrast RGTG50 | 10 10 | - 150.0 | 58.7±32.3 145.5±95.3* | 10/10 9/10 |
Annotate: relatively think significant difference in P＜0.05, (representing) with * with the blank group.
Result of the test shows that Radix Rehmanniae Radix Rehmanniae total glucosides 150mg/kg dosage can draw the incubation period of breathing heavily Cavia porcellus by significant prolongation ovalbumin normal saline solution, compares with matched group that there were significant differences (P＜0.05), shows that the Radix Rehmanniae Radix Rehmanniae total glucosides has certain antiasthmatic effect.
Passive cutaneous anaphylaxis test (PCA)
The rat body is divided into 2 groups at random by heavy, 10/group, ♀ ♂ half and half, sub-cage rearing, 5/cage.Be respectively blank group, reagent group 150mg/kg.d.The Radix Rehmanniae total glucosides of therapeutic administration is adopted in test, successive administration Radix Rehmanniae total glucosides 14d before antigen is attacked.
Antiserum Preparation: with four sole injections of above-mentioned Radix Trichosanthis aluminium hydroxide suspension, every sole injection 0.1ml is total to the 0.4ml.10-15d maturation, and the about 10ml of eyeball blood-letting with the centrifugal 15min of 3000 commentaries on classics/min, gets upper serum and promptly gets antiserum.
Passive sensitization of skin: will test and respectively organize the rat back depilation, 2 points are respectively got apart from center line 1.5cm in the spinal column both sides, every some interval 2cm, the about 1 * 1cm2 of area, totally 4 points.In skin of back, inject sero-fast different dilution factors (1: 30 1: 40) respectively.Carrying out quantitative corresponding antigens behind the 48h attacks.The tail vein injection Radix Trichosanthis adds AZO-blue solution 1mg/kg, sacrificed by decapitation behind the 30min.
Observation index: the sero-fast local skin reaction of intradermal injection, and measure the locus coeruleus diameter that AZO-blue is oozed out.
Calculate PCA and suppress percentage rate (%).PCA suppresses percentage rate=(blank group locus coeruleus diameter-medication group locus coeruleus diameter)/blank group locus coeruleus diameter * 100% chief editors such as (, pharmacological experimental methodology, People's Health Publisher, second edition in 1991,1197 pages) Xu Shuyun.
Table 6, Radix Rehmanniae total glucosides are to models of passive skin irritability of rats reaction influence
| Group | Number of animals (only) | Dosage mg/kg | Different serum-concentration locus coeruleus diameters (cm) | |
| 1∶30 | 1∶40 | |||
| The blank Radix Rehmanniae total glucosides | 10 10 | Water 150.0 | 1.42±0.28 0.95±0.21* | 0.85±0.28 0.59±0.22* |
Annotate: relatively think significant difference in P＜0.05, (representing) with * with the blank group.
Result of the test sees Table 1.From table the result as can be seen, the reagent Radix Rehmanniae total glucosides is antiserum dilution factor 1: 30,1: 40 o'clock, show significance and suppress locus coeruleus diameter (P＜0.05), suppression ratio is 33.1% and 30.%.Show that reagent has certain inhibitory action to passive cutaneous anaphylaxis, PCA.
Tardy property footpad reaction test
Mice is pressed the body weight random packet, 10/group, ♀ ♂ half and half, sub-cage rearing, totally 2 groups, is respectively blank group, Radix Rehmanniae total glucosides group by 5/cage.Adopt the therapeutic administration., administration 5d continues administration 5d before the sensitization to sensitization, altogether 10d.
0.05ml/ sensitization of NIH mice cervical region s.c5% sheep red blood cell (SRBC) (SRBC) behind the sensitization 5d.Right foot pad s.c5% sheep red blood cell (SRBC) (SRBC) 0.02ml attacks behind the 6d.Left side foot injection equivalent normal saline compares.Behind the 24h, put to death animal behind the last administration 50min.Cutting two foots from the ankle joint position weighs.Obtain the weight difference of two foots, calculate the foot swelling suppression ratio.Foot swelling suppression ratio=blank group foot swelling degree-medication group foot swelling degree)/and blank group foot swelling degree * 100%, result of the test sees Table 4.(chief editor such as Xu Shuyun, pharmacological experimental methodology, People's Health Publisher, second edition in 1991,1226 pages, 1228 pages)
The influence of the tardy property of the mice footpad reaction that table 7, Radix Rehmanniae total glucosides cause sheep red blood cell (SRBC)
| Group | Number of animals (only) | Dosage (mg/kg) | Foot swelling degree (g) | Suppression ratio (%) |
| Blank group Radix Rehmanniae total glucosides | 9 10 | Water 400.0 | 2.51±0.73 1.72±0.76* | --- 31.47 |
Annotate: relatively think significant difference in P＜0.05, (representing) with * with the blank group.
From table the result as can be seen, Radix Rehmanniae total glucosides and blank group comparison sheet reveal significance inhibition foot swelling degree (P＜0.05), suppression ratio is 31.47%.Show that Radix Rehmanniae total glucosides is more obvious to the mice tardy property immunoreation effect that the inhibition sheep red blood cell (SRBC) causes.
[embodiment six] get Radix Rehmanniae 30kg, water 240000ml boils and extracted 1 hour, the leaching extracting solution, medicinal residues add water 180000ml again and boil extraction 1 hour, and leaching extracting solution, medicinal residues add water 180000ml again and boil extraction 1 hour, the leaching extracting solution, merge aqueous extract,, get Radix Rehmanniae total glucosides 2.12kg behind the vacuum drying through further purification, HPLC, TLC detects and to show and contain catalpol (catalpol), Martynoside. A, B, C, D (rehmannioside A, B, C, D), acetylcatalpol (acetylcatalpol), (dihydrocatalpol etc. also contain a spot of phenolic glycoside to the dihydro catalpol.Assay shows that catalpol content is 21.4%, and iridoid glycoside content is 51.5%.
The capsular preparation of Radix Rehmanniae total glucosides, Radix Rehmanniae total glucosides 1200 grams are sub-packed in 4000 capsules.Be Martynoside. capsule preparations of the present invention.Specification is every and contains Radix Rehmanniae total glucosides 300mg.
The clinical research of Radix Rehmanniae total glucosides treatment asthma
1, case is selected 60 examples, male 30 examples, and women 30 examples, age 16-60 year, (People's Health Publisher, 1998, standard diagnostics p165) was the bronchial asthma patient according to chief editor'ss " asthma " such as Li Minghua.Be divided into 2 groups at random, every group 30 example, male 15 examples, women 15 examples; The A group is the treatment group, and B is a matched group.2, Therapeutic Method, treatment group Radix Rehmanniae total glucosides capsule oral, each 1, was taken 30 days continuously at every day 3 times; The matched group propionic acid beclometasone aerosol, every day 2 times, each 200 μ g took 30 days continuously.3, therapeutic outcome, when treatment finished, the treatment group was cured 21 examples, effective 9 examples; Matched group is cured 12 examples, effective 18 examples.When treatment finished back 1 month, the treatment group was cured 15 examples, effective 13 examples; Matched group is cured 5 examples, effective 18 examples.4, efficacy analysis detects the pulmonary function after treatment group and the treatment of control group, the results are shown in Table 8.IgE concentration before and after the medication, the eosinophil count, numeration of leukocyte the results are shown in Table 9.
The variation of the lung function index before and after table 8, treatment group and the treatment of control group
| Group | The treatment segmentation | FEV 1(ml) | FEV 1％ | MVV(L) | PEFR(L/S) |
| The treatment group | Before taking medicine | 1877±489 | 51.67±10.12 | 63.66±16.65 | 3.80±0.67 |
| After taking medicine | 2563±583*** | 68.60±15.67*** | 83.65±18.65*** | 4.79±1.05*** | |
| Change before and after taking medicine | 686±593 | 16.93±16.45 | 19.99±18.77 | 0.99±1.11 |
| Matched group | Before taking medicine | 1879±509 | 51.21±10.19 | 62.73±16.70 | 3.80±0.75 |
| After taking medicine | 2231±589* | 63.60±15.88** | 77.65±19.75** | 4.37±1.08* | |
| Change before and after taking medicine | 352±578 | 12.39±15.96 | 14.92±20.06 | 0.57±0.97 | |
| Treatment group and matched group are relatively | * | - | - | - | |
Annotate * and represent p＜0.05; Annotate * * and represent p＜0.01; Annotate * * * and represent p＜0.001
The variation of the physiochemical indice before and after table 9, treatment group and the treatment of control group
| Group | The treatment segmentation | IgE(Iu/ml) | Acidophil | Numeration of leukocyte |
| The treatment group | Before taking medicine | 173.6±14.3 | 114±12 | 0.765±0.045 |
| After taking medicine | 78.9±9.8*** | 55±6*** | 0.735± 0.025*** | |
| Change before and after taking medicine | 94.7±15.6 | 59±13 | 0.030±0.046 | |
| Matched group | Before taking medicine | 178.6±14.8 | 117±13 | 0.758±0.035 |
| After taking medicine | 103.5± 12.3*** | 55±7*** | 0.726± 0.018*** | |
| Change before and after taking medicine | 75.1±14.7 | 62±15 | 0.032±0.042 | |
| Treatment group and matched group are relatively | *** | - | - | |
Annotate * and represent p＜0.05; Annotate * * and represent p＜0.01; Annotate * * * and represent p＜0.001
From The above results as can be seen, treatment group Radix Rehmanniae total glucosides capsule oral, every day 3 times, each 1, take 30 days therapeutic effect continuously and use propionic acid beclometasone aerosol, every day to inhale the people 2 times, each 200 μ g, the therapeutic effect of taking 30 days continuously is suitable.Some treatment index also is better than inhaling the therapeutic effect of people's propionic acid beclometasone aerosol.
Claims (1)
1, Radix Rehmanniae water extract, 70% ethanol extraction or methanolic extract are in the application that is used to prepare the Chinese medicine for the treatment of asthma, chronic obstructive pulmonary disease or allergic rhinitis.
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