The Immunopharmacological Effect And Mechanism Study Of Traditional Chinese Medicine Biminkang Granules On Allergic Rhinitis Animal Model

Purpose:The average incidence of allergic rhinitis (AR) is 10%～20%, and there are no effectivemethods and drugs for the disease. The purpose of this study is to probe into rationality offormula and dose of Biminkang granules, evaluate immunopharmacological effect, clarifymechanism, explore effective traditional Chinese medicine compound prescriptions for AR.Methods:1. Using ethology of AR mice C₅₇BL6 as index, uniform design is adopted to optimizeformula and dose of Biminkang granules.2. Using the inhibition ratio of isolated intestinal smooth muscle's allergic constriction ofguinea pigs and diameter of blue stain and optical density of OD₆₁₀ of allergic guinea pigsinduced by histamine phosphate as indexes, probe into the effect of anti histamine andimmunopharmacological effect on allergy typeâ… .3. Using the ethology, eosinophils in nasal mucosa secretion and nasal mucosa,histomorphology of mucous membrane of nose, number of mast cells and active mast cells,substance P and IgE in serum and nasal mucosa of allergic rhinitis guinea pigs as indexes,probe into the immunopharmacological effect of Biminkang granules.4. Using the level of gene expression of IL-4mRNA and NGFmRNA of allergic rhinitisguinea pigs as index, probe into the mechanism of Biminkang granules.Conclusions:1. The study on proportioning of fomula and dose.1.1 The position and interaction effects of every drug: F value indicate: the sequence ofimportance is: aconite＞astragalus＞Saposhnikovia divaricata (Turcz.) Schischk＞ephedra＞asarum herb＞atractylodes macrocephala; ephedra and asarum herb interact each other, thesame as ephedra and astragalus, astragalus and aconite, asarum herb and aconite, astragalusand atractylodes macrocephala1.2 The influence of every drug's dose on ethology of AR mice C₅₇BL6:1.2.1 The relationship of ephedra's dose and number of grabbing nose of mice: When thedose of other drugs is under designed maximum magnitude, ephedra has positive relationshipwith the number of grabbing nose,the larger of dose,the more of the number of grabbingnose.1.2.2 The relationship of aconite's dose and number of grabbing nose of mice: When thedose of other drugs is under designed maximum magnitude, aconite has negative relationshipwith the number of grabbing nose, the larger of dose, the fewer of the number of grabbingnose.1.2.3 The relationship of asarum herb's dose and number of grabbing nose of mice: Whenthe dose of other drugs is under designed maximum magnitude, asarum herb has negative relationship with the number of grabbing nose, the larger of dose, the fewer of the number ofgrabbing nose.1.2.4 The relationship of astragalus' dose and number of grabbing nose of mice: When thedose of other drugs is under designed maximum magnitude, astragalus has negativerelationship with the number of grabbing nose, the larger of dose, the fewer of the number ofgrabbing nose.1.2.5 The relationship of atractylodes macrocephala' s dose and number of grabbing noseof mice: When the dose of other drugs is under designed maximum magnitude, atractylodesmacrocephala has negative relationship with the number of grabbing nose, the larger of dose,the fewer of the number of grabbing nose.1.2.6 The relationship of Saposhnikovia divaricata (Turcz.) Schischk' s dose and number ofgrabbing nose of mice: When the dose of other drugs is under designed maximum magnitude,Saposhnikovia divaricata (Turcz.) Schischk has negative relationship with the number ofgrabbing nose, the larger of dose, the fewer of the number of grabbing nose.1.3 The influence of drug's interaction on ethology of AR mice C₅₇BL6:1.3.1 The relationship of ephedra and asarum herb: When the dose of other drugs is underdesigned maximum magnitude, ephedra has positive relationship with asarum herb. That is tosay, when ephedra and asarum herb are used together, the number of grabbing nose isdecreased distinctly even on low dose.1.3.2 The relationship of ephedra and astragalus: When the dose of other drugs is underdesigned maximum magnitude, ephedra has negative relationship with astragalus. That is tosay, when the dose of ephedra is smaller and thestragalus is larger, the number of grabbingnose is decreased.1.3.3 The relationship of astragalus and aconite: When the dose of other drugs is underdesigned maximum magnitude, ephedra has positive relationship with asarum herb. That is tosay, when astragalus and aconite are used together, the larger of dose, the fewer of number ofgrabbing nose.1.3.4 The relationship of asarum herb and aconite: When the dose of other drugs is underdesigned maximum magnitude, asarum herb has no distinctive relationship with aconite.When asarum herb and aconite are used together,the dose of asarum herb has no distinctiveeffect on the number of grabbing nose.1.3.5 The relationship of astragalus and Saposhnikovia divaricata (Turcz.) Schischk: Whenthe dose of other drugs is under designed maximum magnitude, ephedra has negativerelationship with astragalus. That is to say, when the dose of atractylodes macrocephala issmaller and the stragalus is larger, the number of grabbing nose is decreased.1.3.6 The relationship of Saposhnikovia divaricata (Turcz.) Schischk and other drugs:Saposhnikovia divaricata (Turcz.) Schischk has no interaction with other drugs.2. The effect of Biminkang granules on anti- allergy typeâ… and allergy mediators2.1 The effect of Biminkang granules on allergic constriction of isolated intestinal smoothmuscle of guinea pigs induced by ovaibumin. Constriction of isolated intestinal smooth muscleis inhibited by Biminkang granules, and display evident dose effect relationship (P＜0.01).High-dose group is similar with loratadine.2.2 The effect of Biminkang granules on permeability of skin vessels of allergic guinea piginduced by histamine phosphate: the average diameter of blue stain of high-dose, medium-dose and lower-dose experimental groups is smaller than model group (P＜0.01) and display evident dose effect relationship. High-dose groups is similar with loratadine.3. Immunopharmacological effect of Biminkang granule on anti-AR:3.1 The effect of Biminkang granules on ethology of AR guinea pigs: Symptom of AR, suchas sneezing,rhinorrhea and grabbing nose frequently were observed in model group andalmost no symptom in blank control group. Symptom of AR guinea pigs in experimentalgroup is less than model group, The difference of two groups has significant meaning (P＜0.01). The effect of Biminkang granules is better than traditional chinese medicine Xinqingranules (P＜0.05). This results indicate: Typical symptoms of AR of guinea pigs areimproved by Biminkang granules, the effect is better than that of Xinqin granules, but not asgood as loratadine.3.2 The effect of Biminkang granules on histomorphology of mucous membrane of nose ofAR guinea pigs: the epithelia of mucous membrane of nose of guinea pig in blank controlgroup are arranged in columns, eosinophil and infiltration of mast cell aren't observed underoptical microscope.The structure of epithelia in model group is abnormal, basementmembrane is destroyed and dropping, blood vessels of lamina propria are dilation andhyperemia, with much eosinophii, infiltration of mast cell and degranulation of mast cells.Eosinophil, infiltration of mast cell and degranulation of mast cells are decrease evidently inexperimental group.3.3 The effect of Biminkang granules on eosinophil in nasal secretion and mucousmembrane of nose of AR guinea pigs: the number of eosinophiles of model group andexperimental group is higher than blank control group (P＜0.01), experimental group islower than model group (P＜0.01). Biminkang granules experimental group is lower thanother traditional chinese medicine experimental group (P＜0.05) The difference betweenBiminkang granules experimental group and western medicine experimental group has nosignificant meaning (P＞0.05). This results indicate: the number of eosinophiles of ARguinea pigs is reduced by Biminkang granules, this effects is better than Xinqin granules andas good as loratadine.3.4 The effect of Biminkang granules on the number of mast cell and active mast cell ofmucous membrane of nose of AR guinea pigs: the number of mast cell and active mast cell ofmucous membrane of nose of guinea pigs of model group and experimental group is higherthan blank control group (P＜0.01), the number of mast cell and active mast cell of everyexperimental group is lower than model group (P＜0.01),the difference of mast cell betweenthe Biminkang granules experimental groups and other experimental group has no significantmeaning (P＞0.05).The number of active mast cell of Biminkang granules experimentalgroup is lower than other traditional Chinese medicine experimental group (P＜0.01) andhas no significant meaning with loratadine. The results indicate: the number of mast cell ofAR guinea pigs is decreased by Biminkang granules, the effect is similar with Xinqin granulesand ioratadine. The effect of reducing active mast cell is better than Xinqin granules, similarwith loratadine.3.5 The effect of Biminkang granules on substance P of serum of AR guinea pigs: thecontent of substance P of model group and experimental group is higher than blank controlgroup (P＜0.01), the content of every experimental group is lower than model group (P＜0.01), the content of the Biminkang granules experimental group has no significantdifference with other experimental groups (P＞0.05). The results indicate: Biminkanggranules can reduce the content of P substance, the effect is similar with Xinqin granules and loratadine.3.6 The effect of Biminkang granules on serum IgE level of allergic rhinitis mice C57BL6:the level of model group and's serum IgE is higher than blank control group's (P＜0.01),experimental group's serum IgE is lower than model group's, the level of serum IgE ofBiminkang granule's experimental group is lower than traditional chinese medicine controlgroup and western medicine control group (P＜0.01). The results indicate: Biminkanggranules can reduce the content of IgE level, The effect is better than Xinqin granules, similarwith loratadine.4. Influence of Biminkang granules on the level of gene expression of IL-4mRNA andNGFmRNA of AR guinea pigs:4.1 The effect of Biminkang granules on the level of gene expression of IL-4mRNA ofallergic rhinitis guinea pigs: the level of gene expression of IL-4mRNA of model group andevery experimental group is higher than blank control group (P＜0.01), and the level ofexperimental groups is lower than model group's (P＜0.01), the level of Biminkanggranules experimental group has significant difference with other experimental groups (P＜0.01). The results indicate: Biminkang granules can reduce the level of gene expression, Theeffect is better than Xinqin granules and loratadine.4.2 The effect of Biminkang granules on the level of gene expression of NGFmRNA ofallergic rhinitis guinea pig: the level of gene expression of NGFmRNA of model group andevery experimental group is higher than blank control group (P＜0.01), and the level ofexperimental groups is lower than model group's (P＜0.01), the level of Biminkanggranules experimental group has significant difference with other traditional Chinesemedicine experimental group (P＞0.05), but has significant difference with Ioratadine (P＜0.01). The results indicate: Biminkang granules can reduce the level of gene expression,The effect is better than Ioratadine, and similar with Xinqin granules.Conclusions:1. This article adopts uniform design method, AR mice model ,research the dose andformula of traditional Chinese medicine Biminkang granules. The results indicate: Thetherapeutic results will get the best when aconite and astragalus are used as monarch drug,Saposhnikovia divaricata (Turcz.) Schischk and ephedra as minister drug, asarum herb andatractylodes macrocephala as adjacent, and guiding drugs, the dose of aconite, astragalus,Saposhnikovia divaricata (Turcz.) Schischk, asarum herb is on designed upper-limit; ephedra,atractylodes macrocephala on designed lower-limit. The interaction of every drug is:therelationship of ephedra and asarum herb, astragalus and aconite are positive; ephedra andastragalus, astragalus and atractylodes macrocephala are negative.2. Biminkang granules can inhibit allergic constriction of isolated intestinal smooth muscleof guinea pigs, reduce permeability of skin vessels effectively.Bi min kang granules canimprove the symptoms of AR guinea pigs, decrease the number of eosinophil in nasal secretionand mucous membrane of nose, mast cell and active mast cell, the content of P substance andthe level of IgE. The mechanism is probably related with reducing level of IgE, infiltration ofeosinophil and mast cell.3. Biminkang granules can reduce level ofgene expression of IL-4mRNA and NGFmRNA ofAR guinea pigs. This is probably the main mechanism of anti-AR.4. Under clinic equivalent dosage, the effects of Biminkang granules on improving symptoms of AR, reducing the number of eosinophil and mast cell and level of gene expressionof IL-4mRNA are better than Xinqin granules. On reducing the number of eosinophil andmast cell, content of P substance, the effects are similar with loratadine. The effects ofreducing the level of IgE and gene expression of IL-4mRNA and NGFmRNA are better thanloratadine.