Studies On Compatibility Principal Of HED-effects And Mechanism Of The Compatibility Of HED Anti-inflammation And Anti-allergic Asthma

Herba Ephedrae Decoction (HED) is composed of monarch drug Herba Ephedrae, ministerial drug Romulus Cinnamomi, adjuvant drug Semen Armeniacae Amarum and cionductant drug Radix Glycyrrhizae. The compatibility of HED is very right-sizingm, elaborative. The Chinese herbalist doctors have had the identical understanding about the compatibility and pharmacology of HED for the past Dynasties. Therefore, in this paper, HED was selected to as the best representation for the traditional Chinese compositions to be studied for the compatibility principles of prescriptions.Objectives: To study the effects and mechanisms of HED on anti-inflammatory and antianaphylaxis asthma on the level of organ, cell and molecule so as to analyze the effects of ministerial drug Ramulus Cinnamomi, adjuvant drug Semen Armeniacae Amarum and cionductant drug Radix Glycyrrhizae on the pharmacology of monarch drug Herba Ephedrae in HED, and further to explore the compatibility rule of HED, which could provide the experiences for references of methodology and enrich the theories of compatibility rules of Traditional Chinese Medicines.Methods: Having employing the orthogonal design L₈(2⁷) table to device the experimental groups, we considered the fixed Herba Ephedrae as the fixed factor, and Ramulus Cinnamomi, Semen Armeniacae Amarum and Radix Glycyrrhizae as the other three factors, "yes" and "no" as two levels for each factor, eight groupscontaining Herba Ephedrae as fllow: (D Herba Ephedrae (9g),Ramulus Cinnamomi(6g), Amreniacae Amarum(6g), Glycyrrhizae(3g);(2) Herba Ephedrae (9g),Ramulus Cinnamomi(6g), Amreniacae Amarum(6g); (3) Herba Ephedrae (9g),Ramulus Cinnamomi(6g), Glycyrrhizae(3g); ? erba Ephedrae (9g),Ramulus Cinnamomi(6g); (5) Herba Ephedrae (9g), Amreniacae Amarum(6g), Glycyrrhizae(3g);(g) Herba Ephedrae (9g), Amreniacae Amarum(6g); (7) Herba Ephedrae (9g) Glycyrrhizae(3g);(8)Herba Ephedrae (9g).Every group was added 240 ml water and marinated 30 min , Herba Ephedrae (9g)was dropped at first and decocted 20 min, then other drugs were dropped and decocted 30min, filtrated by gauze and concentrated to 18ml(0.5g/ml calculated by containing Herba Ephedrae). and All the experiments were carried out as follows: (1) investigating the acute toxicity of HED in mice by using Bliss method, antianaphylaxis action of HED via the experiment of mast cell degranule induced by OA in rat, anti-inflammatory action of HED through the experiment of enhancement of capillary vessel permeability induced by acetic acid in mice and ear swelling induced by dimethyl benzene in mice, which could draw a conclusion of toxicity, antianaphylaxis and anti-inflammatory of HED, (2) constructing the anaphylactic asthma model induced by OA : Mice were immunised on days 0 and 14 by intraperitoneal (i.p.) injection of 10 (xg of ovalbumin, in 0.2 ml of saline with 20 mg of aluminium hydroxide. From day 21 the animals were challenged with aerosolised ovalbumin (5% in saline) or vehicle (saline) for 30 minutes per day on six consecutive days. Aerosol generation was achieved by use of an air-driven nebuliser The eosinophils aggregation in bronchoalveolar lavage fluid (BALF) and peripheral blood in the ovalbum in-sensitized mice after challenged with antigen in vivo after orally administered HED and the other separated compositions in mice and the granulation of the peritoneal mast cells in the ovalbum in-sensitized rats after challenge in vitro were observed. (3) Neutrophil aggregation was induced by in-traperitoneal injection ofglycogen in rats.,intra peritoneal lavage fluid(PLF)was collectedandneu-trophils were removed.Neutrophils were stimulated by calciumionophore A23187in vitro to produce leukotriene B4.The concentrationsof leukotriene B4 was measured by reversed-phase high-performance li-quid chromatography(HPLC) ,Chemotatic chamber assay was used to investigate the regulative role of MHD on chenmotaxis of the neutrophils in response to LPS stimulation., (4) establishing the asthma model induced by OA in mice and then administered orally the HED and separated compositions to these mice, employing the real-tme fluorescence quantitative PCR method to measure the contents of FLAP mRNA and IL-4mRNA expressions in lung and using the ELISA method to determine the content of LTC4 in the washing solution of pulmonary alveolus and bronchi.Results: (1) The HED could obviously inhibit the mast cell degranule in rat. Meanwhile, it could significantly oppress the enhancement of capillary vessel permeability and the ear swelling in mice, which had no visible dose-effect relationship. In addition, the LD50 of HED was 19.2 g/kg.(2) In the inducing asthma mice, the invasion of acidophilia granulocyte in the washing solution of lung and in the peripheral blood were markedly increased. The HED could reduce the invasion of eosinophils in bronchoalveolar lavage fluid and in the peripheral blood, which was different from the separated compositions of HED. Compared the model group, the composition of Herba Ephedrae, Amreniacae Amarum, Glycyrrhizae and the composition of Herba Ephedrae Glycyrrhizae; the composition of Herba Ephedrae had no significant difference;Compared with the HD, the composition of Herba Ephedrae,Ramulus Cinnamomi, Amreniacae Amarum and the composition of Herba Ephedrae,Ramulus Cinnamomi, Glycyrrhizae and the composition of Herba Ephedrae,Ramulus Cinnamomi had no no significant difference. By the way, the HED and the separated compositions also drestrained the mastocyte degranule and ear swelling in rat to some various degrees.(3) The HED and the separated compositions could control the chemotaxis of neutrophils induced by LPS to different extent. Furthermore, they could repress the LTB release in the different degrees, which the HED, the composition of Herba Ephedrae, Ramulus Cinnamomi, Semen Armeniacae Amarum and the composition of Herba Ephedrae, Ramulus Cinnamomi, Radix Glycyrrhizae was evidently superior to the other separated compositions and indicated that Herba Ephedrae and Ramulus Cinnamomi had synergetic action.(4) In the lung of asthma mice, the expressions of FLAP and IL-4 and the content of LTC4 was significantly augmented compared with the control group. The HED and the separated compositions could suppress the expressions of FLAP and IL-4 and LTC4 release to a great extent in mice. The gene expression of FLAP increased significantly in model group, compared in groups, the HED, the composition of Herba Ephedrae, Ramulus Cinnamomi, Semen Armeniacae Amarum and the composition of Herba Ephedrae, Ramulus Cinnamomi, Radix Glycyrrhizae had no significant difference; the composition of Herba Ephedrae, Ramulus Cinnamomi, Semen Armeniacae Amarum and the composition of Herba Ephedrae, Ramulus Cinnamomi, Radix Glycyrrhizae had no significant difference with the composition of Herba Ephedrae, Ramulus Cinnamom. The gene expression of IL-4 increased significantly in model group, the HED, the composition of Herba Ephedrae, Ramulus Cinnamomi, Semen Armeniacae Amarum and the composition of Herba Ephedrae, Ramulus Cinnamomi, Radix Glycyrrhizae had no significant difference; the composition of Herba Ephedrae, Ramulus Cinnamomi and the composition of Herba Ephedrae, Semen Armeniacae Amarum, Radix Glycyrrhizae , the composition of Herba Ephedrae, Semen Armeniacae Amarum, the composition of Herba Ephedrae, Radix Glycyrrhizae ,and the composition of Herba Ephedrae had no significant difference. The production of LTC4 increased significantly in model group, The HED and the separated compositions could suppress theproduction of LTC4, However, Herba Ephedrae, the composition of Ramulus Cinnamomi and Radix Glycyrrhizae, the composition of Herba Ephedrae, Semen Armeniacae Amarum had no significant difference with the model group.Conclusions: The HED had the remarkable effects of anti-inflammatory and antianaphylaxis asthma. The mechanism of HED antianaphylaxis asthma may be correlated with suppressing inflammation and stabilizing mast cell membrane.The results through separating the compositions of HED in this study suggested that the original HED was the best composition for anti-inflammatory and antianaphylaxis. At the moment, the effects of original HED had no significant differences compared with the composition of Herba Ephedrae, Ramulus Cinnamomi, Semen Armeniacae Amarum and the composition of Herba Ephedrae, Ramulus Cinnamom, Radix Glycyrrhizae. On the other hand, the pharmacological effects of composition of Herba Ephedrae and Ramulus Cinnamomi was better than those of the composition of Herba Ephedrae, Semen Armeniacae Amarum and the composition of Herba Ephedrae, Radix Glycyrrhizae, which showed that Ramulus Cinnamomi and Herba Ephedrae had notable synergetic effects on anti-inflammatory and antianaphylaxis and that composing Herba Ephedrae as monarch drug, Ramulus Cinnamomi as ministerial drug in the HED was reasonable. In brief, the present paper demonstrated the scientificaalness and reasonableness of the composition of HED in vivo, in vitro including the experiments on organ, cell and molecular level.