Pharmacological Properties of Traditional Medicine (XXX): Effects of Gyokuheifusan (玉屏風散) on Murine Antigen-Specific Antibody Production
Biological and Pharmaceutical Bulletin
In the theory of traditional Chinese medicine (TCM) theory, eqi ( ) circulates outside the vessels at the superficial portion of the body and controls the opening and closing of the pores to guard the surface of the body against exopathogen ( ). When eqi is insufficient and fails to consolidate the superficial resistance and to protect the body and skin, the exopathogens are apt to invade causing a disease such as simple flu, rhinitis and dermatitis. In TCM, this disease condition is named
... ition is named ehyo-fuko ( ), which is characterized by the symptoms of qi-deficiency ( ), aversion to wind-cold ( ), spontaneous perspiration and slow floating pulse, etc. Gyokuheifusan (GHS; , Jade Windscreen Powder in English) is a first-choice TCM formula to treat this disease condition. GHS contains three herbal medicines: root of Astragalus membranaceus BUNGE ( ), rhizome of Atractylodes ovata DECANDILLE ( ) and root of Saposhnikovia divaricata SCHISCHKIN ( ). In TCM theory, GHS invigorates qi and consolidates the superficial resistance, which increase patient's ability to protect from invasion by external pathogenic influence. 3) GHS can be used to treat biomedically defined disorders such as upper respiratory tract infection, exacerbation of chronic bronchitis, mild bronchitis, hyperthyroid condition, autonomic dystonia and allergic rhinitis. 3) In our previous study, we reported the preventive and curative effects of GHS on allergic rhinitis induced with Japanese cedar pollens in guinea pig. 2) In the pathology of TCM for pollinosis, pollens are regarded as one of exopathogens, and GHS is expected to consolidates nasal superficial resistance to escape the invasion of pollens. In our previous study, drugs had been administered to guinea pigs in the period of pollen-sensitization, and were not used just before the challenge of pollinosis by pollen inhalation. In this experimental schedule, usual anti-allergic drugs or anti-histamines could not reduce the symptoms of pollinosis, while GHS could suppress the symptoms. 2) These results have suggested that GHS could suppress the sensitization of Japanese cedar pollens to decrease IgE antibody production. Therefore, in this study, we evaluated the effects of GHS on murine antibody production using ovalbumin (OVA) as an external antigen. GHS showed different patterns for the antibody production when external antigen was injected into the inside of the body or treated on the surface of the nasal mucosa. MATERIALS AND METHODS Materials Gyokuheifusan (GHS, Lot # ACG) in the form of a dried powder extract was supplied by Iskra Industry Co., Ltd. (Tokyo). The daily dose of GHS (1.5 g) contained the extract of the mixture of following herbs: astragalus root (6 g), atractylodes rhizome (2 g), and saposhnikovia root (2 g), all of which are registered in Japanese Pharmacopoeia XIV. 4) The extract and the voucher specimens of the herbs were kept at Hunan Jiuzhitang Co, Ltd. (Changsha, Hunan, China). These medicinal herbs were boiled in their 6-times weight of H 2 O for 60 min, filtered, and then the decoction was lyophilized. Three-dimensional HPLC chromatogram of GHS and the contents of marked compounds are shown in our previous report. 2) Ovalbumin (OVA) and bovine serum albumin (BSA) were purchased from Seikagaku Corporation (Tokyo) and Sigma Aldrich (St. Louis, MO), respectively. Al(OH) 3 gel was prepared with 0.5 N NaOH and 0.5 N Al 2 (SO 4 ) 3 as previously described. 5) Animal Experiments Balb/c mice (6 weeks old, female, 18-20 g) were purchased from Japan SLC (Hamamatsu). The animals were housed in a temperature-controlled room 110 In the theory of traditional Chinese medicine (TCM), eqi ( ) circulates at the superficial portion of the body to guard against exopathogen. Gyokuheifusan (GHS; ), containing Astragalus Root, Atractylodes Rhizome, and Saposhnikovia Root, is a TCM formula to treat the insufficiency of eqi by invigorating qi and consolidating the superficial resistance. In this study, we evaluated the effect of GHS on murine antibody production against ovalbumin (OVA) used as exopathogen. Balb/c mice were sensitized with OVA and alum via intraperitoneal (i.p.) injection or intranasal (i.n.) infusion daily for 7 d. GHS was orally administered daily at the dose of 10-times amount of human daily dosage from 3 d before the sensitization for 14 d. Fourteen d after the final sensitization, the blood was collected, and the concentrations of OVA-specific or non-specific immunoglobulins were measured. When OVA was sensitized i.p., the concentration of OVA-specific IgG, IgG 1 , IgG 2a and IgA in the sera significantly increased by GHS-treatment. When OVA was sensitized i.n., GHS significantly reduce the concentration of OVA-specific IgG and IgG 1 in the sera. Non-specific immunoglobulins were not changed by GHS-treatment. It is suggested that GHS could stimulate immune responses when antigen had already been invaded into the inside of the body, and that GHS might consolidate the resistance of nasal mucosa to protect from the invasion of OVA, then OVA-specific antibodies in sera might be hypocritically suppressed. The present study might provide the experimental evidence for TCM theory.