title: Evaluation of safety of modified-Danggui Buxue Tang in rodents:immunological, toxicity and hormonal aspects.
abstract: BACKGROUND
  Radix Astragali (RA), Radix Angelicae Sinensis (RAS) and Folium Epimedii (FE) are three of the extensively applied herbs among traditional Chinese medicines for gynecological disorders and osteoporosis. A derivative herbal formula-RRF, consisting of the three medicines with a weight ratio of 5:1:5, is derived from a famous Chinese herbal formula-Danggui Buxue Tang (DBT). RRF has shown noteworthy perimenopause ameliorating effect in both ovariectomized rats and natural aging female rats, which might represent a promising candidate for the treatment of perimenopausal disorders. The aim of this study was to evaluate its immunological potential, chronic toxicity and reproductive effects by 26-week repeated daily administration in female rats, in order to optimize its safe use.
  
  
  METHODS
  The effect of RRF on immunological function was studied by macrophage phagocytosis, immune organ index, serum immunoglobulin level as well as delayed type hypersensitivity (DTH) in mice. For toxicity assessment, acute toxicity study was performed according to fixed dose procedure with a single oral administration of RRF to mice. In the oral chronic toxicity, 120 female rats were administrated RRF orally in 0, 1100, 4400, or 8800mg/kg/day doses for 26 weeks. Clinical signs, mortality, body weights, feed consumption, haemato-biochemical parameters, organ weights, histopathology and reproductive hormone profiles were examined at the end of the 13- and 26-week dosing period, as well as after the 4-week recovery period.
  
  
  RESULTS
  Oral administration of RRF at three doses (282, 564 and 1128mg/kg) significantly increased the indices of phagocytosis K, as compared with prednisone acetate (PR) group (p<0.05 or 0.01). Exposure of RRF dose-dependently boosted circulating serum IgM level (all p<0.01) in response to CRBC in PR-induced mice. Furthermore, RRF treatment elicited a significant increment (all p<0.01) in DNFB-induced DTH response and the immune organ indices in a dose-dependent manner in mice, in parellel to DNFB-induced group. In the single dose acute toxicity and repeated dose 90-day chronic toxicity investigations, no toxic signs/mortality were observed. RRF treatment did not cause any toxicologically significant changes in clinical signs, food consumption, body weight, relative organ weight, hematological parameters, clinical chemistry, gross pathology and histopathology between treatment and control groups. No treatment related gross/histopathological lesions were observed and no target organ was identified. Long-term repeated administration of RRF exerted a significant promotion on serum level of steroid hormone estradiol, progesterone and testosterone release, along with decrease of circulating pituitary follicle stimulating hormone, luteinizing hormone, and prolactin levels in female rats. The No Observed Adverse Effect Level (NOAEL) of RRF was determined to be over 8800mg/kg/day for elderly female rats, a dose that was equivalent to 50 times of human dose.
  
  
  CONCLUSIONS
  The present investigation demonstrated that RRF possessed appreciable immunopotentiating activity and had a relatively wide margin of safety. Long-term treatment of RRF exhibited estrogenic properties, and retarded certain age-associated degenerations. RRF might have the potential for further development as a safe and effective alternative/complementary to conventional medication in relieving perimenopausal symptoms.
authors.affiliation: Guangdong Provincial Key Laboratory of Clinical Research on Traditional Chinese Medicine Syndrome, The Second Affiliated Hospital, Guangzhou University of Chinese Medicine, Guangzhou 510120, PR China.
authors.forename: Jian-Hui
authors.initials: JH
authors.lastname: Xie
authors.affiliation: Dongguan Mathematical Engineering Academy of Chinese Medicine, Guangzhou University of Chinese Medicine, Dongguan 523000, PR China; The Second Affiliated Hospital, Guangzhou University of Chinese Medicine, Guangzhou, 510120, PR China.
authors.forename: Zhi-Wei
authors.initials: ZW
authors.lastname: Chen
authors.affiliation: Department of Orthopaedics, Dongguan Kanghua Hospital, Dongguan 523080, PR China.
authors.forename: Ya-Wei
authors.initials: YW
authors.lastname: Pan
authors.affiliation: Department of Orthopaedics, Dongguan Kanghua Hospital, Dongguan 523080, PR China.
authors.forename: De-Min
authors.initials: DM
authors.lastname: Luo
authors.affiliation: School of Chinese Materia Medica, Guangzhou University of Chinese Medicine, Guangzhou, 510006, PR China.
authors.forename: Zi-Ren
authors.initials: ZR
authors.lastname: Su
authors.affiliation: The Second Affiliated Hospital, Guangzhou University of Chinese Medicine, Guangzhou, 510120, PR China.
authors.forename: Hai-Ming
authors.initials: HM
authors.lastname: Chen
authors.affiliation: Department of Pharmacology, Guiyang Medical University, Guiyang 550025, PR China.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Qin
authors.affiliation: School of Continuing Education, Guangzhou University of Chinese Medicine, Guangzhou 510006, PR China. Electronic address: hsq@gzucm.edu.cn.
authors.forename: Shui-Qing
authors.initials: SQ
authors.lastname: Huang
authors.affiliation: Department of Orthopaedics, Dongguan Kanghua Hospital, Dongguan 523080, PR China. Electronic address: lei.chenggaoo@163.com.
authors.forename: Gao
authors.initials: G
authors.lastname: Lei
mesh_terms: D000284:Administration, Oral; D000818:Animals; D027883:Astragalus Plant / Q000009:adverse effects / Q000737:chemistry; D004365:Drugs, Chinese Herbal / Q000009:adverse effects* / Q000494:pharmacology*; D005260:Female; D006728:Hormones / Q000378:metabolism*; D008516:Medicine, Chinese Traditional / Q000009:adverse effects; D051379:Mice; D019055:No-Observed-Adverse-Effect Level; D047648:Perimenopause / Q000187:drug effects; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D004365:Drugs, Chinese Herbal; D006728:Hormones; C477279:danggui buxue decoction
keywords: Chronic toxicity; Danggui Buxue Tang; Immunological; Modified herbal formula; Reproductive effect