title: Regulatory effect of traditional Chinese medicinal formula Zuo-Gui-Wan on the Th17/Treg paradigm in mice with bone loss induced by estrogen deficiency.
abstract: BACKGROUND
  Bone loss is a common pathological condition induced by estrogen deficiency. The Th17/Treg paradigm, which can be skewed by estrogen, plays an important role in regulating bone metabolism
  
  
  OBJECTIVE
  The purpose of this study was to determine the role of the Th17/Treg shift in estrogen deficiency-induced bone loss in mouse models and to elucidate the immunopharmacologic mechanism of Zuo-Gui-Wan (ZGW) in preventing bone loss in this process by regulating Th17/Treg paradigm.
  
  
  METHODS
  Splenocytes of ovariectomized (Ovx) mice and naturally aged mice were isolated and Flow cytometry was used to detect the Th17/Treg subsets. In addition, serum estrodiol (E2) and serum C-terminal telopeptides of type Ι collagen (CTx) were detected by ELISA assay. Bone mineral density (BMD) of the left tibiae was measured by dual-energy X-ray absorptiometry. Moreover, Ovx mice were administrated with different doses of ZGW for 12 weeks, and BMD and Th17/Treg subsets were determined. Bone histomorphometry was observed by Hematoxylin and eosin (H&E) staining and serum protein levels of IL-6 were analyzed by ELISA assay. In addition, the mRNA and protein expression of RORγt and Foxp3 were detected by RT-PCR and Western blot respectively.
  
  
  RESULTS
  The Th17/Treg paradigm shifted to Th17 in estrogen-deficient mice both in the Ovx mice and the naturally aged mice. BMD and E2 levels negatively correlated with the Th17/Treg ratio. After ZGW administration, the BMD was enhanced markedly in the Ovx mice as well as in the naturally aged mice. Both the mRNA and protein expressions of IL-6 and RORγt were decreased, whereas those of Foxp3 were increased significantly after ZGW administration.
  
  
  CONCLUSIONS
  Th17/Treg shift involved in the bone loss induced by estrogen deficiency. ZGW prevented bone loss efficiently and skewed Th17/Treg paradigm towards Treg without enhancing E2.
authors.affiliation: Key Laboratory for Tumor Immunology and Traditional Chinese Medicine Immunology, Institute of Basic Medicine, Shandong Academy of Medical Sciences, 18877 Jingshi Road, Jinan 250062, China; Key Laboratory for Rare & Uncommon Disease, Institute of Basic Medicine, Shandong Academy of Medical Sciences, 18877 Jingshi Road, Jinan 250062, China; Key Laboratory of Biotechnology Drugs of the Ministry of Public Health, Institute of Basic Medicine, Shandong Academy of Medical Sciences, 18877 Jingshi Road, Jinan 250062, China; Laboratory for Immunopharmacology of State Administration of Traditional Chinese Medicine, Institute of Basic Medicine, Shandong Academy of Medical Sciences, 18877 Jingshi Road, Jinan 250062, China.
authors.forename: Nannan
authors.initials: N
authors.lastname: Lai
authors.affiliation: Department of reproductive center, Yantai Yuhuangding Hospital, Shandong University, 44 Wenhua xi Road, Jinan 250012, China.
authors.forename: Zhirong
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: Affiliated Hospital of Shandong University of Traditional Chinese Medicine, 42 Wenhua xi Road, Jinan 250011, China.
authors.forename: Bin
authors.initials: B
authors.lastname: Wang
authors.affiliation: Affiliated Hospital of Shandong University of Traditional Chinese Medicine, 42 Wenhua xi Road, Jinan 250011, China.
authors.forename: Xiuming
authors.initials: X
authors.lastname: Miao
authors.affiliation: Key Laboratory for Tumor Immunology and Traditional Chinese Medicine Immunology, Institute of Basic Medicine, Shandong Academy of Medical Sciences, 18877 Jingshi Road, Jinan 250062, China; Key Laboratory for Rare & Uncommon Disease, Institute of Basic Medicine, Shandong Academy of Medical Sciences, 18877 Jingshi Road, Jinan 250062, China; Key Laboratory of Biotechnology Drugs of the Ministry of Public Health, Institute of Basic Medicine, Shandong Academy of Medical Sciences, 18877 Jingshi Road, Jinan 250062, China; Laboratory for Immunopharmacology of State Administration of Traditional Chinese Medicine, Institute of Basic Medicine, Shandong Academy of Medical Sciences, 18877 Jingshi Road, Jinan 250062, China.
authors.forename: Yuqi
authors.initials: Y
authors.lastname: Guo
authors.affiliation: Key Laboratory for Tumor Immunology and Traditional Chinese Medicine Immunology, Institute of Basic Medicine, Shandong Academy of Medical Sciences, 18877 Jingshi Road, Jinan 250062, China; Key Laboratory for Rare & Uncommon Disease, Institute of Basic Medicine, Shandong Academy of Medical Sciences, 18877 Jingshi Road, Jinan 250062, China; Key Laboratory of Biotechnology Drugs of the Ministry of Public Health, Institute of Basic Medicine, Shandong Academy of Medical Sciences, 18877 Jingshi Road, Jinan 250062, China; Laboratory for Immunopharmacology of State Administration of Traditional Chinese Medicine, Institute of Basic Medicine, Shandong Academy of Medical Sciences, 18877 Jingshi Road, Jinan 250062, China.
authors.forename: Chengfang
authors.initials: C
authors.lastname: Yao
authors.affiliation: Key Laboratory for Tumor Immunology and Traditional Chinese Medicine Immunology, Institute of Basic Medicine, Shandong Academy of Medical Sciences, 18877 Jingshi Road, Jinan 250062, China; Key Laboratory for Rare & Uncommon Disease, Institute of Basic Medicine, Shandong Academy of Medical Sciences, 18877 Jingshi Road, Jinan 250062, China; Key Laboratory of Biotechnology Drugs of the Ministry of Public Health, Institute of Basic Medicine, Shandong Academy of Medical Sciences, 18877 Jingshi Road, Jinan 250062, China; Laboratory for Immunopharmacology of State Administration of Traditional Chinese Medicine, Institute of Basic Medicine, Shandong Academy of Medical Sciences, 18877 Jingshi Road, Jinan 250062, China.
authors.forename: Zhaoxia
authors.initials: Z
authors.lastname: Wang
authors.affiliation: Key Laboratory for Tumor Immunology and Traditional Chinese Medicine Immunology, Institute of Basic Medicine, Shandong Academy of Medical Sciences, 18877 Jingshi Road, Jinan 250062, China; Key Laboratory for Rare & Uncommon Disease, Institute of Basic Medicine, Shandong Academy of Medical Sciences, 18877 Jingshi Road, Jinan 250062, China; Key Laboratory of Biotechnology Drugs of the Ministry of Public Health, Institute of Basic Medicine, Shandong Academy of Medical Sciences, 18877 Jingshi Road, Jinan 250062, China; Laboratory for Immunopharmacology of State Administration of Traditional Chinese Medicine, Institute of Basic Medicine, Shandong Academy of Medical Sciences, 18877 Jingshi Road, Jinan 250062, China.
authors.forename: Li
authors.initials: L
authors.lastname: Wang
authors.affiliation: Qianfo Hospital of Shandong Province, 16766 Jingshi Road, Jinan 250014, China.
authors.forename: Ruiping
authors.initials: R
authors.lastname: Ma
authors.affiliation: Key Laboratory for Tumor Immunology and Traditional Chinese Medicine Immunology, Institute of Basic Medicine, Shandong Academy of Medical Sciences, 18877 Jingshi Road, Jinan 250062, China; Key Laboratory for Rare & Uncommon Disease, Institute of Basic Medicine, Shandong Academy of Medical Sciences, 18877 Jingshi Road, Jinan 250062, China; Key Laboratory of Biotechnology Drugs of the Ministry of Public Health, Institute of Basic Medicine, Shandong Academy of Medical Sciences, 18877 Jingshi Road, Jinan 250062, China. Electronic address: 786735868@qq.com.
authors.forename: Xia
authors.initials: X
authors.lastname: Li
authors.affiliation: Key Laboratory for Tumor Immunology and Traditional Chinese Medicine Immunology, Institute of Basic Medicine, Shandong Academy of Medical Sciences, 18877 Jingshi Road, Jinan 250062, China; Key Laboratory for Rare & Uncommon Disease, Institute of Basic Medicine, Shandong Academy of Medical Sciences, 18877 Jingshi Road, Jinan 250062, China; Key Laboratory of Biotechnology Drugs of the Ministry of Public Health, Institute of Basic Medicine, Shandong Academy of Medical Sciences, 18877 Jingshi Road, Jinan 250062, China. Electronic address: jiangguosh@163.com.
authors.forename: Guosheng
authors.initials: G
authors.lastname: Jiang
mesh_terms: D000818:Animals; D015519:Bone Density / Q000187:drug effects; D024042:Collagen Type I / Q000378:metabolism; D004365:Drugs, Chinese Herbal / Q000494:pharmacology*; D004967:Estrogens / Q000172:deficiency*; D005260:Female; D051858:Forkhead Transcription Factors / Q000378:metabolism; D015850:Interleukin-6 / Q000378:metabolism; D008516:Medicine, Chinese Traditional / Q000379:methods; D051379:Mice; D008807:Mice, Inbred BALB C; D057132:Nuclear Receptor Subfamily 1, Group F, Member 3 / Q000378:metabolism; D010024:Osteoporosis / Q000188:drug therapy* / Q000378:metabolism; D010052:Ovariectomy / Q000379:methods; D010455:Peptides / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D050378:T-Lymphocytes, Regulatory / Q000187:drug effects* / Q000378:metabolism; D058504:Th17 Cells / Q000187:drug effects* / Q000378:metabolism
chemical_list: D024042:Collagen Type I; D004365:Drugs, Chinese Herbal; D004967:Estrogens; D051858:Forkhead Transcription Factors; C498833:Foxp3 protein, mouse; D015850:Interleukin-6; D057132:Nuclear Receptor Subfamily 1, Group F, Member 3; D010455:Peptides; D012333:RNA, Messenger; C052929:collagen type I trimeric cross-linked peptide; C542063:zuo gui wan
keywords: Bone loss; Cytokine; Estrogen deficiency; Th17/Treg paradigm; Transcription factor; Zuo-Gui-Wan