title: 20S-hydroxyvitamin D3, noncalcemic product of CYP11A1 action on vitamin D3, exhibits potent antifibrogenic activity in vivo.
abstract: BACKGROUND
  There is no effective treatment for systemic sclerosis and related fibrosing diseases. Recently the action of CYP11A1 on vitamin D(3) was shown to produce biologically active 20S-hydroxyvitamin D [20(OH)D(3)] and 20,23(OH)(2)D(3), 20,22(OH)(2)D(3), and 17,20,23(OH)(3)D(3).
  
  
  OBJECTIVE
  Because 20(OH)D(3) is noncalcemic (nontoxic) in vivo at very high doses, we evaluated its antifibrogenic activities both in vitro and in vivo. Because it is further metabolized by CYP11A1, we also tested preclinical utilities of its hydroxyderivatives, especially 20,23(OH)(2)D(3).
  
  
  METHODS
  Human dermal fibroblasts from scleroderma and normal donors were used to test the efficiency of hydroxyvitamin D derivatives in inhibiting TGF-β1-induced collagen and hyaluronan synthesis and inhibiting cell proliferation. The in vivo activity of 20(OH)D(3) was tested using bleomycin-induced sclerosis in C57BL/6 mice.
  
  
  RESULTS
  20(OH)D(3) and 20,23(OH)(2)D(3) inhibited TGF-β1-induced collagen and hyaluronan synthesis similarly to 1,25(OH)(2)D(3) in cultured human fibroblasts. Also, 20(OH)D(3), 20,23(OH)(2)D(3), and 1,25(OH)(2)D(3) suppressed TGF-β1-induced expression of COL1A2, COL3A1, and hyaluronan synthase-2 mRNA, indicating that they regulate these matrix components at the transcriptional level. 20(OH)D(3), 20,23(OH)(2)D(3), 20,22(OH)(2)D(3), and 17,20,23(OH)(3)D(3) inhibited proliferation of dermal fibroblasts with comparable potency with 1,25(OH)(2)D(3), with 20(OH)D(2) being less active and 1α(OH)D(3) being almost inactive. 20,23(OH)(2)D(3) at 3 μg/kg had no effect on serum Ca(++) or fibroblast growth factor-23 levels and did not cause any noticeable signs of morbidity. 20(OH)D(3) markedly suppressed fibrogenesis in mice given sc bleomycin as demonstrated by total collagen content and hematoxylin and eosin staining of skin biopsies.
  
  
  CONCLUSIONS
  20(OH)D(3) is an excellent candidate for preclinical studies on scleroderma, with other CYP11A1-derived products of its metabolism deserving further testing for antibrogenic activity.
authors.affiliation: Department of Pathology, University of Tennessee Health Science Center, Memphis, Tennessee 38163, USA.
authors.forename: Andrzej
authors.initials: A
authors.lastname: Slominski
authors.forename: Zorica
authors.initials: Z
authors.lastname: Janjetovic
authors.forename: Robert C
authors.initials: RC
authors.lastname: Tuckey
authors.forename: Minh N
authors.initials: MN
authors.lastname: Nguyen
authors.forename: Keka G
authors.initials: KG
authors.lastname: Bhattacharya
authors.forename: Jin
authors.initials: J
authors.lastname: Wang
authors.forename: Wei
authors.initials: W
authors.lastname: Li
authors.forename: Yan
authors.initials: Y
authors.lastname: Jiao
authors.forename: Weikuan
authors.initials: W
authors.lastname: Gu
authors.forename: Monica
authors.initials: M
authors.lastname: Brown
authors.forename: Arnold E
authors.initials: AE
authors.lastname: Postlethwaite
mesh_terms: D000818:Animals; D002112:Calcifediol / Q000031:analogs & derivatives* / Q000494:pharmacology; D049109:Cell Proliferation / Q000187:drug effects*; D002478:Cells, Cultured; D003094:Collagen / Q000096:biosynthesis; D004195:Disease Models, Animal; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D006820:Hyaluronic Acid / Q000096:biosynthesis; D051379:Mice; D008810:Mice, Inbred C57BL; D012595:Scleroderma, Systemic / Q000378:metabolism*; D012867:Skin / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism
chemical_list: C480634:20-hydroxyvitamin D3; D006820:Hyaluronic Acid; D003094:Collagen; D002112:Calcifediol