Inhibition by female sex hormones of collagen gel contraction mediated by retinal pigment epithelial cells

Invest Ophthalmol Vis Sci. 2014 Apr 21;55(4):2621-30. doi: 10.1167/iovs.13-13501.

Abstract

Purpose: Collagen contraction mediated by retinal pigment epithelial (RPE) cells contributes to the pathogenesis of proliferative vitreoretinopathy (PVR). We examined the effects of sex hormones on this process.

Methods: Mouse RPE cells were cultured in a type I collagen gel and exposed to 17β-estradiol, progesterone, or dehydro-epiandrosterone. Collagen contraction induced by transforming growth factor-β2 (TGF-β2) was determined by measurement of gel diameter. Expression of α-smooth muscle actin (α-SMA), as well as phosphorylation of Smad2 and myosin light chain (MLC), was examined by immunoblot analysis. Matrix metalloproteinase (MMP) release was evaluated by gelatin zymography. Fibronectin and interleukin-6 secretion was measured with immunoassays.

Results: The female sex hormones 17β-estradiol and progesterone inhibited TGF-β2-induced collagen contraction mediated by RPE cells, whereas the male sex hormone dehydro-epiandrosterone had no such effect. The TGF-β2-induced release of MMP-2 and MMP-9 from RPE cells was also inhibited by 17β-estradiol and progesterone, and the MMP inhibitor GM6001 attenuated TGF-β2-induced collagen contraction. Expression of the mesenchymal markers α-SMA and fibronectin, interleukin-6 release, and Smad2 and MLC phosphorylation induced by TGF-β2 were all inhibited by 17β-estradiol and progesterone. Immunohistochemical analysis also detected nuclear immunoreactivity for estrogen and progesterone receptors in proliferative fibrocellular membranes of PVR patients.

Conclusions: Female sex hormones inhibited TGF-β2-induced collagen contraction mediated by RPE cells. This action appeared to be mediated through inhibition both of MMP, α-SMA, and fibronectin expression as well as of Smad2 and MLC phosphorylation. Female sex hormones might thus prove effective for the treatment of PVR.

Keywords: ECM; TGF-β; collagen; retinal pigment epithelium.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Aged
  • Aged, 80 and over
  • Animals
  • Blotting, Western
  • Cells, Cultured
  • Collagen / drug effects*
  • Collagen / metabolism
  • Disease Models, Animal
  • Epithelial-Mesenchymal Transition / drug effects
  • Female
  • Gels
  • Gonadal Steroid Hormones / pharmacology*
  • Humans
  • Immunohistochemistry
  • Male
  • Mice
  • Microscopy, Fluorescence
  • Middle Aged
  • Retinal Pigment Epithelium / cytology
  • Retinal Pigment Epithelium / drug effects
  • Retinal Pigment Epithelium / metabolism*
  • Vitreoretinopathy, Proliferative / metabolism*
  • Vitreoretinopathy, Proliferative / pathology

Substances

  • Gels
  • Gonadal Steroid Hormones
  • Collagen