Original Article
 

Downregulation of Aquaporin3 in Systemic Sclerosis Dermal Fibroblasts

Abstract

Skin dryness and thickening are hallmarks of systemic sclerosis (SSc) disease. Aquaporins (AQPs) are plasma membrane proteins that transport glycerol and water, resulting in water retention and skin hydration. Expression of AQPs has been evaluated in human normal skin. However, expression of these proteins in SSc dermal fibroblasts has not yet been reported. The aim of this study was to assess the expression profile of AQPs in dermal fibroblasts of SSc patients. Fibroblast cells were extracted from SSc and healthy skin biopsies and characterized using fibroblast surface protein antibody. The SYBR Green Real-time PCR was used to evaluate the mRNA expression of AQP1, 3, 5, 7, 9, and 10 in dermal fibroblasts. Immunoblotting was performed to confirm the results of Real-time PCR. Our data demonstrated that only AQP1, AQP3, and AQP9 were expressed in human skin fibroblasts. Moreover, the expression of AQP3 mRNA and protein were significantly decreased in SSc dermal fibroblasts compare to healthy fibroblasts. AQP3, which involves in skin hydration and wound healing through water and glycerol transmission, is downregulated in SSc fibroblasts. Based on previous studies and our results, it seems that SSc manifestations like skin dryness, abnormal wound healing, and fibrotic lesions may be related to downregulation of AQP3 in SSc fibroblasts. Therefore, induction of AQP3 expression can be a potential treatment to relieve SSc skin thickness in the future.

1. Varga J, Abraham D. Systemic sclerosis: a prototypic multisystem fibrotic disorder. J Clin Invest 2007; 117(3):557-67.

2. Balbir-Gurman A, Braun-Moscovici Y. Scleroderma - new aspects in pathogenesis and treatment. Best Pract Res Clin Rheumatol 2012; 26(1):13-24.

3. Korn JH. Immunologic aspects of scleroderma. Curr Opin Rheumatol 1989; 1(4):479-84.

4. Mauch C, Kreig T. Fibroblast-matrix interactions and their role in the pathogenesis of fibrosis. Rheum Dis Clin North Am; 16(1):93-107.

5. Abraham DJ, Krieg T, Distler J, Distler O. Overview of pathogenesis of systemic sclerosis. Rheumatology (Oxford) 2009; (48 Suppl 3):iii3-7.

6. Verkman AS, Mitra AK. Structure and function of aquaporin water channels. Am J Physiol Renal Physiol 2000; 278(1):F13-28.

7. Hara-Chikuma M, Verkman AS. Aquaporin-3 functions as a glycerol transporter in mammalian skin. Biol Cell 2005; 97(7):479-86.

8. Verkman AS. More than just water channels: unexpected cellular roles of aquaporins. J Cell Sci 2005; 118(Pt 15):3225-32.

9. Brown D, Katsura T, Kawashima M, Verkman AS, Sabolic I. Cellular distribution of the aquaporins: a family of water channel proteins. Histochem Cell Biol 1995; 104(1):1-9.

10. Verkman AS. Aquaporins at a glance. J Cell Sci 2011; 124(Pt 13):2107-12.

11. Verkman AS. Aquaporins: translating bench research to human disease. J Exp Biol 2009; 212(Pt 11):1707-15.

12. Hara-Chikuma M, Verkman AS. Roles of aquaporin-3 in the epidermis. J Invest Dermatol 2008; 128(9):2145-51.

13. Cao C, Sun Y, Healey S, Bi Z, Hu G, Wan S, et al. EGFR-mediated expression of aquaporin-3 is involved in human skin fibroblast migration. Biochem J 2006; 400(2):225-34.

14. Jinnin M. Mechanisms of skin fibrosis in systemic sclerosis. J Dermatol 2010; 37(1):11-25.

15. LeRoy EC. Increased collagen synthesis by scleroderma skin fibroblasts in vitro: a possible defect in the regulation or activation of the scleroderma fibroblast. J Clin Invest 1974; 54(4):880-9.

16. Moinzadeh P, Denton CP, Abraham D, Ong V, Hunzelmann N, Eckes B, et al. Biomarkers for skin involvement and fibrotic activity in scleroderma. J Eur Acad Dermatol Venereol 2012; 26(3):267-76.

17. Fluhr JW, Mao-Qiang M, Brown BE, Wertz PW, Crumrine D, Sundberg JP, et al. Glycerol regulates stratum corneum hydration in sebaceous gland deficient (asebia) mice. J Invest Dermatol 2003; 120(5):728-37.

18. Muratore M, Quarta L, Raho L, Costanza D, Frisenda S, Calcagnile F, et al. Management of cutaneous discomfort in patients with scleroderma: a clinical trial. Reumatismo 2013; 65(5):240-7.

19. Boury-Jamot M, Sougrat R, Tailhardat M, Le Varlet B, Bonte F, Dumas M, et al. Expression and function of aquaporins in human skin: Is aquaporin-3 just a glycerol transporter? Biochim Biophys Acta 2006; 1758(8):1034-42.

20. LeRoy EC, Black C, Fleischmajer R, Jablonska S, Krieg T, Medsger TA, Jr, et al. Scleroderma (systemic sclerosis): classification, subsets and pathogenesis. J Rheumatol 1988; 15(2):202-5.

21. Takashima A. Establishment of fibroblast cultures. Curr Protoc Cell Biol 2001; Chapter 2:Unit 2 1.

22. Loder RM, Martin GM. Establishment of human skin fibroblast cultures with an automatic wedgeinstrument. J Invest Dermatol 1966; 47(6):603-5.

23. Schmittgen TD, Livak KJ. Analyzing real-time PCR data by the comparative C(T) method. Nat Protoc 2008; 3(6):1101-8.

24. Bou-Gharios G, Osman J, Black C, Olsen I. Excess matrix accumulation in scleroderma is caused partly by differential regulation of stromelysin and TIMP-1 synthesis. Clin Chim Acta 1994; 231(1):69-78.

25. Hara-Chikuma M, Verkman AS. Aquaporin-3 facilitates epidermal cell migration and proliferation during wound healing. J Mol Med (Berl) 2008; 86(2):221-31.

26. Matthay MA, Thiery JP, Lafont F, Stampfer F, Boyer B. Transient effect of epidermal growth factor on the motility of an immortalized mammary epithelial cell line. J Cell Sci 1993; 106 ( Pt 3):869-78.

27. Fujii K, Dousaka-Nakajima N, Imamura S. Epidermal growth factor enhancement of HSC-1 human cutaneous squamous carcinoma cell adhesion and migration on type I collagen involves selective up-regulation of alpha 2 beta 1 integrin expression. Exp Cell Res 1995; 216(1):261-72.

28. Chen JD, Kim JP, Zhang K, Sarret Y, Wynn KC, Kramer RH, et al. Epidermal growth factor (EGF) promotes human keratinocyte locomotion on collagen by increasing the alpha 2 integrin subunit. Exp Cell Res 1993; 209(2):216-23.

29. Blay J, Brown KD. Epidermal growth factor promotes the chemotactic migration of cultured rat intestinal epithelial cells. J Cell Physiol 1985; 124(1):107-12.

30. Xu H, Xu Y, Zhang W, Shen L, Yang L, Xu Z. Aquaporin-3 positively regulates matrix metalloproteinases via PI3K/AKT signal pathway in human gastric carcinoma SGC7901 cells. J Exp Clin Cancer Res 2011; 30(1):86.

Files
IssueVol 16, No 3 (2017) QRcode
SectionOriginal Article(s)
Keywords
Aquaporin Fibroblast Gene expression Systemic sclerosis

Rights and permissions
Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
How to Cite
1.
Yousefi B, Mahmoudi M, Sarafnejad A, Karimizadeh E, Farhadi E, Jamshidi AR, Kavosi H, Aslani S, Gharibdoost F. Downregulation of Aquaporin3 in Systemic Sclerosis Dermal Fibroblasts. Iran J Allergy Asthma Immunol. 2017;16(3):228-234.