Involvement of P38MAPK in human corneal endothelial cell migration induced by TGF-β2

被引：41

作者：

Joko, Takeshi ^{[1
]}

Shiraishi, Atsushi ^{[1
,2
]}

Akune, Yoko ^{[1
]}

Tokumaru, Sho ^{[3
]}

Kobayashi, Takeshi ^{[2
,4
]}

Miyata, Kazunori ^{[6
]}

Ohashi, Yuichi ^{[1
,5
]}

机构：

[1] Ehime Univ, Dept Ophthalmol, Grad Sch Med, Toon, Ehime 7910295, Japan

[2] Ehime Univ, Dept Stem Cell Biol, Grad Sch Med, Toon, Ehime 7910295, Japan

[3] Ehime Univ, Dept Dermatol, Grad Sch Med, Toon, Ehime 7910295, Japan

[4] Ehime Univ, Dept Ophthalmol & Regenerat Med, Grad Sch Med, Toon, Ehime 7910295, Japan

[5] Ehime Univ, Dept Infect Dis, Grad Sch Med, Toon, Ehime 7910295, Japan

[6] Miyata Eye Hosp, Miyakonojo, Japan

来源：

EXPERIMENTAL EYE RESEARCH | 2013年 / 108卷

关键词：

P38MAPK; TGF-beta(2); corneal endothelial wound healing; cell migration; FGF-2; GROWTH-FACTOR-BETA; HUMAN AQUEOUS-HUMOR; IN-VITRO; TGF-BETA; EPITHELIAL-CELLS; MAP KINASE; PROLIFERATION; P38; EXPRESSION; P27(KIP1);

D O I：

10.1016/j.exer.2012.11.018

中图分类号：

R77 [眼科学];

学科分类号：

100212 ;

摘要：

Because human corneal endothelial cells do not proliferate once the endothelial monolayer is formed, corneal wound healing is thought to be mediated by cell enlargement or migration rather than proliferation. However, the cellular mechanisms involved in corneal wound healing have not been fully determined. Because transforming growth factor-beta(2) (TGF-beta(2)) isoform is present in high concentrations in normal human aqueous humor, it may play a role in human corneal endothelial cell wound healing. The purpose of this study was to determine the effect of TGF-beta(2) on the proliferation and migration of cultured human corneal endothelial cells (HCECs). To achieve this, we first examined the effect of TGF-beta(2) on the wound closure rate in an in vitro HCEC wound healing model. However, unexpectedly TGF-beta(2) had no effect on the wound closure rate in this model. Therefore, a real-time cell electronic sensing (RT-CES) system and the BrdU incorporation assay were used to determine the effect of TGF-beta(2) (0.1-10 ng/ml) on cultured HCEC proliferation during in vitro wound healing. The specificity of this effect was confirmed by adding the TGF-beta receptor I kinase inhibitor. TGF-beta(2) inhibited the proliferation of HCECs in a dose dependent way and was blocked by TGF-beta receptor I kinase inhibitor. Next, the Boyden chamber assay was used to determine how TGF-beta(2) (10 ng/ml) affect HCEC migration. Exposure to TGF-beta(2) increased cell migration, and a synergistic effect was observed when FGF-2 was added. To determine whether the mitogen-activated protein kinase (MAPK) signaling pathway is involved in the migration of HCECs, western blot analysis and Bio-Plex (TM) suspension array were used to detect phosphorylation of Erk1/2, p38, and JNK in HCECs stimulated by TGF-beta(2) and/or FGF-2. The effect of the p38 MAPK inhibitor, SB239063 (10 mu M), on TGF-beta(2) and/or FGF-2-induced cellular migration was determined by the Boyden chamber assay. Both TGF-beta(2) and FGF-2-induced p38 phosphorylation, and a synergistic effect was observed with exposure to both growth factors. SB 239063 inhibited TGF-beta 2 and FGF-2-induced migration of HCECs. These results indicate that TGF-beta(2) reduces proliferation but stimulates migration of cultured HCECs. In addition, TGF-beta(2) and FGF-2 may have synergistic effects on the migration of HCECs mediated by p38 MAPK phosphorylation. (c) 2012 Elsevier Ltd. All rights reserved.

引用

页码：23 / 32

页数：10

共 41 条

[21] Human Corneal Endothelial Cells Employ Phosphorylation of p27Kip1 at Both Ser10 and Thr187 Sites for FGF-2-Mediated Cell Proliferation via PI 3-Kinase [J].

Lee, Jeong Goo ;

Song, Jong-Suk ;

Smith, Ronald E. ;

Kay, EunDuck P. .

INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE, 2011, 52 (11) :8216-8223

[22]

LING TL, 1988, INVEST OPHTH VIS SCI, V29, P1407

[23] The logic of TGFβ signaling [J].

Massague, Joan ;

Gomis, Roger R. .

FEBS LETTERS, 2006, 580 (12) :2811-2820

[24] Effect of donor age on morphologic variation of cultured human corneal endothelial cells [J].

Miyata, K ;

Drake, J ;

Osakabe, Y ;

Hosokawa, Y ;

Hwang, D ;

Soya, K ;

Oshika, T ;

Amano, S .

CORNEA, 2001, 20 (01) :59-63

[25]

MURPHY C, 1984, INVEST OPHTH VIS SCI, V25, P312

[26] Transforming growth factor β2 levels in the aqueous humor in different types of glaucoma and the relation to filtering bleb development [J].

Picht, G ;

Welge-Luessen, U ;

Grehn, F ;

Lütjen-Drecoll, E .

GRAEFES ARCHIVE FOR CLINICAL AND EXPERIMENTAL OPHTHALMOLOGY, 2001, 239 (03) :199-207

[27] THE ROLE OF EXOGENOUS ENDOGENOUS BASIC FIBROBLAST GROWTH-FACTOR (FGF2) AND TRANSFORMING GROWTH-FACTOR-BETA (TGF-BETA-1) ON HUMAN CORNEAL ENDOTHELIAL-CELLS PROLIFERATION IN-VITRO [J].

RIECK, P ;

OLIVER, L ;

ENGELMANN, K ;

FUHRMANN, G ;

HARTMANN, C ;

COURTOIS, Y .

EXPERIMENTAL CELL RESEARCH, 1995, 220 (01) :36-46

[28] Intracellular signaling pathway of FGF-2-modulated corneal endothelial cell migration during wound healing in vitro [J].

Rieck, PW ;

Cholidis, S ;

Hartmann, C .

EXPERIMENTAL EYE RESEARCH, 2001, 73 (05) :639-650

[29] Role of p38 MAP kinase in regulation of cell migration and proliferation in healing corneal epithelium [J].

Saika, S ;

Okada, Y ;

Miyamoto, T ;

Yamanaka, O ;

Ohnishi, Y ;

Ooshima, A ;

Liu, CY ;

Weng, D ;

Kao, WWY .

INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE, 2004, 45 (01) :100-109

[30]

Saika Shizuya, 2004, Cornea, V23, pS25, DOI 10.1097/01.ico.0000136668.41000.73

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