Table 1.
Summary of the effects of ROCK 1 and ROCK 2 on the physical properties of 3D human corneal stroma fibroblasts (HCSFs) spheroids and gene expression of 2D and 3D cultured HCSFs.
| 1 mM Rip | 10 mM Rip | 1 mM KD025 | 10 mM KD025 | |
|---|---|---|---|---|
| Size | ↑↑↑↑ | ↑↑↑↑ | (–) | ↑↑ |
| Stiffness | (–) | ↓↓ | (–) | ↑ |
| COL 1 | ||||
| 2D | (–) | (–) | (–) | ↓ |
| 3D | ↓↓↓↓ | ↓↓↓↓ | ↓ | ↓↓↓↓ |
| COL 4 | ||||
| 2D | (–) | ↓ | (–) | ↑↑ |
| 3D | (–) | ↓ | (–) | (–) |
| COL 6 | ||||
| 2D | (–) | (–) | (–) | (–) |
| 3D | (–) | (–) | ↑↑↑ | (–) |
| FN | ||||
| 2D | (–) | ↓↓↓↓ | (–) | (–) |
| 3D | ↓↓ | ↓↓↓↓ | ↓↓ | ↓↓↓↓ |
| TIMP1 | ||||
| 2D | (–) | (–) | (–) | (–) |
| 3D | ↓↓ | ↓↓↓↓ | ↓↓↓↓ | (–) |
| TIMP2 | ||||
| 2D | (–) | (–) | (–) | ↓↓↓↓ |
| 3D | ↓↓↓ | ↓↓↓↓ | (–) | (–) |
| TIMP3 | ||||
| 2D | (–) | ↑↑ | (–) | ↓↓ |
| 3D | (–) | ↓↓↓↓ | (–) | ↓↓↓↓ |
| TIMP4 | ||||
| 2D | ↓↓ | (–) | ↑↑↑↑ | (–) |
| 3D | (–) | ↑↑↑↑ | (–) | (–) |
| MMP2 | ||||
| 2D | (–) | (–) | (–) | ↓↓↓↓ |
| 3D | ↓ | (–) | (–) | (–) |
| MMP3 | ||||
| 2D | (–) | (–) | ↑↑↑↑ | ↑↑↑ |
| 3D | (––) | ↑↑↑↑ | (–) | (–) |
| MMP9 | ||||
| 2D | (–) | (–) | (–) | (–) |
| 3D | (–) | (–) | ↓↓↓↓ | ↓↓ |
| MMP14 | ||||
| 2D | (–) | (–) | (–) | ↓↓↓↓ |
| 3D | (–) | ↑↑↑↑ | (–) | (–) |
| GRP 78 | ||||
| 2D | (–) | (–) | (–) | (–) |
| 3D | (–) | (–) | ↓ | ↓↓↓↓ |
| GRP 94 | ||||
| 2D | (–) | ↓↓ | ↓↓↓ | ↓↓↓↓ |
| 3D | ↑ | (–) | ↓↓↓↓ | ↓↓↓↓ |
| IRE 1 | ||||
| 2D | (–) | (–) | (–) | ↓ |
| 3D | (–) | (–) | (–) | (–) |
| ATF 6 | ||||
| 2D | (–) | (–) | (–) | ↓↓ |
| 3D | (–) | (–) | ↓↓↓ | ↓↓↓↓ |
| PERK | ||||
| 2D | (–) | (–) | (–) | ↓↓ |
| 3D | (–) | (–) | ↓↓↓↓ | ↓↓↓↓ |
| ATF 4 | ||||
| 2D | (–) | ↓↓↓↓ | ↓↓↓↓ | ↓↓↓↓ |
| 3D | (–) | (–) | ↓↓↓↓ | ↓↓↓↓ |
| XBP 1 | ||||
| 2D | (–) | (–) | (–) | ↓↓ |
| 3D | (–) | ↓ | (–) | ↓↓↓↓ |
| sXBP 1 | ||||
| 2D | (–) | (–) | (–) | ↑ |
| 3D | ↓↓ | ↑↑ | ↓↓↓↓ | ↓↓↓↓ |
| CHOP | ||||
| 2D | (–) | (–) | (–) | ↓↓↓↓ |
| 3D | (–) | ↓ | ↓↓ | ↓↓↓↓ |
Rip: ripasudil, 2D: two-dimension culture, 3D: three-dimension culture, COL: collagen, FN: fibronectin, TIMP: tissue inhibitor of metalloproteinase, MMP: matrix metalloproteinase, protein kinase RNA-like endoplasmic reticulum kinase (PERK), activating transcription factor 6 (ATF6) and the inositol-requiring enzyme 1 (IRE1), and their downstream factors including the glucose regulator protein (GRP)78, GRP94, the X-box binding protein-1 (XBP1), spliced XBP1 (sXBP1) and CCAAT/enhancer-binding protein homologous protein (CHOP), (–): no significant change, ↑: significant increase (p < 0.05), ↑↑: significant increase (p < 0.01), ↑↑↑: significant increase (p < 0.005), ↓: significant decrease (p < 0.05), ↓↓: significant decrease (p < 0.01), ↓↓↓: significant decrease (p < 0.005).