Combined effects of interleukin 1beta and mechanical stretching on Collagen I synthesis and Lumican expression in corneal fibroblasts

doi:10.3969/j.issn.2095-4344.0924

Abstract

Abstract:

BACKGROUND: Secondary keratoconus is one of the complications after corneal refractive surgery, but the mechanism remains unclear.
OBJECTIVE: To study the effects of interleukin-1β and mechanical stretching on the synthesis of the Collagen I and Lumican in corneal fibroblasts.
METHODS: Rabbit corneal fibroblasts were cultured in vitro and were treated by different concentrations of interleukin-1β and different amplitudes of mechanical stretching (5%, 10%, 15%) for 12, 24 or 36 hours. The expression of Collagen I and Lumican was detected by real-time fluorescent quantitative PCR.
RESULTS AND CONCLUSION: Interleukin-1β decreased the expression of Collagen I α1 at 12 hours and increased the expression of Lumican at 24 hours (P < 0.05). 5% stretching alone increased the expression of Collagen I α1 at 24 and 36 hours (P < 0.05), but 10% and 15% stretching alone decreased the expression of Collagen I α1 and Collagen I α2 at 12, 24 and 36 hours (P < 0.05). 5% stretching alone increased the expression of Lumican at 12 hours (P < 0.05), while 10% and 15% stretching alone decreased the expression of Lumican at 12 hours (P < 0.05). Moreover, 5%, 10% and 15% stretching alone increased the expression of Lumican at 24 and 36 hours (P < 0.05). Reduced expression of Collagen I α1 and Collagen I α2 and increased expression of Lumican in corneal fibroblasts were observed at 24 and 36 hours after combined treatment with interleukin-1β and mechanical stretching. These results reveal that interleukin-1β can inhibit the synthesis of Collagen I and promote the expression of Lumican. Low-amplitude stretching can increase collagen synthesis; however, middle-to-large- amplitude stretching can inhibit its expression in corneal fibroblasts. The synthesis of collagen can be suppressed and the expression of Lumican can be increased under the combined use of interleukin-1β and stretching.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Cornea, Fibroblasts, Interleukin-1beta, Collagen Type I, Tissue Engineering

CLC Number:

R394.2

Feng Peng-fei, Li Xiao-na, Rong Shuo, Chen Wei-yi, Wang Xiao-jun. Combined effects of interleukin 1beta and mechanical stretching on Collagen I synthesis and Lumican expression in corneal fibroblasts[J]. Chinese Journal of Tissue Engineering Research, 2018, 22(25): 4077-4082.

Figures/Tables 1

2.1 白细胞介素1β与机械牵拉对角膜成纤维细胞CollagenⅠα1的影响白细胞介素1β与机械牵拉共同作用于角膜成纤维细胞，实时荧光定量PCR检测CollagenⅠα1 mRNA的表达，见图1。未添加白细胞介素1β时，与静态对照组相比，5%，10%，15%分别牵拉12 h使CollagenⅠα1表达下降，差异有显著性意义(P < 0.05)；牵拉24 h，CollagenⅠα1的表达在5%牵拉时表达升高，在10%，15%牵拉时表达下降，差异有显著性意义(P < 0.05)；牵拉36 h，CollagenⅠα1的表达在5%牵拉时表达升高，在10%，15%牵拉时表达下降，差异有显著性意义(P < 0.05)。在白细胞介素1β为0.2 μg/L和0.4 μg/L时，与静态对照组相比，CollagenⅠα1的表达在5%和10%牵拉12 h时没有变化，而在15%牵拉12 h时降低，差异有显著性意义(P < 0.05)；与静态对照组相比，CollagenⅠα1的表达在5%，10%，15%牵拉24 h和36 h下降，差异有显著性意义(P < 0.05)。在白细胞介素1β为0.8 μg/L时，与静态对照组相比，CollagenⅠα1的表达在5%，10%，15%牵拉12 h时没有变化，而在牵拉24 h和36 h时降低，差异有显著性意义(P < 0.05)。相比于未添加白细胞介素1β时，0.2，0.4，0.8 μg/L白细胞介素1β单独作用12 h角膜成纤维细胞CollagenⅠα1的表达显著性降低(P < 0.05)。 2.2 白细胞介素1β与机械牵拉对角膜成纤维细胞CollagenⅠα2的影响白细胞介素1β与机械牵拉共同作用于角膜成纤维细胞，实时荧光定量PCR检测CollagenⅠα2 mRNA的表达，见图2。未添加白细胞介素1β时，与静态对照组相比，5%，10%，15%分别牵拉12 h使CollagenⅠα2表达降低，差异有显著性意义(P < 0.05)；牵拉24 h和36 h时，CollagenⅠα2的表达在5%牵拉时没有变化，在10%，15%牵拉时表达下降，差异有显著性意义(P < 0.05)。在白细胞介素1β为0.2，0.4 μg/L时，与静态对照组相比，CollagenⅠα2的表达在5%牵拉12 h时没有变化，而在10%，15%牵拉12 h时降低，差异有显著性意义(P < 0.05)；与静态对照组相比，CollagenⅠα2的表达在5%，10%，15%牵拉24 h和36 h下降，差异有显著性意义(P < 0.05)。在白细胞介素1β为0.8 μg/L时，与静态对照组相比，CollagenⅠα2的表达在5%，10%牵拉12 h时没有变化，而在15%牵拉12 h时降低，差异有显著性意义(P < 0.05)；与静态对照组相比，CollagenⅠα2的表达在5%，10%，15%牵拉24 h和36 h下降，差异有显著性意义(P < 0.05)。 2.3 白细胞介素1β与机械牵拉对角膜成纤维细胞Lumican的影响白细胞介素1β与机械牵拉共同作用于角膜成纤维细胞，实时荧光定量PCR检测Lumican mRNA的表达，见图3。未添加白细胞介素1β时，与静态对照组相比，5%牵拉12，24，36 h使Lumican表达升高，差异有显著性意义(P < 0.05)；10%和15%牵拉12 h使Lumican表达显著性降低(P < 0.05)，而牵拉24 h和36 h使Lumican表达升高，差异有显著性意义(P < 0.05)。在白细胞介素1β为0.2 μg/L时，5%牵拉12，24，36 h使角膜成纤维细胞Lumican的表达升高，差异有显著性意义(P < 0.05)；10%和15%牵拉在12 h使角膜成纤维细胞Lumican的表达显著性降低，而在24 h和36 h使Lumican的表达升高，差异有显著性意义(P < 0.05)。在白细胞介素1β为0.4 μg/L时，5%和10%牵拉12 h使角膜成纤维细胞Lumican的表达没有变化，15%牵拉使Lumican的表达降低，差异有显著性意义(P < 0.05)；5%，10%，15%牵拉24 h和36 h使Lumican的表达升高，差异有显著性意义(P < 0.05)。在白细胞介素1β为0.8 μg/L时，5%，10%，15%牵拉12 h使角膜成纤维细胞Lumican的表达没有变化，5%，10%，15%牵拉24 h和36 h使Lumican的表达升高，差异有显著性意义(P < 0.05)。当白细胞介素1β单独作用时，与0 μg/L时相比，Lumican在12 h和36 h的表达不随白细胞介素1β的质量浓度而变化；单独作用24 h，在白细胞介素1β质量浓度为0.2，0.4 μg/L时，Lumican的表达升高，差异有显著性意义(P < 0.05)。"

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