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塞来昔布对三维角膜胶原降解抑制作用的分子机制

Molecular mechanism of the inhibition effect of Celecoxib on corneal collagen degradation in three dimensions.

作者信息

Zhang Wen-Song, Hao Ji-Long, Wang Shuang, Bi Miao-Miao, Zhang Hong, Zhou Hong-Yan

机构信息

Department of Ophthalmology, the Second Hospital of Jilin University, Changchun 130041, Jilin Province, China.

出版信息

Int J Ophthalmol. 2012;5(4):434-9. doi: 10.3980/j.issn.2222-3959.2012.04.06. Epub 2012 Aug 18.

Abstract

AIM

To clarify the molecular mechanism of Celecoxib on corneal collagen degradation and corneal ulcer.

METHODS

Rabbit corneal fibroblasts were harvested and suspended in serum-free MEM. Type I collagen, DMEM, collagen reconstitution buffer and corneal fibroblast suspension were mixed on ice. The resultant mixture solidify in an incubator, after which test reagents and plasminogen was overlaid and the cultures were returned to the incubator. The supernatants from collagen gel incubations were collected and the amount of hydroxyproline in the hydrolysate was measured. Immunoblot analysis of MMP1, 3 and TIMP1, 2 was performed. MMP2, 9 was detected by the method of Gelatin zymography. Cytotoxicity Assay was measured.

RESULTS

Celecoxib inhibited corneal collagen degradation in a dose and time manner; Celecoxib inhibited the IL-1ß induced increases in proMMP1, 2, 3, 9 and active MMP1, 2, 3, 9 in a concentration-depended manner. Celecoxib can also inhibit the IL-1ß induced increases in the TIMP1, 2.

CONCLUSION

Celecoxib can inhibit corneal collagen degradation induced by IL-1β, this effect is the consequence of the reduction of MMP1, 2, 3, 9 and TIMP1, 2. The results of the present study provide new insight into Celecoxib in corneal ulcer treatment.

摘要

目的

阐明塞来昔布对角膜胶原降解及角膜溃疡的分子机制。

方法

收获兔角膜成纤维细胞并悬浮于无血清的MEM中。将I型胶原、DMEM、胶原重构缓冲液和角膜成纤维细胞悬液在冰上混合。所得混合物在培养箱中凝固,之后覆盖测试试剂和纤溶酶原,再将培养物放回培养箱。收集胶原凝胶孵育的上清液并测量水解产物中羟脯氨酸的量。进行MMP1、3和TIMP1、2的免疫印迹分析。通过明胶酶谱法检测MMP2、9。进行细胞毒性测定。

结果

塞来昔布以剂量和时间依赖方式抑制角膜胶原降解;塞来昔布以浓度依赖方式抑制IL-1β诱导的proMMP1、2、3、9和活性MMP1、2、3、9增加。塞来昔布还可抑制IL-1β诱导的TIMP1、2增加。

结论

塞来昔布可抑制IL-1β诱导的角膜胶原降解,这种作用是MMP1、2、3、9和TIMP1、2减少的结果。本研究结果为塞来昔布治疗角膜溃疡提供了新的见解。

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