骨髓来源细胞表达的MMP-2/MMP-13在脉络膜新生血管发生发展中的作用

Effect of MMP-2/MMP-13 Expression by Bone Marrow-derived Cells in Development of Choroidal Neovascularization

目的探讨骨髓来源细胞(BMCs)基质金属蛋白酶(MMP)的动态表达及其与脉络膜新生血管(CNV)发生、发展的关系。方法选取成功接受骨髓移植的24只C57BL/6J绿色荧光蛋白(GFP)嵌合体小鼠为观察组,未接受骨髓移植的48只C57BL/6J小鼠为对照组。两组均予激光光凝照射以诱发CNV,利用Western blot法及明胶酶谱法检测对照组在激光诱发前、诱发后1、3、5、7、10、14、28 d时MMP-2/MMP-13的表达情况,利用免疫荧光染色检测观察组CNV中GFP阳性细胞(即移植的骨髓细胞)是否表达MMP-2/MMP-13,并进行定量分析。结果 Western blot法、明胶酶谱法、免疫荧光均显示在激光诱发早期两组CNV中MMP-2的表达即开始快速增加,在诱导第3天表达量最高、活性最强,且观察组BMCs表达MMP-2也在此时达到高峰,占总表达量的63.21%,随后MMP-2表达量开始下降;在激光诱发早期两组CNV中MMP-13的表达缓慢增加,在诱导后第7天表达量最高、活性最强,且观察组BMCs表达MMP-13亦达最高峰,占总表达量的77.87%,随后下降。结论 BMCs可能作为调控靶点改变MMP-2/MMP-13的表达,进而影响血管细胞的出芽、移行、凋亡及CNV纤维化。

Objective To investigate matrix metalloproteinase （MMP） dynamic expression by bone marrow-derived cells （BMCs） and its relationship with pathogenesy and development of choroidal neovascularization （CNV）. Methods A total of 24 C57BL/6J green fluorescent protein （GFP） of allophenic mice, who underwent bone marrow transplantation suc- cessfully, were selected as observation group, while other 48 C57BL/6J mice without hone marrow transplantation were select- ed as control group. CNV was induced by irradiating with laser photocoagulation for all mice. In control group, MMP-2/MMP- 13 expressions were detected by Western blot and gelatin zymography method before induction, and at 1st , 3rd , 5th , 7th , 10th , 14th and 28th d after induction; In observation group, immunofluorescence staining was used to identify whether or not MMP-2/ MMP-13 was expressed by GFP -positive BMCs in CNV, and quantitative analysis was also performed. Results Western blot, gelatin zymography and immunofluorescence staining methods all showed that MMP-2 expressions were rapidly increased in the early laser stage of CNV patients in two groups, and the expressions of BMCs and MMP-2 reached the highest values and strongest activity at 3rd d after induction; the highest value of MMP-2 expression in observation group occupied 63.21% of the total expression; and then MMP-2 expression began to decline. MMP-13 expression showed slowly increases at the early laser stage of CNV patients in two groups, and the highest expression and strongest activity were found at 7th d after induction; the highest values of MMP-13 expression in CNV patients in two groups and in observation group occupied 77.87% of the total ex- pression; and then MMP-13 expression began to decline. Conclusion BMCs can be used as target point to regulate MMP-2/ MMP-13 expression, and it can further affect pullulation, transmigration, apoptosis and fibration of CNV.