Cationic star-shaped polymer as an siRNA carrier for reducing MMP-9 expression in skin fibroblast cells and promoting wound healing in diabetic rats.
Cationic star-shaped polymer as an siRNA carrier for reducing MMP-9 expression in skin fibroblast cells and promoting wound healing in diabetic rats.
复制标题
阳离子星形聚合物作为siRNA载体降低皮肤成纤维细胞中MMP-9的表达并促进糖尿病大鼠伤口愈合
DOI：
10.2147/ijn.s66368
复制
发表时间：
2014
影响因子：
8
通讯作者：
Zhang LM
中科院分区：
文献类型：
--
作者：
Li N;Luo HC;Yang C;Deng JJ;Ren M;Xie XY;Lin DZ;Yan L;Zhang LM
基质金属蛋白酶-9 （MMP-9）的过度表达对糖尿病患者的皮肤创面愈合过程有害。本研究旨在探讨由β-环糊精（β-CD）核心和聚氨基胺树突臂（β-CD-[D3]7）组成的阳离子星形聚合物能否作为小干扰RNA （siRNA）的基因载体，降低MMP-9的表达，促进糖尿病创面愈合。采用3-（4,5-二甲基噻唑-2-基）-2,5-二苯基溴化四唑试验（MMT）法研究了β-CD-(D3)7对大鼠CRL1213皮肤成纤维细胞系的细胞毒性。用共聚焦显微镜和流式细胞术检测β-CD-(D3)7/ mmp -9小干扰RNA （siRNA）复合物的转染效率。采用定量实时（RT）聚合酶链反应测定β-CD-(D3)7/MMP-9- sirna复合物转染后MMP-9的基因表达情况。将β-CD-(D3)7/MMP-9-siRNA复合物注射于链脲佐菌素诱导的糖尿病大鼠创面。伤后第4天和第7天测量伤口闭合。β-CD-(D3)7在成纤维细胞中表现出较低的细胞毒性，且容易与MMP-9-siRNA形成复合物。β-CD-(D3)7/MMP-9- sirna复合物容易被成纤维细胞占用，导致MMP-9基因表达下调（P<0.01）。动物实验结果显示，β-CD-(D3)7/MMP-9-siRNA复合物对糖尿病大鼠损伤后第7天伤口愈合有促进作用（P<0.05）。β-CD-(D3)7可作为MMP-9- sirna的有效载体，降低糖尿病大鼠皮肤成纤维细胞中MMP-9的表达，促进创面愈合。
Excessive expression of matrix metalloproteinase-9 (MMP-9) is deleterious to the cutaneous wound-healing process in the context of diabetes. The aim of the present study was to explore whether a cationic star-shaped polymer consisting of β-cyclodextrin (β-CD) core and poly(amidoamine) dendron arms (β-CD-[D3]7) could be used as the gene carrier of small interfering RNA (siRNA) to reduce MMP-9 expression for enhanced diabetic wound healing. The cytotoxicity of β-CD-(D3)7 was investigated by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay (MMT) method in the rat CRL1213 skin fibroblast cell line. The transfection efficiency of β-CD-(D3)7/MMP-9-small interfering RNA (siRNA) complexes was determined by confocal microscopy and flow cytometry. Quantitative real time (RT) polymerase chain reaction was performed to measure the gene expression of MMP-9 after the transfection by β-CD-(D3)7/MMP-9-siRNA complexes. The β-CD-(D3)7/MMP-9-siRNA complexes were injected on the wounds of streptozocin-induced diabetic rats. Wound closure was measured on days 4 and 7 post-wounding. β-CD-(D3)7 exhibited low cytotoxicity in fibroblast cells, and easily formed the complexes with MMP-9-siRNA. The β-CD-(D3)7/MMP-9-siRNA complexes were readily taken up by fibroblast cells, resulting in the downregulation of MMP-9 gene expression (P<0.01). Animal experiments revealed that the treatment by β-CD-(D3)7/MMP-9-siRNA complexes enhanced wound closure in diabetic rats on day 7 post-wounding (P<0.05). β-CD-(D3)7 may be used as an efficient carrier for the delivery of MMP-9-siRNA to reduce MMP-9 expression in skin fibroblast cells and promote wound healing in diabetic rats.