Adenoviral gene transfer of BMP-7, Id2, or Id3 suppresses injury-induced epithelial-to-mesenchymal transition of lens epithelium in mice
Adenoviral gene transfer of BMP-7, Id2, or Id3 suppresses injury-induced epithelial-to-mesenchymal transition of lens epithelium in mice
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DOI：
10.1152/ajpcell.00306.2005
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发表时间：
2006-01-01
影响因子：
5.5
通讯作者：
Ooshima, A
中科院分区：
文献类型：
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作者：
Saika, S;Ikeda, K;Ooshima, A
我们研究了腺病毒介导的骨形态发生蛋白-7 （BMP-7）和分化抑制剂2和3 （Id2和Id3）的表达对小鼠晶状体上皮损伤诱导的上皮-间质转化（EMT）的影响。已知Id2和Id3被BMP-7上调，并拮抗Smad2/3信号。采用Cre-LoxP系统腺病毒基因转移。在全身麻醉和表面麻醉下，将3微升腺病毒溶液（2 × 10(7) PFU/ μ l）注射到成年雄性C57BL/6小鼠（n = 144）的右晶状体中。实验组使用cre -腺病毒（Cre-Ad）和编码mBMP-7、mId2或mId3的载体的混合物。对照晶状体单独用Cre-Ad处理。在5天或10天的愈合间隔后，将动物杀死，然后我们进行组织学处理或从晶状体中提取RNA。RT-PCR、real-time RT-PCR和免疫组织化学显示每个引入基因在晶状体中的表达。外源BMP-7上调损伤晶状体中Id2和Id3的表达，Id2或Id3的基因导入也上调BMP-7的表达。通过组织学和α -平滑肌肌动蛋白和胶原的表达模式评估，BMP-7、Id2或Id3的基因转移延迟了晶状体上皮细胞损伤诱导的EMT，这与Smad2 COOH-末端磷酸化的减少有关。BMP-7、Id2或Id3的基因转移延迟了损伤诱导的小鼠晶状体上皮细胞的EMT和随后被纤维组织封闭的荚膜破裂。
We have examined the effect of adenovirus-mediated expression of bone morphogenic protein-7 (BMP-7) and inhibitors of differentiation 2 and 3 (Id2 and Id3) on injury-induced epithelial-to-mesenchymal transition (EMT) of lens epithelium in mice. Id2 and Id3 are known to be upregulated by BMP-7 and to antagonize Smad2/3 signaling. The Cre-LoxP system adenoviral gene transfer was used. Three microliters of adenoviral solution (2 X 10(7) PFU/mu l) were injected into the right lens of adult male C57BL/6 mice (n = 144) at the time of capsular injury induced using a hypodermic needle under both general and topical anesthesia. A mixture of Cre-adenovirus (Cre-Ad) and vector encoding mBMP-7, mId2, or mId3 was administered in a test group. Control lenses were treated with Cre-Ad alone. After healing intervals of 5 or 10 days, the animals were killed and then we performed histological processes or RNA extraction from the lens. RT-PCR, real-time RT-PCR, and immunohistochemistry showed expression of each introduced gene in the lens. Exogenous BMP-7 upregulated expression of Id2 and Id3 in injured lenses, and gene introduction of Id2 or Id3 also upregulated BMP-7 expression. Gene transfer of BMP-7, Id2, or Id3 delayed injury-induced EMT of the lens epithelial cells as evaluated by histology and expression patterns of alpha-smooth muscle actin and collagens in association with reduction of Smad2 COOH- terminal phosphorylation. Gene transfer of BMP-7, Id2, or Id3 delayed injury-induced EMT of lens epithelial cells and subsequent sealing of the capsular break with fibrous tissue in mice.