College of Pharmacy, Taipei Medical University, Taipei, Taiwan.
J Control Release. 2010 Oct 1;147(1):76-83. doi: 10.1016/j.jconrel.2010.06.006. Epub 2010 Jun 16.
Stromal keratocyte apoptosis triggered by epithelial injury is one mechanism of corneal disorders. A model of epithelial injury by epithelial debridement is established, and keratocyte apoptosis is evidenced by DNA fragmentation and cellular morphological changes in the anterior stroma underlying the injured epithelium. Delivery of plasmid (pCMV-bcl-x(L)-eGFP) encoding an anti-apoptotic gene, the bcl-x(L) with a nano-carrier, polymeric micelles (PM) via eye drop to cornea after epithelial debridement, the mRNA level of bcl-x(L) was significantly increased (2.2-fold, P<0.05) at 48 h and the eGFP mRNA was detected (4571.7 ± 1194.5 copies/μg total RNA). The bcl-x(L)-eGFP fusion protein was also detected in wounded cornea at 48 h after delivery, accompanying with decreased DNA fragmentation and lower caspase-3 activity (P<0.05). In conclusion, eye drop of pCMV-bcl-x(L)-eGFP/PM reduced corneal apoptosis following epithelial debridement.
上皮损伤引发基质角质细胞凋亡是角膜病变的机制之一。通过上皮清创术建立上皮损伤模型，损伤上皮下前基质中可见到角质细胞 DNA 片段化和细胞形态学改变，证实角质细胞凋亡。上皮清创术后，通过滴眼将编码抗凋亡基因 bcl-x(L)的质粒（pCMV-bcl-x(L)-eGFP）与纳米载体聚合物胶束（PM）递送至角膜，48 h 时 bcl-x(L)的 mRNA 水平显著增加（2.2 倍，P<0.05），并检测到 eGFP mRNA（4571.7±1194.5 拷贝/μg 总 RNA）。递送后 48 h 在受伤角膜中也检测到 bcl-x(L)-eGFP 融合蛋白，同时 DNA 片段化减少，半胱天冬酶-3 活性降低（P<0.05）。结论：pCMV-bcl-x(L)-eGFP/PM 滴眼可减少上皮清创术后角膜凋亡。
