PURPOSE: The effects of small interfering (si)RNA of nuclear factor kappa B (NF-kappaB) on the development of posterior capsule opacification (PCO) were investigated both in vitro and in vivo in rabbits. METHODS: After application of p105 NF-kappaB siRNA to lens epithelial cells (LECs), Western blot analyses were performed to detect p105 and p50 NF-kappaB and a scratch assay was used to determine cell migration. In the capsular bag model, immunocytochemistry was performed to determine expression of p50 NF-kappaB and Western blot analyses for the presence of epithelial-to-mesenchymal transition (EMT) markers. Two sequences of p105 NF-kappaB siRNA were used in cataract surgery in 15 New Zealand White rabbits. PCO grading was conducted by slit lamp biomicroscopy and a computer-based PCO grading program. One month after surgery, the eyes of the rabbits were enucleated, and sections were prepared for examination of the posterior capsule and other ocular tissues by light microscopy. RESULTS: Application of p105 NF-kappaB siRNA to LECs decreased p105 NF-kappaB and p50 NF-kappaB expression, and migration of LECs was shown to be inhibited on the scratch assay. In the capsular bag model, the LEC count was significantly decreased, and immunocytochemistry showed reduced p50 NF-kappaB expression on the posterior capsule. EMT markers were significantly decreased after application of p105 NF-kappaB siRNA in the capsular bag model. In the in vivo study in rabbits, p105 NF-kappaB siRNA effectively decreased PCO, as determined by both slit lamp examination and the PCO grading program. CONCLUSIONS: NF-kappaB seems to be related to migration and proliferation of LECs. NF-kappaB siRNA was effective in inhibiting the migration and proliferation of LECs in vitro and decreased PCO formation after cataract surgery in an in vivo rabbit model.
PURPOSE: The effects of small interfering (si)RNA of nuclear factor kappa B (NF-kappaB) on the development of posterior capsule opacification (PCO) were investigated both in vitro and in vivo in rabbits. METHODS: After application of p105 NF-kappaB siRNA to lens epithelial cells (LECs), Western blot analyses were performed to detect p105 and p50 NF-kappaB and a scratch assay was used to determine cell migration. In the capsular bag model, immunocytochemistry was performed to determine expression of p50 NF-kappaB and Western blot analyses for the presence of epithelial-to-mesenchymal transition (EMT) markers. Two sequences of p105 NF-kappaB siRNA were used in cataract surgery in 15 New Zealand White rabbits. PCO grading was conducted by slit lamp biomicroscopy and a computer-based PCO grading program. One month after surgery, the eyes of the rabbits were enucleated, and sections were prepared for examination of the posterior capsule and other ocular tissues by light microscopy. RESULTS: Application of p105 NF-kappaB siRNA to LECs decreased p105 NF-kappaB and p50 NF-kappaB expression, and migration of LECs was shown to be inhibited on the scratch assay. In the capsular bag model, the LEC count was significantly decreased, and immunocytochemistry showed reduced p50 NF-kappaB expression on the posterior capsule. EMT markers were significantly decreased after application of p105 NF-kappaB siRNA in the capsular bag model. In the in vivo study in rabbits, p105 NF-kappaB siRNA effectively decreased PCO, as determined by both slit lamp examination and the PCO grading program. CONCLUSIONS: NF-kappaB seems to be related to migration and proliferation of LECs. NF-kappaB siRNA was effective in inhibiting the migration and proliferation of LECs in vitro and decreased PCO formation after cataract surgery in an in vivo rabbit model.
Authors: Andrea Hoffmann; Yusen Huang; Rinako Suetsugu-Maki; Carol S Ringelberg; Craig R Tomlinson; Katia Del Rio-Tsonis; Panagiotis A Tsonis Journal: Mol Med Date: 2012-05-09 Impact factor: 6.354