携带ad-vegf165转基因细胞丝素膜诱导角膜血管化和ad-hil-17f抑制作用-silk fibroin membrane carrying ad - vegf 165 transgenic cells induces corneal vascularization and ad - hil - 17f inhibition.docxVIP

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2018-08-07 发布于上海
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携带ad-vegf165转基因细胞丝素膜诱导角膜血管化和ad-hil-17f抑制作用-silk fibroin membrane carrying ad - vegf 165 transgenic cells induces corneal vascularization and ad - hil - 17f inhibition.docx

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携带ad-vegf165转基因细胞丝素膜诱导角膜血管化和ad-hil-17f抑制作用-silk fibroin membrane carrying ad - vegf 165 transgenic cells induces corneal vascularization and ad - hil - 17f inhibition

IV IV 携带 Ad-VEGF165 转基因细胞再生丝素膜诱导角膜血管化和 Ad-hIL-17F 抑制作用英文摘要 indirect immunofluorescence methods. The growth-suppressing effect of Ad-hIL-17F on the ECV304 cells was assessed by MTT. The transcription and expression of VEGF in ECV304 cells effect of Ad-hIL-17F was determined by ELISA, real-time quantitative PCR. Regenerated silk fibroin films modified by transgenes cells were implanted in corneal stroma. The in vivo enhanced growth-suppressing effect of Ad-hIL-17F on corneal neovessels by subconjunctival injection at one week after operation. The conjunctival congestion and the area of neovessels were recorded. Expression of CD34 and VEGF in corneal stroma were detected by immunohistochemistry. RESULTS: (1) corneal epithelium cells grow well in regenerated silk protein thin film. Rabbit corneal stroma implanted with regenerated silk protein thin film remain transparent with no obvious neovascularization nor implant falling off, no infection and no inflammatory reactions in the anterior chamber of the eye. (2)In the culture plate of Ad-VEGF165 transgenic corneal epithelium cells, the expression of VEGF increases, meanwhile, the expression of neovascularization related growth factors such as Ang1、EGF、TGF-β etc. also increased. After the implantation of regenerated silk protein film loaded with Ad-VEGF165 transgenic corneal epithelium cells, cornea healing reaction is strong, inflammatory cells infiltrates, collagen hyperplasia, the expression of neovascularization related cell factors such as VEGF、MMP-2、TGF-β increase. The implantion of regenerated silk protein film loaded with Ad-VEGF165 transgenic corneal epithelium cells can induce neovascularization in cornea and the new vessels can sustain more than one month. (3) Sequencing results indicate that the sequence of hIL-17F is correct. The expression of IL-17F in QBI-293A cells are detected by RT-PCR and indirect immuno-fluorescence methods. Ad-hIL-17F suppresses the growth of ECV304 cells. It suppre