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目的观察Ad-siICOS-EGFP阻断ICOS共刺激通路延长大鼠角膜植片存活时间,及抑制大鼠角膜移植急性免疫排斥的作用。方法建立大鼠同种异体穿透性角膜移植模型,将其随机分为A、B、C 3组:A组为单纯角膜移植对照组;B组为Ad-siICOS-EGFP注射组;C组为Ad-EGFP注射组。术后观察排斥反应指数RI的变化并记录植片的存活时间。各移植组分别于术后5 d随机取10只大鼠,麻醉处死后,角膜冰冻切片行共聚焦显微镜观察。各组其余大鼠进行角膜植片存活时间统计。结果①各组角膜植片平均存活时间:B组角膜植片平均存活时间最长;②共聚焦显微镜观察GFP表达情况:B组及C组角膜植片可见绿色荧光表达,表达位于角膜上皮层及整个角膜基质层,A组角膜植片未见绿色荧光表达。结论移植术后即刻受体鼠球结膜下注射Ad-siICOS-EGFP,可延长大鼠角膜植片存活时间,抑制大鼠角膜移植急性免疫排斥反应。
Objective To observe the effect of Ad-siICOS-EGFP blocking ICOS costimulatory pathway to prolong the survival time of rat corneal graft and to inhibit the acute rejection of rat corneal transplantation. Methods Allogeneic penetrating keratoplasty rat models were established and randomly divided into A, B and C groups: A group was simple corneal transplantation control group; B group was Ad-siICOS-EGFP injection group; C group was Ad-EGFP injection group. The change of rejection index RI was observed after operation and the survival time of the graft was recorded. In each transplantation group, 10 rats were randomly selected on the 5th day after operation. After the anesthesia was sacrificed, the cornea frozen sections were observed by confocal microscopy. The remaining rats in each group were collected for survival time of corneal graft. Results ① The average survival time of corneal graft in each group: the longest survival time of corneal graft in group B was the longest; ② The expression of GFP was observed by confocal microscope: the green fluorescence was expressed in corneal graft in group B and group C, The whole corneal stroma, A group of corneal graft no green fluorescent expression. Conclusion Ad-siICOS-EGFP injected into rat subconjunctival immediately after transplantation can prolong the survival time of rat corneal graft and inhibit the acute rejection after corneal transplantation.