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目的通过玻璃体腔注射shRNA-ANG慢病毒载体,干扰血管生成素(ANG)的表达,电镜观察糖尿病大鼠视网膜形态及分子水平检测视网膜ANG的表达,探讨ANG在糖尿病视网膜微血管病变中的作用。方法将40只大鼠随机分为空白对照组、模型对照组、空载体组及shRNA-ANG干扰组,利用链脲佐菌素(STZ)建立1型糖尿病大鼠模型,造模成功2周后,空载体组及干扰组玻璃体腔分别注射空载体及shRNA-ANG慢病毒载体,其余注射等量生理盐水。4周及14周后摘除眼球,分离视网膜,通过实时荧光定量PCR检测ANG的表达情况,电镜观察糖尿病视网膜血管病变情况。结果4周时,ANG在shRNA-ANG干扰组的表达量下降,差异具有统计学意义(P<0.05),14周时,通过电镜观察糖尿病大鼠视网膜血管超微结构发现,shRNA-ANG干扰组大鼠毛细血管超微结构受损情况较模型对照组及空载体组明显改善。结论慢病毒介导shRNA干扰糖尿病大鼠视网膜病变中ANG的表达能减轻糖尿病大鼠视网膜微血管病变程度。
Objective To observe the expression of angiopoietin (ANG) through intravitreal injection of shRNA-ANG lentiviral vector and observe the morphological and molecular level of retina by electron microscope to detect the expression of ANG in retina and explore the role of ANG in diabetic retinal microangiopathy. Methods Forty rats were randomly divided into blank control group, model control group, empty vector group and shRNA-ANG interference group. Type 1 diabetic rat model was established by streptozotocin (STZ) , Empty vector group and interference group vitreous cavity were injected empty vector and shRNA-ANG lentiviral vector, the rest injected the same amount of saline. After 4 weeks and 14 weeks, the eyeballs were removed, the retina was separated, the expression of ANG was detected by real-time fluorescence quantitative PCR, and the diabetic retinopathy was observed by electron microscope. Results At 4 weeks, the expression of ANG in shRNA-ANG knockdown group decreased significantly (P <0.05). At 14 weeks, the ultrastructure of retinal blood vessels in diabetic rats was observed by electron microscopy. The shRNA-ANG knockdown group The damage of capillary ultrastructure in rats was significantly improved compared with model control group and empty vector group. Conclusion The lentivirus-mediated shRNA interferes with the expression of ANG in retinopathy of diabetic rats and can reduce the degree of retinal microangiopathy in diabetic rats.