高脂联合高糖饲养小鼠糖尿病性角膜病变模型的建立

摘要

目的：研究利用高脂联合高糖饲养C57 BL/6小鼠建立糖尿病小鼠角膜病变模型。　　方法：建立利用高脂联合高糖饲养C57 BL/6小鼠的糖尿病模型,在成模2~12 mo分别行1%虎红染色检查角膜上皮完整性；角膜上皮刮除后,荧光素钠染色观察比较角膜上皮的愈合速度；病理检测角膜的组织形态和结构的变化。　　结果：高脂联合高糖饲养C57 BL/6小鼠2 mo左右逐渐表现出糖尿病多饮、多食、多尿、体质量下降等典型症状,其血糖稳定维持在较高水平(≥18mmol/L),体质量较正常小鼠明显偏低。虎红染色发现,成模2 mo的小鼠角膜即已出现点状着色,且随着病程的延长,染色面积和程度逐渐加重,到成模12 mo几乎呈全角膜着色。在上皮愈合速度方面,刮除角膜上皮后,成模2 mo的小鼠角膜上皮在40 h即完全愈合(与正常小鼠相似),成模3 mo小鼠上皮愈合时间为120h,成模4,6,12mo上皮完全愈合的时间为144 h左右,其中成模12 mo的小鼠在96~120 h上皮缺损的范围再次扩大,后逐渐缩小愈合,呈现反复现象。　　结论：高脂联合高糖饲养诱导的糖尿病小鼠角膜,表现出角膜上皮损害,角膜上皮损伤后延迟愈合等症状,说明其可以作为研究糖尿病角膜病变的动物模型。%AIM: To discuss the establishment of immediate diabetic keratopathy animal model of C57BL/6 mouse induced by ahigh-fat and high-glucose diet. METHODS: Diabetes mellitus was induced by a high-fat and high-glucose diet in C57BL/6 mouse. 1% rose bengal was stained on the cornea to examine the integrality of the corneal epithelium at 2 ~ 12mo after completion of the model. Corneal epithelial wound healing was observed using a vivo epithelial debridement model which was dyed by sodium fluorescein. Corneal morphology histology was examined by pathological methods. RESULTS: The high-fat and high-glucose diet C57BL/6 mouse in 2mo had showed general symptoms of diabetes: polydipsia, polyphagia, polyuria, weight loss etc. The model had a steady-state high glucose (≥18mmol/L), also the weight was lower compared with normal control mouse. 1% rose bengal corneal staining had dot coloring at 2mo after completion of the model, the stained area and extent were gradually increased with the extension of the duration of diabetes, almost all the cornea was stained at 12mo after completion of the model. With the passage of time into a mold, the cornea epithelial healing time become longer: 2mo was about 40h;3mo was about 120h; 4, 6, 12mo was about 144h;the coloboma were gradually increased at 12mo after completion of the model, then the area was reduced gradually until complete healing, the time was 96~120h, showed repeating phenomenon. CONCLUSION: The mouse were induced by high-fat and high-glucose diet can be used as animal models of diabetic keratopathy: the damage of epithelium for corneal and delay healing on epithelium and other symptoms.