MMP-2在糖尿病模型大鼠视网膜组织中的表达

doi:10.13481/j.1671-587x.20170512

摘要/Abstract

摘要： 目的：采用腹腔注射链脲佐菌素（STZ）法复制糖尿病大鼠模型，观察糖尿病模型大鼠不同时期视网膜组织中基质金属蛋白酶2（MMP-2）表达水平，阐明MMP-2在糖尿病大鼠糖尿病视网膜病变（DR）中的作用。方法：雌性SD大鼠分为正常对照组（n=24）、4周模型组（n=30）、6周模型组（n=30）和8周模型组（n=30），模型组大鼠连续5 d腹腔注射小剂量STZ复制糖尿病大鼠模型，正常对照组大鼠注射等量的柠檬酸钠溶液。4、6和8周时检测各组大鼠体质量和血糖水平。末次检测后麻醉获取大鼠右眼视网膜组织，采用RT-PCR和Western blotting法检测MMP-2 mRNA及蛋白表达水平，免疫组织化学染色观察大鼠视网膜组织中MMP-2表达情况。结果：造模前，各组大鼠体质量和禁食12 h的空腹血糖值比较差异均无统计学意义（P > 0.05）。4周模型组大鼠死亡3只，6周时死亡5只，8周时死亡7只，存活动物继续进行实验。各模型组大鼠体质量均明显低于同时期正常对照组（P < 0.05或P < 0.01），而空腹血糖值明显升高（P < 0.05或P < 0.01）。各模型组大鼠视网膜组织中MMP-2 mRNA和蛋白表达水平均明显高于正常对照组（P < 0.05）。正常对照组大鼠视网膜结构层次清晰分明、细胞排列整齐，可见单层排列的神经节细胞，核圆形或椭圆形状，核较大；与正常对照组比较，模型组大鼠视网膜组织结构松散，内核层及视杆细胞层细胞外界膜模糊，数量减少。MMP-2阳性细胞可见棕黄色颗粒沉淀，在神经节细胞层胞浆和血管内皮细胞尤为集中。4、6及8周模型组MMP-2阳性表达水平均高于正常对照组（P < 0.05）。结论：MMP-2在糖尿病模型大鼠视网膜上表达，并随着糖尿病大鼠模型维持时间的延长呈递增趋势，说明MMP-2与DR有关。

关键词: 糖尿病视网膜病变, 基质金属蛋白酶2, 链脲佐菌素, 糖尿病大鼠模型

Abstract: Objective: To establish the rat diabetic models by introperitoneal injection of streptozotocin(STZ) to observe the expressions of MMP-2 in retina tissue of the diabetic rats at different periods,and to clarify the effect of MMP-2 in the diabetic retiropathy(DR) of the diabetic rats. Methods: The femal SD rats were divded into normal control group(n=24), 4-week model group(n=30), 6-week model group(n=30) and 8-week model group(n=30).The rats in model groups were intraperitoneally injected with STZ for consecutive 5 d.The rats in normal control group were injected with sodium citrate solution at the same volume. The body weights and blood glucose levels of the rats in each group were measured at 4,6, and 8 weeks.The retina of each rat was removed; RT-PCR and Western blotting methods were used to detect the expression levels of MMP-2 mRNA and protein,and the immunohistochemistry was conducted to observe the morphology. Results: The body weights and blood glucose levels of the rats in each group had no differences(P>0.05) before modeling.Three rats died in 4-week model group, 5 rats died at 6 weeks and 7 rats died at 8 weeks.The body weights of the rats in model group were significantly lower than those in normal control group at the same time(P<0.05 or P<0.01),and the fasting blood glucose levels were signifieantly increased(P<0.05 or P<0.01).The expression levels of MMP-2 mRNA and protein in the retina tissue of the rats in model group were significantly higher than those in normal control group(P<0.05). In normal control group, the retinal structure was clear and the cells were arranged in order, and the ganglion cells were arranged in a single layer; in model group, the retinal tissue structure was loose, the number of ganglion cells were significantly reduced,the inner nuclear layer and rod cell layer cell membrane outside were fuzzy. The MMP-2 positive cells were found to be brown yellow granules, especially in the cytoplasm of ganglion cells and vascular endothelial cells. The positive expression levels of MMP-2 in model group were higher than those in normal control group at 4,6,and 8 weeks. Conclusion: MMP-2 can express in the retina tissue of the diabetic rats.

Key words: matrix metalloproteinase-2, streptozotocin, diabetic retinopathy

中图分类号:

R774.1

杨洁, 马英慧, 崔秀成, 王红梅, 董微丽, 谭小波. MMP-2在糖尿病模型大鼠视网膜组织中的表达[J]. 吉林大学学报(医学版), 2017, 43(05): 923-927.

YANG Jie, MA Yinghui, CUI Xiucheng, WANG Hongmei, DONG Weili, TAN Xiaobo. Expression of MMP-2 in retina tissue of diabetic retinopathy model rats[J]. Journal of Jilin University Medicine Edition, 2017, 43(05): 923-927.

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