NF-E2相关因子2对糖尿病模型小鼠视网膜内细胞及血管的保护作用

发布时间：2018-06-02 16:24

本文选题：糖尿病 + 视网膜病变　；参考：《解剖学报》2017年05期

【摘要】：目的探讨NF-E2相关因子2(Nrf2)对糖尿病视网膜细胞及血管的保护作用。方法 8周龄雄性Nrf2~(+/+)(野生型)和Nrf2~(-/-)C57BL/6小鼠60只随机分为模型组和对照组,共4组,每组各15只。模型组小鼠腹腔注射链脲佐菌素(STZ,45mg/kg),连续注射5 d;对照组小鼠注射同体积的枸橼酸缓冲液。模型组和对照组小鼠初次注射后每周测空腹血糖及体重,至第10周取视网膜。采用免疫印迹法检测视网膜内Nrf2激活的下游蛋白血红素加氧酶-1(HO-1)的表达情况;免疫荧光染色检测具有多种剪接形式的RNA结合蛋白(RBPMS)阳性的视网膜节细胞(RGCs)及胆碱乙酰转移酶(Ch AT)阳性的无长突细胞(ACs)并计数;采用过碘酸-希夫和苏木素染色,观察视网膜内无细胞毛细血管并进行定量分析。结果 STZ诱导1周后,野生型和Nrf2~(-/-)模型组小鼠血糖急剧升高,体重开始显著下降,随着周龄的增长体重无明显增加;对照组小鼠血糖值无明显升高,体重呈正常缓慢增长趋势。STZ诱导糖尿病10周后,RBPMS和Ch AT染色发现,Nrf2~(~(-/-))糖尿病小鼠视网膜内RGCs和ACs数量显著降低(P0.05);过碘酸-希夫和苏木素染色显示,Nrf2~(-/-)糖尿病小鼠视网膜内出现无细胞毛细血管;免疫印迹法显示,野生型小鼠模型组中Nrf2激活的下游蛋白HO-1表达升高。结论 Nrf2对糖尿病视网膜细胞及血管具有保护作用,其可能是通过诱导HO-1上调发挥作用。
[Abstract]:Objective to investigate the protective effect of NF-E2 related factor 2 nrf 2 on diabetic retinal cells and blood vessels. Methods Sixty 8-week-old male Nrf2G (wild type) and Nrf2~(-/-)C57BL/6 mice were randomly divided into 4 groups (15 in each group) and 4 in model group and control group. The mice in the model group were injected with STZ 45 mg / kg for 5 days and the control group was injected with the same volume of citrate buffer. Fasting blood glucose and body weight were measured weekly after the first injection in the model group and control group, and retina was removed at the 10th week. The expression of heme oxygenase-1 (HO-1), a downstream protein activated by Nrf2 in the retina, was detected by Western blot. Immunofluorescence staining was used to detect and count the RGCs of retinal ganglion cells (RGCs) and choline acetyltransferase (Chat) positive cells of retinal ganglion cells (RGCs) with multiple splicing forms of RNA binding protein (RNA binding protein), and to count them with periodate Schiff and hematoxylin staining. The capillary vessels in the retina were observed and quantitatively analyzed. Results after 1 week of STZ induction, the blood glucose of the wild type and Nrf2 + / -r / -group increased sharply, the body weight began to decrease significantly, and the body weight did not increase with the increase of week age, but the blood sugar value of the control group did not increase significantly. STZ-induced diabetes mellitus 10 weeks later, the number of RGCs and ACs in retina of diabetic mice decreased significantly (P 0.05), and periodate Schiff and hematoxylin staining showed that the visual reticulum of diabetic mice was nrf2r-r-1. Acellular capillaries appeared in the membrane; Western blotting showed that the expression of downstream protein HO-1 activated by Nrf2 was increased in the wild-type mouse model group. Conclusion Nrf2 has protective effect on diabetic retinal cells and blood vessels, which may play a role by inducing up-regulation of HO-1.
【作者单位】：北京大学医学部人体解剖学与组织学胚胎学系;
【基金】：国家自然科学基金青年基金(81200691)
【分类号】：R587.2;R774.1

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