叔丁基对苯二酚激活Nrf2信号通路增强对2型糖尿病大鼠视网膜的保护作用

发布时间：2018-03-13 20:06

本文选题：叔丁基对苯二酚　切入点：糖尿病　出处：《眼科新进展》2017年03期 　论文类型：期刊论文

【摘要】：目的研究叔丁基对苯二酚(tertiary butyl hydroquinone,tBHQ)对2型糖尿病大鼠视网膜的保护作用及相关机制。方法取雄性Sprague Dawley大鼠60只,分为正常对照组(NC组)、糖尿病组(DM组)和tBHQ组。剔除饲养过程死亡大鼠,最终NC组、DM组、tBHQ组分别为12只、20只、20只大鼠纳入研究。造模采用高脂高糖饲料喂养4周后腹腔内注射链脲佐菌素诱导2型糖尿病模型,tBHQ组于造模后1周在高脂高糖饲料中添加质量分数1%tBHQ进行干预,于造模后4周和12周心脏采血,检测血清空腹血糖(fasting plasma glucose,FPG)、血清空腹胰岛素(fasting serum insulin,FINs)含量。采用免疫组织化学法及qRT-PCR法定量检测各组大鼠视网膜中Nrf2、HO-1、Bcl-2、VEGF蛋白及mRNA的分布和表达。结果3组大鼠在4周和12周的FPG水平总体比较差异有统计学意义(F分组=78.531,P=0.000);DM组及tBHQ组高于NC组,DM组12周高于4周,tBHQ组12周明显低于4周,差异均有统计学意义(均为P0.05)。3组大鼠在4周和12周的FINs水平总体比较差异有统计学意义(F分组=22.480,P=0.000);其中DM组及tBHQ组高于NC组,tBHQ组12周高于4周,差异均有统计学意义(均为P0.05)。光学显微镜显示12周时NC组无明显改变;DM组大鼠视网膜组织结构排列疏松,内、外核层排列紊乱,细胞水肿明显;tBHQ组大鼠视网膜组织结构较清晰,部分细胞水肿。4周和12周时各组中均可见Nrf2、HO-1、Bcl-2、VEGF的阳性表达。Nrf2主要分布于视网膜节细胞层、内核层,HO-1、Bcl-2主要分布于视网膜节细胞层、内核层、外核层;VEGF主要分布于神经纤维层、节细胞层和内核层。免疫组织化学及qRT-PCR检测显示,各组大鼠在造模后不同时间点视网膜中Nrf2、HO-1、Bcl-2、VEGF蛋白及mRNA的相对表达量的总体比较差异均有统计学意义(均为P0.05)。4周时,DM组Nrf2、HO-1、VEGF的表达较NC组增加,差异均有统计学意义(均为P0.05);tBHQ组Nrf2、HO-1、Bcl-2的表达较DM组增加,VEGF的表达较DM组降低,差异均有统计学意义(均为P0.05)。12周时,DM组Nrf2、HO-1、Bcl-2、VEGF的表达较NC组增加,差异均有统计学意义(均为P0.05);tBHQ组Nrf2、HO-1、Bcl-2的表达较DM组增加,VEGF的表达较DM组降低,差异均有统计学意义(均为P0.05)。12周与4周比较,DM组VEGF的表达较4周增加,HO-1较4周降低,差异均有统计学意义(均为P0.05);tBHQ组Nrf2、HO-1、Bcl-2的表达较4周增加,差异均有统计学意义(均为P0.05)。结论tBHQ对DM大鼠胰岛功能具有一定的保护作用;可促进DM大鼠视网膜组织中Nrf2、HO-1、Bcl-2的表达,降低VEGF的表达,对视网膜组织有抗氧化应激损伤、减少细胞凋亡、抑制视网膜血管增殖等作用。tBHQ可能通过Nrf2/HO-1/VEGF及Nrf2/Bcl-2途径对DM大鼠视网膜起保护作用。
[Abstract]:Objective to study the protective effect of tertiary Ding Ji butyl hydroquinone tBHQ on retina of type 2 diabetic rats. Methods 60 male Sprague Dawley rats were selected. The rats were divided into normal control group (NC group), diabetic group (DM group) and tBHQ group. Finally, the NC group and DM group were divided into 12 / 20 / 20 rats respectively. The model was fed with high fat and high sugar diet for 4 weeks, and then intraperitoneal injection of streptozotocin (STZ) was used to induce type 2 diabetes mellitus. The tBHQ group was hyperlipidemic and hyperlipidemic 1 week after the establishment of the model, the model was induced by intraperitoneal injection of streptozotocin (streptozotocin). 1 tBHQ was added to the sugar feed to intervene, Heart blood was collected at 4 and 12 weeks after modeling. The contents of serum fasting plasma and fasting serum insulin fins were measured. The distribution and expression of Nrf2HO-1 Bcl-2VEGFin and mRNA in the retina of rats in each group were detected by immunohistochemical method and qRT-PCR method. There was significant difference in FPG level between 12 weeks and 12 weeks. F group (78.531) and tBHQ group (P < 0.05) were significantly lower than those in NC group at 12 weeks and 4 weeks, respectively. There were significant differences in the FINs level between the four and twelve weeks of P0.053.The FINs levels in DM and tBHQ groups were higher than those in NC group at 12 weeks and 4 weeks, respectively, and there were significant differences between the two groups at 12 weeks and 12 weeks, respectively, and there were significant differences between the two groups at 12 weeks and 12 weeks, respectively, and there was no significant difference between DM group and tBHQ group in 12 weeks and 4 weeks after treatment. The difference was statistically significant (P 0.05). Optical microscope showed that the retinal tissue structure in NC group was loose, the inner and outer nuclear layers were disordered, and the retinal tissue structure was clear in TBHQ group at 12 weeks. The positive expression of Nrf2HO-1Bcl-2VEGF2 was found in the retinal ganglion cell layer at the 4th and 12th week of edema. The HO-1 Bcl-2 was mainly distributed in the retinal ganglion cell layer, the nuclear layer and the outer nuclear layer were mainly distributed in the nerve fiber layer. Ganglion cell layer and nuclear layer. Immunohistochemical and qRT-PCR detection. There were significant differences in the expression of Nrf2HO-1Bcl-2VEGF-protein and mRNA in the retina of the rats in each group at different time points after model making (P 0.05 ~ 4 weeks). The expression of Nrf2HO-1VEGFmRNA in DM group was higher than that in NC group. The expression of Nrf2HO-1VEGFmRNA in the retina of the rats in each group was significantly higher than that in the NC group. The expression of Nrf2HO-1Bcl 2 was significantly lower in P0.05THQ group than that in DM group, and the difference was statistically significant (both P0.05 and 12 weeks after treatment), the expression of Nrf2HO-1Bcl-2VEGF in DM group was higher than that in NC group, and the expression of Nrf2HO-1Bcl-2VEGF in DM group was significantly lower than that in DM group. There was significant difference in the expression of Nrf2HO-1Bcl 2 between the two groups. The expression of VEGF in DM group was significantly lower than that in DM group. The expression of VEGF in DM group was significantly lower than that in DM group at week 12 and week 4. The expression of VEGF in DM group was significantly higher than that in DM group at 4 weeks, and the expression of HO-1 in DM group was lower than that in DM group at 4 weeks, and the expression of HO-1Bcl-2 in DM group was significantly lower than that in DM group. There were significant differences in the expression of Nrf2HO-1Bcl 2 between the two groups (P 0.05). Conclusion the expression of Nrf2HO-1Bcl-2 in the retina of DM rats is protected by tBHQ, which can promote the expression of Nrf2HO-1Bcl-2 in the retina of DM rats. TBHQ may protect the retina of DM rats through Nrf2/HO-1/VEGF and Nrf2/Bcl-2 pathway, such as decreasing the expression of VEGF, damaging the retina tissue, reducing apoptosis and inhibiting the proliferation of retinal blood vessels.
【作者单位】：西南医科大学附属医院眼科;四川省人民医院眼科;大理白族自治州人民医院眼科;第三军医大学附属第二医院眼科;
【基金】：四川省科学技术厅资助项目(编号:2015SZ0086,LZ-LY-37)~~
【分类号】：R587.2;R774.1

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