[1]田敏,余曦,吕红彬.PFKFB3、S1P2在2型糖尿病大鼠视网膜中的表达及tBHQ的干预作用[J].眼科新进展,2018,38(2):131-135.[doi:10.13389/j.cnki.rao.2018.0028]
　TIAN Min,YU Xi,LV Hong-Bin.Expression of PFKFB3 and S1P2 in the retina of type 2 diabetic rats and the intervention effect of tBHQ[J].Recent Advances in Ophthalmology,2018,38(2):131-135.[doi:10.13389/j.cnki.rao.2018.0028]
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PFKFB3、S1P2在2型糖尿病大鼠视网膜中的表达及tBHQ的干预作用/HTML
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《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
38卷
期数:
2018年2期
页码:
131-135
栏目:
实验研究
出版日期:
2018-02-05

文章信息/Info

Title:
Expression of PFKFB3 and S1P2 in the retina of type 2 diabetic rats and the intervention effect of tBHQ
作者:
田敏; 余曦; 吕红彬
646000　四川省泸州市，西南医科大学附属医院眼科
Author(s):
TIAN Min; YU Xi; LV Hong-Bin
Department of Ophthalmology，the Affiliated Hospital of Southwest Medical University，Luzhou 646000，Sichuan Province，China
关键词:
6-磷酸果糖激酶-2/果糖双磷酸酶-2同工酶3; 1-磷酸鞘氨醇受体2; 叔丁基对苯二酚; 糖尿病; 视网膜
Keywords:
PFKFB3; S1P2; tBHQ; diabetes; retina
分类号:
R774.1
DOI:
10.13389/j.cnki.rao.2018.0028
文献标志码:
A
摘要:
目的　研究6-磷酸果糖激酶-2/果糖双磷酸酶-2同工酶3（6-phosphofructo-2-kinase/fructose-2，6-bisphosphatase-3，PFKFB3）、1-磷酸鞘氨醇受体2（sphingosine 1-phosphate receptor 2，S1P2）在2型糖尿病（diabetes mellitus，DM）大鼠视网膜中的表达，并探讨叔丁基对苯二酚（tertiary butylhydroquinone，tBHQ）与PFKFB3、S1P2的关系。方法　雄性Sprague Dawley大鼠60只，分为正常对照组（NC组）、DM组和tBHQ组。后两组诱导2型DM模型，其中tBHQ组于造模成功后7 d在高脂高糖饲料中添加10 g·L-1 tBHQ进行干预，DM组大鼠继续使用高脂高糖饲料喂养。于tBHQ干预后4周和12周各组大鼠心脏采血，检测血清空腹血糖（fasting plasma glucose，FPG）、空腹血清胰岛素（fasting serum insulin，FINs）含量。采用免疫组织化学法及实时荧光定量PCR（qRT-PCR）法定量检测各组大鼠视网膜中PFKFB3、S1P2、VEGF蛋白及mRNA的分布和表达。采用TUNEL法检测各组大鼠视网膜神经节细胞的凋亡指数。结果　三组大鼠在4周和12周的FPG、FINs总体比较差异均有统计学意义（均为P=0.000）。4周和12周时各组中均可见PFKFB3、S1P2、VEGF蛋白的阳性表达，且主要分布于视网膜神经节细胞层、内核层。免疫组织化学及qRT-PCR检测结果显示，各组大鼠在造模后不同时间点视网膜中PFKFB3、S1P2、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P<0.05）。4周和12周时，DM组PFKFB3、S1P2、VEGF的表达较NC组增加，差异均有统计学意义（均为P<0.05）；tBHQ组PFKFB3、S1P2、VEGF的表达较DM组降低，差异均有统计学意义（均为P<0.05）。与4周比较，12周时DM组PFKFB3、S1P2、VEGF的蛋白和mRNA表达均增加（均为P<0.05），tBHQ组S1P2蛋白和mRNA的表达较4周降低，差异均有统计学意义（均为P<0.05）。TUNEL法检测结果显示，4周和12周时DM组凋亡指数均较NC组增多，tBHQ组均较DM组减少（均为P<0.05）；与4周比较，12周时DM组凋亡指数增高，tBHQ组降低（均为P<0.05）。结论　PFKFB3、S1P2及VEGF共同参与2型DM大鼠视网膜的病理过程，其可能是通过PFKFB3/VEGF/S1P2信号途径起作用，并可能与视网膜神经节细胞的凋亡相关；tBHQ对2型DM大鼠视网膜有一定的保护作用。
Abstract:
Objective　To investigate the expression of 6-phosphofructo-2-kinase /fructose-2，6-bisphosphatase-3 (PFKFB3) and sphingosine 1-phosphate receptor 2 (S1P2) in the retina of rats with type 2 diabetes mellitus (DM) and to explore the correlation of tertiary butylhydroquinone (tBHQ) with PFKFB3 and S1P2.Methods　Together 60 male SD rats were divided into normal control group (NC group)，diabetes mellitus group (DM group) and tBHQ group.Type 2 DM model was induced in the latter two groups.The rats in tBHQ group were given 10 g·L-1 tBHQ in high-fat and high-sugar diet 1 week after successful modeling，while DM rats were fed with high-fat and high-sugar diet continuously.At 4 weeks and 12 weeks after tBHQ intervention，blood samples were taken from the hearts of rats in each group，and serum contents of fasting plasma glucose (FPG) and fasting serum insulin (FINs) were measured.Immunohistochemistry and qRT-PCR were taken to detect the distribution and expression of PFKFB3，S1P2 and vascular endothelial growth factor (VEGF) mRNA and protein in the retina，and TUNEL methods were used to detect apoptosis index of retinal ganglion cells of rats in each group.Results　There were significant difference in the FPG and FINs levels of the three groups at 4 weeks and 12 weeks (all P=0.000).Light microscopy test showed that positive expressions of PFKFB3，S1P2 and VEGF protein were found in all groups at 4 and 12 weeks，which were mainly located in the retinal ganglion cell layer and the inner nuclear layer.Immunohistochemistry and qRT-PCR showed that the difference in relative expressions of PFKFB3，S1P2，VEGF protein and mRNA in the retina at different times after modeling were statistically significant (all P<0.05).At 4 and 12 weeks，the expression levels of PFKFB3，S1P2 and VEGF in DM group were higher than those in NC group，but their expressions in tBHQ group were significantly downregulated when compared with DM group，and all differences were statistically significant(all P<0.05).Compared with 4 weeks，PFKFB3，S1P2 and VEGF mRNA and protein in DM group were overexpressed at 12 weeks，and the expression level of S1P2 in tBHQ group at 12 weeks was lower than that at 4 weeks，and the differences were statistically significant (all P<0.05).TUNEL assays showed that there was significant difference in the apoptotic index (AI) of retinal ganglion cells of rats in each group at different times after modeling (all P<0.05).DM group had higher AI than NC group (P<0.05)，and tBHQ group was lower than DM group (P<0.05)；Compared with 4 weeks，DM group had increased AI at 12 weeks while tBHQ group had decreased AI (both P<0.05).Conclusion　PFKFB3，S1P2 and VEGF are involved in the pathological process of the retina of type 2 DM rats，which may play a role through the PFKFB3 / VEGF / S1P2 signaling pathway and may be related to the apoptosis of retinal ganglion cells.And it is indicated that tBHQ has a protective effect on the retina of type 2 DM rats.

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备注/Memo

备注/Memo:
四川省教育厅基金资助项目（编号：17ZA0428）
更新日期/Last Update: 2018-02-02