[1]徐卫星,李勇,徐军.维生素B12对糖尿病大鼠视网膜神经细胞凋亡的影响[J].眼科新进展,2017,37(12):1114-1118.[doi:10.13389/j.cnki.rao.2017.0281]
　XU Wei-Xing,LI Yong,XU Jun.Effects of vitamin B12 on apoptosis of retinal ganglion cells of diabetic rat[J].Recent Advances in Ophthalmology,2017,37(12):1114-1118.[doi:10.13389/j.cnki.rao.2017.0281]
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维生素B12对糖尿病大鼠视网膜神经细胞凋亡的影响/HTML
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《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
37卷
期数:
2017年12期
页码:
1114-1118
栏目:
实验研究
出版日期:
2017-12-05

文章信息/Info

Title:
Effects of vitamin B12 on apoptosis of retinal ganglion cells of diabetic rat
作者:
徐卫星; 李勇; 徐军
121001　辽宁省锦州市，锦州医科大学附属第一医院眼科
Author(s):
XU Wei-Xing; LI Yong; XU Jun
Department of Ophthalmology，the First Affiliated Hospital of Jinzhou Medical University，Jinzhou 121001，Liaoning Province，China
关键词:
维生素B12; 糖尿病视网膜病变; 氧化应激; Caspase-3; 凋亡
Keywords:
vitamin B12; diabetic retinopathy; oxidative stress; Caspase-3; apoptosis
分类号:
R774
DOI:
10.13389/j.cnki.rao.2017.0281
文献标志码:
A
摘要:
目的　观察维生素B12对糖尿病大鼠视网膜神经细胞凋亡的影响并分析其作用机制。方法　取清洁级SD大鼠36只，随机分为正常对照组（NC组）、糖尿病模型组（DM组）、维生素B12治疗组（VitB12组），每组12只。DM组和VitB12组腹腔注射链脲佐菌素建立糖尿病大鼠模型。确定造模成功后，每天对VitB12组进行维生素B12（100 μg·kg-1）灌胃，持续8周，NC组和DM组用同体积生理盐水替代。停止灌胃2周后，于第10周末将所有大鼠处死，取颈静脉血，用于检测血清中丙二醛含量及超氧化物歧化酶的活性；取视网膜组织，用于制备视网膜石蜡切片，采用TUNEL法进行视网膜凋亡细胞原位标记，计算视网膜神经节细胞凋亡指数（apoptosis index,AI）；采用免疫组织化学法检测视网膜组织Caspase-3蛋白的表达。结果　三组大鼠血清丙二醛含量比较，差异有统计学意义（F=471.061，P=0.000），DM组、VitB12组丙二醛含量高于NC组（均为P＜0.01），DM组高于VitB12组（P＜0.01）；三组间血清超氧化物歧化酶活性差异有统计学意义（F=356.923，P=0.000），DM组、VitB12组超氧化物歧化酶活性低于NC组（均为P＜0.01），DM组低于VitB12组（P＜0.01）。三组大鼠视网膜神经节细胞层、内核层和外核层均可见凋亡细胞，DM组和VitB12组凋亡细胞数较NC组明显增多，VitB12组凋亡细胞数较DM组有不同程度减少。三组视网膜神经节细胞AI值比较差异有统计学意义（F=14.926，P=0.000），DM组、VitB12组AI值均高于NC组（均为P＜0.01），DM组AI值高于VitB12组（P＜0.05）。显微镜观察发现三组大鼠视网膜神经节细胞层、内核层和外核层均可见Caspase-3蛋白表达，DM组和VitB12组Caspase-3蛋白表达均较NC组明显增多，VitB12组Caspase-3蛋白表达较DM组明显减少。三组视网膜Caspase-3光密度值比较差异有统计学意义（F=77.896，P=0.000），DM组、VitB12组视网膜Caspase-3光密度值均高于NC组（均为P＜0.01），DM组高于VitB12组（P＜0.01）。结论　维生素B12对糖尿病视网膜神经细胞有一定的保护作用，其机制可能是维生素B12通过抗氧化作用抑制Caspase-3的表达从而抑制神经细胞的凋亡。
Abstract:
Objective　To investigate the effects of vitamin B12 on apoptosis in retinal ganglion cells of rat with diabetes and its underlying mechanisms.Methods　Together 36 clean SD rats were randomly divided into 3 groups （n=12）：negative control （NC） group，diabetic mellitus （DM） group and vitamin B12 treatment （VitB12） group.Then a model diabetic rats was conducted by intraperitoneal administration of streptozotocin in rats of DM and VitB12 groups.After modeling successfully，100 μg·kg-1 vitamin B12 was given to rats in the VitB12 group by intragastric administration 1 time per day for 8 consecutive weeks，and an equivalent amount of normal saline was utilized for the animals of NC and DM groups in the same way.After 10 weeks，all the rats were sacrificed to collect jugular blood for detection of serum malondialdehyde （MDA） level and superoxide dismutase （SOD） activity；and the retinal tissue was also collected for preparation of paraffin sections for observing the apoptosis of retinal ganglion cells and counting the apoptotic index （AI） by TUNEL staining.Finally，the expression of Caspase-3 in the retina was detected by immunohistochemistry.Results　The levels of serum MDA in the DM and VitB12 groups were both higher than that in the NC group （all P<0.01），and DM group was higher than VitB12 group （P<0.01）；and there was significant difference among the three groups（F=471.061，P=0.000）.The activity of SOD in the DM and VitB12 groups were both lower than that in the NC group （all P<0.01），and DM group was lower than VitB12 group （P<0.01）；and there was significant difference in the three groups （F=356.923，P=0.000）.Apoptotic cells were observed in retinal ganglion cell layer，inner and outer nuclear layer in three groups，and the number of apoptotic cells in the DM and VitB12 groups was significantly larger than that in the NC group，but the number of apoptotic cells in VitB12 group was decreased to some extent compared with DM group.The AI of retinal ganglion cells in the DM and VitB12 groups were both higher than that in the NC group （all P<0.01），and DM group was higher than VitB12 group （P<0.05）；and there was significant difference among the three groups （F=14.926，P=0.000）.Microscopic examination showed Caspase-3 protein expression was appeared in the retinal ganglion cell layer，the inner and the outer nuclear layer in the three groups.The Caspase-3 protein expression in DM and VitB12 groups was significantly increased compared with NC group，and VitB12 group was significantly lower than DM group.Moreover，the optical density of Caspase-3 in the DM and VitB12 groups was higher than that in the NC group （all P<0.01），and DM group was higher than VitB12 group （P<0.01），and there was significant difference among the three groups （F=77.896，P=0.000）.Conclusion　Vitamin B12 has certain protective effect on retinal ganglion cell of diabetic rats，of which the mechanism may be its antioxidant stress can inhibit the expression of Caspase-3，thereby suppressing the apoptosis of retinal ganglion cells.

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备注/Memo

备注/Memo:
辽宁省科技厅联合基金项目（编号：2015020351）
更新日期/Last Update: 2017-12-08