本文選題：維生素B + 糖尿病視網(wǎng)膜病變�。� 參考：《眼科新進展》2017年12期

【摘要】：目的觀察維生素B12對糖尿病大鼠視網(wǎng)膜神經(jīng)細胞凋亡的影響并分析其作用機制。方法取清潔級SD大鼠36只,隨機分為正常對照組(NC組)、糖尿病模型組(DM組)、維生素B12治療組(Vit B12組),每組12只。DM組和Vit B12組腹腔注射鏈脲佐菌素建立糖尿病大鼠模型。確定造模成功后,每天對Vit B12組進行維生素B12(100μg·kg~(-1))灌胃,持續(xù)8周,NC組和DM組用同體積生理鹽水替代。停止灌胃2周后,于第10周末將所有大鼠處死,取頸靜脈血,用于檢測血清中丙二醛含量及超氧化物歧化酶的活性;取視網(wǎng)膜組織,用于制備視網(wǎng)膜石蠟切片,采用TUNEL法進行視網(wǎng)膜凋亡細胞原位標記,計算視網(wǎng)膜神經(jīng)節(jié)細胞凋亡指數(shù)(apoptosis index,AI);采用免疫組織化學法檢測視網(wǎng)膜組織Caspase-3蛋白的表達。結果三組大鼠血清丙二醛含量比較,差異有統(tǒng)計學意義(F=471.061,P=0.000),DM組、Vit B12組丙二醛含量高于NC組(均為P0.01),DM組高于VitB 12組(P0.01);三組間血清超氧化物歧化酶活性差異有統(tǒng)計學意義(F=356.923,P=0.000),DM組、VitB 12組超氧化物歧化酶活性低于NC組(均為P0.01),DM組低于VitB 12組(P0.01)。三組大鼠視網(wǎng)膜神經(jīng)節(jié)細胞層、內核層和外核層均可見凋亡細胞,DM組和VitB 12組凋亡細胞數(shù)較NC組明顯增多,VitB 12組凋亡細胞數(shù)較DM組有不同程度減少。三組視網(wǎng)膜神經(jīng)節(jié)細胞AI值比較差異有統(tǒng)計學意義(F=14.926,P=0.000),DM組、VitB 12組AI值均高于NC組(均為P0.01),DM組AI值高于VitB 12組(P0.05)。顯微鏡觀察發(fā)現(xiàn)三組大鼠視網(wǎng)膜神經(jīng)節(jié)細胞層、內核層和外核層均可見Caspase-3蛋白表達,DM組和VitB 12組Caspase-3蛋白表達均較NC組明顯增多,VitB 12組Caspase-3蛋白表達較DM組明顯減少。三組視網(wǎng)膜Caspase-3光密度值比較差異有統(tǒng)計學意義(F=77.896,P=0.000),DM組、VitB 12組視網(wǎng)膜Caspase-3光密度值均高于NC組(均為P0.01),DM組高于VitB 12組(P0.01)。結論維生素B12對糖尿病視網(wǎng)膜神經(jīng)細胞有一定的保護作用,其機制可能是維生素B12通過抗氧化作用抑制Caspase-3的表達從而抑制神經(jīng)細胞的凋亡。
[Abstract]:Objective to observe the effect of vitamin B 12 on apoptosis of retinal nerve cells in diabetic rats and analyze its mechanism. Methods Thirty-six SD rats of clean grade were randomly divided into normal control group (NC group), diabetic model group (DM group), vitamin B12 treatment group (Vit B12) group (n = 12) and streptozotocin (Vit B 12) group (n = 12). The diabetic rats were induced by intraperitoneal injection of streptozotocin (STZ) in each group. After the model was established successfully, Vit B12 group was given vitamin B12 100 渭 g / kg / day intragastric perfusion for 8 weeks, the NC group and the DM group were replaced by normal saline of the same volume. After two weeks of gastric perfusion, all the rats were killed at the end of the 10th week, the jugular vein blood was taken to detect the malondialdehyde content in serum and the activity of superoxide dismutase, and the retina tissue was taken for the preparation of paraffin sections of the retina. The apoptosis index of retinal ganglion cells was calculated by TUNEL method, and the expression of Caspase-3 protein in retinal tissue was detected by immunohistochemical method. Results the content of malondialdehyde in serum of the three groups was compared. The content of malondialdehyde (MDA) in Vit B12 group was higher than that in NC group (P 0.01) and the activity of serum superoxide dismutase in DM group was higher than that in VitB 12 group (P 0.01) and the activity of serum superoxide dismutase in DM group was lower than that in Vit B12 group. It was lower in NC group (P 0.01) than in VitB 12 group (P 0.01). The number of apoptotic cells in the retinal ganglion cell layer, nuclear layer and outer nuclear layer of the three groups were significantly increased in DM group and VitB 12 group compared with NC group. The number of apoptotic cells in Vit B12 group was lower than that in DM group. The AI of retinal ganglion cells in DM group was significantly higher than that in NC group (P 0.01) and the AI value of DM group was higher than that of VitB group (P 0.05). Microscopic observation showed that the expression of Caspase-3 protein in retinal ganglion cell layer, nuclear layer and outer nuclear layer was significantly increased in DM group and VitB 12 group compared with NC group. The expression of Caspase-3 protein in Vit B12 group was significantly lower than that in DM group. There were significant differences in Caspase-3 optical density between the three groups. The Caspase-3 optical density of retina in Vit B12 group was significantly higher than that in NC group (P 0.01) and that in VitB 12 group was higher than that in VitB 12 group (P 0.01). Conclusion Vitamin B12 has a protective effect on diabetic retinal nerve cells, and its mechanism may be that vitamin B12 inhibits the expression of Caspase-3 and thus inhibits neuronal apoptosis through antioxidant action.
【作者單位】： 錦州醫(yī)科大學附屬第一醫(yī)院眼科;
【基金】：遼寧省科技廳聯(lián)合基金項目(編號:2015020351)~~
【分類號】：R587.2;R774.1

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