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目的:研究高糖环境对原代培养新生7天SD乳鼠视网膜Mü ller细胞谷氨酸转运合成系统的影响及其可能机制。方法:新生7天SD乳鼠视网膜Mü ller细胞原代培养并模拟高糖环境构建乳鼠视网膜mü ller细胞体外高糖环境模型。处理分为3组:对照组,高糖组,高糖+白藜芦醇干预组。培养时间为24 h,通过western blot等检测方法,对照观察各组Müller细胞谷氨酸转运体(GLAST)、谷氨酰胺合成酶(GS)的表达情况。结果:模拟高糖环境可以造成新生SD乳鼠视网膜Müller细胞谷氨酸转运体(GLAST)表达的降低(0.225 fold VS control,P<0.05),并导致其表达的谷氨酰胺合成酶(GS)表达水平的显著降低(0.653 fold VScontrol,P<0.05);而干预药物白藜芦醇作用后可明显逆转新生SD乳鼠Mü ller细胞谷氨酸转运体(GLAST)(1.133 fold VS H Ggroup,P<0.05)、谷氨酰胺合成酶(GS)(1.720 fold VS H G group,P<0.05)等蛋白的表达水平。结论:模拟高糖环境可以影响视网膜Müller细胞谷氨酸转运体(GLAST)、谷氨酰胺合成酶的表达,其结局可能导致视神经细胞因谷氨酸堆积而导致的兴奋性毒性,白藜芦醇能提高Müller细胞谷氨酸转运体(GLAST)、谷氨酰胺合成酶表达,从而保护视神经细胞。
AIM: To investigate the effects of high glucose on the glutamate transport system of retinal Müller cell cultured on primary cultured neonatal SD rats for 7 days and its possible mechanism. Methods: Primary cultured retinal Mü ller cells of SD neonatal rats were cultured and simulated in high glucose for 7 days to construct a high glucose environment model of neonatal retinal mü ller cells in vitro. The treatment was divided into 3 groups: control group, high glucose group, high glucose + resveratrol intervention group. The culture time was 24 h. The expression of glutamate transporter (GLAST) and glutamine synthetase (GS) in Müller cells of each group were observed by western blot and other methods. RESULTS: Simulated high glucose environment led to a decrease of glutamate transporter (GLAST) expression in retinal Müller cells (0.225 fold VS control, P <0.05), leading to a decrease in glutamine synthetase (GS) (0.653 fold VS control, P <0.05). However, resveratrol significantly reversed the GLAST (1.133 fold VS H Ggroup, P <0.05), and glutamine synthetase (GS) (1.720 fold VS HG group, P <0.05). CONCLUSION: Simulated high glucose environment may affect the expression of glutamate transporter (GLAST) and glutamine synthetase in retinal Müller cells. The outcome may result in the excitotoxicity of optic nerve cells due to accumulation of glutamate. Resveratrol Can enhance Müller cells glutamate transporter (GLAST), glutamine synthetase expression, thereby protecting optic nerve cells.