喷他佐辛对高糖环境下体外培养视网膜Müller细胞的保护作用

Protective effects of pentazocine on retinal Müller cells under high-glucose conditions

目的探索喷他佐辛对高糖环境下体外培养大鼠视网膜Müller细胞的保护作用及其作用机制。设计实验研究。研究对象体外培养的SD大鼠视网膜Müller细胞。方法体外培养的SD大鼠视网膜Müller细胞分三组:正常对照组(葡萄糖5mmol/L),高糖组(葡萄糖30 mmol/L),高糖+喷他佐辛组(葡萄糖30 mmol/L+喷他佐辛3 umol/L),分别在培养6、12、24、72小时以MTT法检测细胞活性;RT-PCR法检测Müller细胞血管内皮生长因子(vascular endothelial growth factor,VEGF)和转录激活因子4(activating transcription factor 4,ATF4)mRNA的表达;蛋白印迹法检测VEGF和ATF4蛋白的表达。主要指标细胞活性,VEGF和ATF4 mRNA和蛋白的表达。结果高糖作用72小时,高糖组与对照组相比细胞活性下降(23.3±0.1)%(P<0.05),喷他佐辛处理后可抑制由于高糖作用造成的Müller细胞活性降低,且与对照组无统计学差异。与对照组相比,高糖组VEGF、ATF4mRNA和蛋白表达量均显著增加(P<0.05),喷他佐辛处理后可降低VEGF、ATF4 mRNA和蛋白的表达与对照组无显著性差异(P>0.05)。结论喷他佐辛可提高高糖作用下体外培养视网膜Müller细胞的活性,减少高糖作用下视网膜Müller细胞VEGF、ATF4 mRNA和蛋白的表达,具有神经保护作用。

Objective To study the protective effects and mechanism of Pentazocine（PTZ） on retinal Müller cells under high-glucose conditions. Design Experimental study. Participants The cultivated retinal Müller cells of SD rats. Methods The cultivated retinal Müller cells of SD rats were divided into three groups, control group with normal level of glucose（NG,5.5 mmol/L）, high glucose group（HG, 30 mmol/L）, Pentazocine treatment group（HG＋PTZ, HG in the presence of PTZ 3 umol/L）. MTT is used to detect cellular activity at 6, 12, 24 and 72 h cultivated. Vascular endothelial growth factor（VEGF） and activating transcription factor 4（ATF4） mRNA expression by the Müller cells in three experimental groups were examined by RT-PCR, production of VEGF and ATF4 was measured by Western blot. Main Outcome Measures Cellular activity, expression of mRNAs and proteins. Results After 72 h, high glucose exposed, the HG group cells viabilities decreased significantly compared with NG（P〈0.05）, and the cells vitality of HG group were decreased by（23.3±0.1）% compared with NG group; HG＋PTZ group cells activities was higher than the HG group（P〈0.05）. After the retinal Müller cells were exposed to HG（30 mmol/L）, compared with NG, the expressions of VEGF, ATF4 mRNA and protein were up regulated（P〈0.05）. In addition, Pentazocine reduced HG-induced the expression of VEGF, ATF4 mRNA and protein, there was no statistical difference between NG and HG＋PTZ groups（P〈0.05）. Conclusion Pentazocine significantly improve the cellular acticvity and inhibited HG-induced VEGF and ATF4 mRNA and protein expression in retinal Müller cells.