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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

崔乐乐  王云松  单明华  周云涛  田卫   

  1. 063000河北省唐山工人医院眼科
  • 收稿日期:2015-12-11 出版日期:2016-05-25 发布日期:2016-06-22
  • 通讯作者: 王云松,Email:wyunsong@163.com E-mail: wyunsong@163.com
  • 基金资助:
    河北省省级科技计划项目(13277783D)

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

CUI Le-le, WANG Yun-song, SHAN Ming-hua, ZHOU Yun-tao, TIAN Wei   

  1. Department of Ophthalmology, Tanshan Gongren Hospital, Tangshan 063000, China
  • Received:2015-12-11 Online:2016-05-25 Published:2016-06-22
  • Contact: WANG Yun-song, Email: wyunsong@163.com E-mail: wyunsong@163.com

摘要: 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5 mmol/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、ATF4 mRNA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。

关键词: 糖尿病视网膜病变;喷他佐辛;Mü, ller细胞;血管内皮生长因子;转录激活因子4

Abstract: bjective 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.

Key words: diabetic retinopathy, Pentazocine, Müller cells, VEGF, ATF4