低剂量氯喹介导的对n-甲基-d-天冬氨酸诱导的成年小鼠兴奋性毒性的神经保护作用

Q4 Medicine

中华眼底病杂志 Pub Date : 2020-04-25 DOI:10.3760/CMA.J.CN511434-20190903-00275

Yifan Liu, Yin Shen

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引用次数: 0

摘要

目的探讨不同浓度氯喹对n-甲基-d-天冬氨酸（NMDA）损伤小鼠RGC的保护作用及其可能机制。方法将54只健康雄性C57/BL6小鼠随机分为三组，每组18只。低剂量氯喹组小鼠腹膜内注射氯喹溶液，剂量为每天10mg/kg。高剂量氯喹组小鼠腹膜内注射剂量为100mg/kg的氯喹溶液，对照组小鼠腹腔内注射相同体积的PBS。在腹膜内注射后2天进行NMDA玻璃体内注射，将5nmoles NMDA注射到左眼中，并且将相同体积的PBS注射到右眼中作为对照。在NMDA注射后7天进行视网膜斑块的RGC染色，并计算存活的RGC的数量。在建模后9天和10天，用视力和视网膜电图评估RGC的电生理功能。NMDA注射后11天进行实时定量PCR和视网膜冷冻切片以及胶质纤维酸性蛋白（GFAP）免疫荧光染色，以评估视网膜的胶质细胞活化。通过单因素方差分析比较各组RGC的密度、视力和PhNR波振幅。结果NMDA注射后7天，低剂量氯喹组视网膜贴剂中RGC密度显著高于PBS对照组（F=54.41，P＜0.05），视力高于对照组，差异有统计学意义（F=9.10，P<0.05）。低剂量氯喹组PhNR波幅显著高于对照组（F=17.60，P<0.01）。炎症因子和GFAP阳性信号的mRNA水平也显著低于对照组（F=23.66，P<0.05），高剂量氯喹组的GFAP表达（F=110.20，P<0.01）和炎症因子mRNA水平（F=167.60，17.78；P<0.01）高于其他两组，差异有统计学意义。结论在NMDA损伤视网膜模型中，低剂量氯喹可显著提高RGC的存活率和生理功能，其作用机制可能与抑制胶质细胞活化和炎症反应有关。高剂量氯喹可加重RGC细胞凋亡。关键词：氯喹；视网膜神经节细胞；N-甲基天冬氨酸；毒性作用；动物实验

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Low-dose chloroquine mediated neuroprotection against n-methyl-d-aspartate induced excitotoxicity in adult mice

Objective To investigate the protective effects of different concentrations of chloroquine on RGC in n-methyl-d-aspartate (NMDA) injured mice and its possible mechanisms. Methods Fifty-four healthy male C57/BL6 mice were randomly divided into three groups, 18 in each group. The mice in low-dose chloroquine group were intraperitoneally injected with chloroquine solution at a dose of 10 mg/kg daily. Mice in high-dose chloroquine group were intraperitoneally injected with chloroquine solution at a dose of 100 mg/kg, and the mice in control group were intraperitoneally injected with the same volume of PBS. NMDA intravitreal injection was performed 2 days after intraperitoneal injection, 5 nmoles NMDA was injected into the left eye, and the same volume of PBS was injected into the right eye as a control. The RGC staining of retinal plaques were performed 7 days after NMDA injection, and the number of alive RGC was calculated. The visual acuity and electroretinogram were used to evaluate the electrophysiological functions of RGC at 9 and 10 days after modeling. Real-time quantitative PCR and retinal frozen sections and glial fibrillary acidic protein (GFAP) immunofluorescence staining were performed 11 days after NMDA injection to evaluate the glial activation of the retina. The density, visual acuity, and the amplitude of PhNR-wave of RGC between groups were compared by one-way analysis of variance. Results At 7 days after NMDA injection, the density of RGC in retinal patch of low-dose chloroquine group was significantly higher than that of intraperitoneal injection of PBS control group (F=54.41, P 0.05). The visual acuity was higher than control group, and the difference was statistically significant (F=9.10, P<0.05). The amplitude of PhNR-wave was significantly higher in low-dose chloroquine group than that of the control group (F=17.60, P<0.01). The mRNA level of inflammatory factor and GFAP positive signal was also significantly lower than that of the control group (F=23.66, P<0.05). The amplitude of PhNR-wave, the expression of GFAP (F=110.20, P<0.01) and the mRNA level of inflammatory factors (F=167.60, 17.78; P<0.01) in the high-dose chloroquine group were higher than the other two groups, and the differences were statistically significant. Conclusions In NMDA injury retinal model, low-dose chloroquine significantly increased the survival and physiological function of RGC, and the mechanism may be related to the inhibition of glial activation and inflammatory response. High-dose of chloroquine would aggravate the apoptosis of RGC. Key words: Chloroquine; Retinal ganglion cells; N-methylaspartate; Toxic actions; Animal experimentation

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来源期刊

中华眼底病杂志 Medicine-Ophthalmology

CiteScore

0.40

自引率

0.00%

发文量

5383

期刊介绍： Chinese Journal of Ocular Fundus Diseases is the only scientific journal in my country that focuses on reporting fundus diseases. Its purpose is to combine clinical and basic research, and to give equal importance to improvement and popularization. It comprehensively reflects the leading clinical and basic research results of fundus disease disciplines in my country; cultivates professional talents in fundus disease, promotes the development of fundus disease disciplines in my country; and promotes academic exchanges on fundus disease at home and abroad. The coverage includes clinical and basic research results of posterior segment diseases such as retina, uveal tract, vitreous body, visual pathway, and internal eye diseases related to systemic diseases. The readers are medical workers and researchers related to clinical and basic research of fundus diseases. According to the journal retrieval report of the Chinese Institute of Scientific and Technological Information, the comprehensive ranking impact factor and total citation frequency of the Chinese Journal of Ocular Fundus Diseases have been among the best in the disciplines of ophthalmology, otolaryngology, and ophthalmology in my country for many years. The papers published have been included in many important databases at home and abroad, such as Scopus, Peking University Core, and China Science Citation Database (CSCD).