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表没食子儿茶素没食子酸酯对成年大鼠视网膜 N-甲基-D-天冬氨酸诱导的兴奋性毒性的神经保护作用。

Neuroprotective effect of epigallocatechin-3-gallate against N-methyl-D-aspartate-induced excitotoxicity in the adult rat retina.

机构信息

Beijing Ophthalmology and Visual Sciences Key Laboratory, Beijing Tongren Eye Center, Beijing Tongren Hospital, Capital Medical University, Beijing, China.

出版信息

Acta Ophthalmol. 2012 Dec;90(8):e609-15. doi: 10.1111/j.1755-3768.2012.02502.x. Epub 2012 Sep 13.

Abstract

PURPOSE

Epigallocatechin-3-gallate (EGCG), the major polyphenol of green tea, has been suggested to reduce glutamate excitotoxicity. We therefore investigated the potentially protective effects of EGCG against N-methyl-d-aspartate (NMDA)-induced excitotoxicity in the retina.

METHODS

Female Wistar rats (n = 171) were divided into a normal control group (n = 9); saline control group with intravitreal saline injections (n = 54); NMDA control group with an intravitreal NMDA injection and intraperitoneal saline injections (n = 54); and NMDA study group (n = 54) receiving an intravitreal NMDA injection plus intraperitoneal EGCG (25 mg/kg) injections. Starting at 2 days prior to the intravitreal NMDA injection, the intraperitoneal injections were performed daily for the whole study period. At 12 hr, 1, 2, 3 days, 1 and 2 weeks after the intravitreal NMDA injection, the animals were killed. We counted the neurons in the retinal ganglion cell layer (GCL) on histological sections, measured the thickness of Thy-1 immunoreactivity and assessed the expression of Thy-1 mRNA by real-time polymerase chain reaction.

RESULTS

At all time-points, GCL cell density, thickness of Thy-1 immunoreactivity and expression of Thy-1 mRNA were significantly (all p < 0.05) lower in the NMDA control group than in the NMDA study group, in which the parameters were significantly (all p < 0.05) lower than in the saline control group and the normal control group. In both groups with an intravitreal NMDA injection, GCL cell density, thickness of Thy-1 immunoreactivity and expression of Thy-1 mRNA decreased significantly with increasing follow-up time.

CONCLUSIONS

Intraperitoneal application of EGCG resulted in a significantly less marked NMDA-associated loss of retinal ganglion cells.

摘要

目的

表没食子儿茶素没食子酸酯(EGCG)是绿茶中的主要多酚,已被证明可减轻谷氨酸兴奋性毒性。因此,我们研究了 EGCG 对视网膜中 N-甲基-D-天冬氨酸(NMDA)诱导的兴奋性毒性的潜在保护作用。

方法

将雌性 Wistar 大鼠(n = 171)分为正常对照组(n = 9);生理盐水对照组,玻璃体内注射生理盐水(n = 54);NMDA 对照组,玻璃体内注射 NMDA 加腹腔注射生理盐水(n = 54);NMDA 研究组(n = 54),玻璃体内注射 NMDA 加腹腔注射 EGCG(25 mg/kg)。从玻璃体内 NMDA 注射前 2 天开始,每天进行腹腔注射,整个研究期间。在玻璃体内 NMDA 注射后 12 小时、1 天、2 天、3 天、1 周和 2 周时,处死动物。我们在组织切片上计数视网膜神经节细胞层(GCL)中的神经元,测量 Thy-1 免疫反应的厚度,并通过实时聚合酶链反应评估 Thy-1 mRNA 的表达。

结果

在所有时间点,NMDA 对照组的 GCL 细胞密度、Thy-1 免疫反应的厚度和 Thy-1 mRNA 的表达均明显低于 NMDA 研究组(均 p < 0.05),NMDA 研究组明显低于生理盐水对照组和正常对照组(均 p < 0.05)。在玻璃体内注射 NMDA 的两组中,GCL 细胞密度、Thy-1 免疫反应的厚度和 Thy-1 mRNA 的表达随着随访时间的增加而显著下降。

结论

腹腔内应用 EGCG 可显著减轻 NMDA 相关的视网膜神经节细胞丢失。

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