Neuroprotective Effects of C-Type Natriuretic Peptide on Rat Retinal Ganglion Cells
Neuroprotective Effects of C-Type Natriuretic Peptide on Rat Retinal Ganglion Cells
复制标题
C型利钠肽对大鼠视网膜神经节细胞的神经保护作用
DOI：
10.1167/iovs.09-5049
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发表时间：
2010-07-01
影响因子：
4.4
通讯作者：
Pang, Iok-Hou
中科院分区：
文献类型：
--
作者：
Ma, Jia;Yu, Wenhan;Pang, Iok-Hou
目的。探讨c型利钠肽（CNP）对大鼠视网膜神经节细胞（RGCs）的潜在神经保护作用。培养的成年大鼠视网膜细胞用载体、CNP或心房利钠肽（ANP）处理，然后进行细胞毒性损伤（谷氨酸、TNF α或营养因子的退出）。通过计数每孔中thy -1阳性细胞来分析RGC存活。为了进行体内评价，将n -甲基- d -天冬氨酸（NMDA）加或不加CNP注入大鼠眼内。在注射后的不同时间点，分析视网膜横截面，观察视网膜层的厚度变化，并通过计数甲酚紫标记或tunel阳性细胞来评估视网膜平载。采用定量逆转录-聚合酶链反应（qRT-PCR）分析视网膜中利钠肽受体- b （NPRB）和凋亡相关基因Bcl-xL、BAX、mu-calpain的表达。在50和500 nM时，CNP对谷氨酸损伤和营养因子戒断诱导的RGC体外死亡有显著保护作用（P < 0.05）。两种肽均未显著影响TNF α诱导的细胞毒性。玻璃体内注射NMDA（20纳米摩尔）显著（P < 0.05）降低内层丛状层（IPL）厚度，诱导细胞损失，增加RGC层tunel阳性细胞数量，上调Bcl-xL、BAX和mu-calpain的表达。同时注射CNP （4.5 nmol, 10 mu g）可显著减轻上述影响（P < 0.05）。注射CNP后，其神经保护作用可维持至14天。CNP在体外和体内均能保护大鼠RGCs免受兴奋性氨基酸等损伤引起的凋亡损伤。（中国眼科杂志，2010;51:3544-3553）DOI: 10.1167/iovs.09-5049
PURPOSE. To evaluate the potential neuroprotective effects of C-type natriuretic peptide (CNP) on rat retinal ganglion cells (RGCs).METHODS. Cultured adult rat retinal cells were treated with vehicle, CNP, or atrial natriuretic peptide (ANP), followed by cytotoxic insults (glutamate, TNF alpha, or withdrawal of trophic factor). RGC survival was analyzed by counting Thy-1-positive cells in each well. For in vivo evaluation, N-methyl-D-aspartate (NMDA) with or without CNP was injected intravitreally into rat eyes. At various time points after injection, retinal cross-sections were analyzed for thickness changes in the retinal layers, and retinal flat mounts were assessed by counting cresyl violet-labeled or TUNEL-positive cells. Expressions of natriuretic peptide receptor-B (NPRB) and apoptosis-related genes in retina, including Bcl-xL, BAX, and mu-calpain, were analyzed by quantitative reverse transcription-polymerase chain reaction (qRT-PCR).RESULTS. At 50 and 500 nM, CNP, but not ANP, significantly (P < 0.05) protected against glutamate-insult and trophic factor withdrawal-induced RGC death in vitro. Neither peptide significantly affected TNF alpha-induced cytotoxicity. Intravitreal injection of NMDA (20 nanomoles) significantly (P < 0.05) decreased the thickness of the inner plexiform layer (IPL), induced cell loss, increased the number of TUNEL-positive cells in the RGC layer, and upregulated the expression of Bcl-xL, BAX, and mu-calpain. All these effects were significantly (P < 0.05) alleviated by concomitant injection of CNP (4.5 nmol, 10 mu g). The neuroprotective effects of CNP were maintained up to 14 days after CNP injection.CONCLUSIONS. CNP protects rat RGCs against the apoptotic damage induced by insults such as excitatory amino acid, both in vitro and in vivo. (Invest Ophthalmol Vis Sci. 2010;51:3544-3553) DOI: 10.1167/iovs.09-5049