Baclofen Protects Primary Rat Retinal Ganglion Cells from Chemical Hypoxia-Induced Apoptosis Through the Akt and PERK Pathways.
Baclofen Protects Primary Rat Retinal Ganglion Cells from Chemical Hypoxia-Induced Apoptosis Through the Akt and PERK Pathways.
复制标题
巴氯芬通过 Akt 和 PERK 途径保护原代大鼠视网膜神经节细胞免受化学缺氧诱导的细胞凋亡
DOI：
10.3389/fncel.2016.00255
复制
发表时间：
2016
影响因子：
5.3
通讯作者：
Gao F
中科院分区：
文献类型：
--
作者：
Fu P;Wu Q;Hu J;Li T;Gao F
视网膜神经节细胞（RGCs）消耗大量能量将光信息转化为神经元信号，这使得它们极易受到缺氧损伤。本研究旨在探讨GABAB受体激动剂巴氯芬对RGCs缺氧诱导的细胞凋亡的潜在保护作用。采用氯化钴（CoCl2）模拟缺氧。将原代大鼠RGCs在加或不加巴氯芬的情况下进行CoCl2处理，并使用RNA干扰技术敲低原代RGCs中的GABAB2基因。采用细胞活力、末端脱氧核苷酸转移酶介导的dUTP缺口末端标记（TUNEL）、Hoechst染色和流式细胞术评估RGCs的活力和凋亡情况。western blotting检测cleaved caspase-3、bcl-2、bax、Akt、phospho-Akt、protein kinase RNA (PKR) like ER kinase （PERK）、phospho-PERK、eIF2α、phospho-eIF2α、ATF-4和CCAAT/增强子结合蛋白同源蛋白（CHOP）的表达。采用实时定量聚合酶链反应（qRT-PCR）检测GABAB2 mRNA的表达。我们的研究表明，CoCl2显著诱导RGC细胞凋亡，而巴氯芬逆转了这一作用。cocl2诱导的Akt活性降低也被巴氯芬逆转。巴氯芬可阻止CoCl2诱导的PERK通路激活和CHOP表达增加。抑制GABAB2的下调和Akt通路的失活降低了巴氯芬对cocl2处理的RGCs的保护作用。综上所述，这些结果表明，巴氯芬通过增加Akt活性、抑制PERK通路和CHOP激活来保护RGCs免受cocl2诱导的凋亡。
Retinal ganglion cells (RGCs) consume large quantities of energy to convert light information into a neuronal signal, which makes them highly susceptible to hypoxic injury. This study aimed to investigate the potential protection by baclofen, a GABAB receptor agonist of RGCs against hypoxia-induced apoptosis. Cobalt chloride (CoCl2) was applied to mimic hypoxia. Primary rat RGCs were subjected to CoCl2 with or without baclofen treatment, and RNA interference techniques were used to knock down the GABAB2 gene in the primary RGCs. The viability and apoptosis of RGCs were assessed using cell viability and terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) assays, Hoechst staining, and flow cytometry. The expression of cleaved caspase-3, bcl-2, bax, Akt, phospho-Akt, protein kinase RNA (PKR)-like ER kinase (PERK), phospho-PERK, eIF2α, phospho-eIF2α, ATF-4 and CCAAT/enhancer-binding protein homologous protein (CHOP) were measured using western blotting. GABAB2 mRNA expression was determined using quantitative real-time polymerase chain reaction (qRT-PCR) analysis. Our study revealed that CoCl2 significantly induced RGC apoptosis and that baclofen reversed these effects. CoCl2-induced reduction of Akt activity was also reversed by baclofen. Baclofen prevented the activation of the PERK pathway and the increase in CHOP expression induced by CoCl2. Knockdown of GABAB2 and the inactivation of the Akt pathway by inhibitors reduced the protective effect of baclofen on CoCl2-treated RGCs. Taken together, these results demonstrate that baclofen protects RGCs from CoCl2-induced apoptosis by increasing Akt activity and by suppressing the PERK pathway and CHOP activation.
登录
查看更多内容
影响因子：
5.6
作者：
Chen JH;Kuo HC;Lee KF;Tsai TH
通讯作者：
Tsai TH
影响因子：
8
作者：
Cascio, S.;Bartella, V.;Surmacz, E.
通讯作者：
Surmacz, E.
影响因子：
4.4
作者：
Kaur, Charanjit;Sivakumar, Viswanathan;Ling, Eng-Ang
通讯作者：
Ling, Eng-Ang
影响因子：
64.8
作者：
Calfon, M;Zeng, HQ;Ron, D
通讯作者：
Ron, D
影响因子：
3.4
作者：
Koulen, P;Malitschek, B;Brandstätter, JH
通讯作者：
Brandstätter, JH