腺苷A_(2A)受体(A_(2A)R)拮抗剂ZM241385对视网膜脱离动物模型视网膜光感受器细胞的保护作用

Protective effect of adenosine A_(2A)receptor antagonist ZM241385 on retinal photoreceptor cells in mice with retinal detachment

目的探讨腺苷A_(2A)受体(A_(2A)R)拮抗剂ZM241385对视网膜脱离(RD)动物模型视网膜光感受器细胞的保护作用。方法选取健康雄性SPF级C57BL/6J小鼠48只作为研究对象。将实验小鼠随机分为对照+二甲基亚砜(DMSO)组、对照+ZM241385组、RD+DMSO组、RD+ZM241385组,每组12只。RD模型建立后,实验小鼠立即腹腔注射ZM241385(3 mg·kg^(-1),剂量不超过0.2 mL)或同体积DMSO,连续给药3 d。采用免疫荧光染色检测视网膜中Müller细胞谷氨酰胺合成酶(GS)的表达,Western blot检测单核细胞趋化蛋白(MCP)-1、肿瘤坏死因子(TNF)-α、Cleaved caspase 3和B细胞淋巴瘤抗体(Bcl)-2的表达。结果免疫荧光染色结果显示:RD+DMSO组小鼠视网膜Müller细胞GS荧光强度较对照+DMSO组显著降低,而RD+ZM241385组小鼠视网膜Müller细胞GS荧光强度较RD+DMSO组显著升高。RD+DMSO组小鼠视网膜MCP-1、TNF-α蛋白表达水平较对照+DMSO组均显著升高,差异均有统计学意义(均为P<0.001),而RD+ZM241385组小鼠视网膜MCP-1、TNF-α蛋白表达水平较RD+DMSO组均显著降低,差异均有统计学意义(均为P<0.001)。与对照+DMSO组相比,RD+DMSO组小鼠视网膜Cleaved caspase 3蛋白表达水平显著升高,Bcl-2蛋白表达水平显著下降,差异均有统计学意义(均为P<0.001)。而与RD+DMSO组相比,RD+ZM241385组小鼠视网膜Cleaved caspase 3蛋白表达水平显著下降,Bcl-2蛋白表达水平显著升高,差异均有统计学意义(均为P<0.001)。结论A_(2A)R拮抗剂ZM241385通过促进RD小鼠视网膜Müller细胞GS表达,抑制炎症细胞因子MCP-1、TNF-α的表达,缓解光感受器细胞凋亡。

Objective To explore the protective effect of adenosine A_(2A)receptor(A_(2A)R)antagonist ZM241385 on retinal photoreceptor cells in mice with retinal detachment(RD).Methods Forty-eight healthy SPF-grade C57BL/6J male mice were selected and randomly divided into the control+dimethyl sulfoxide(DMSO)group,control+ZM241385 group,RD+DMSO group,and RD+ZM241385 group,with 12 mice in each group.After RD modeling,mice were intraperitoneally injected with the A_(2A)R antagonist ZM241385(3 mg·kg^(-1))at a dose of not more than 0.2 mL or an equivalent volume of DMSO continuously for 3 days.Glutamine synthase(GS)expression in retinal Müller cells of mice in each group was evaluated with immunofluorescence.The expression levels of monocyte chemoattractant protein(MCP)-1,tumor necrosis factor(TNF)-α,Cleaved caspase 3,and B-cell lymphoma(Bcl)-2 were detected with Western blot.Results Immunofluorescence results showed that the GS fluorescence intensity of retinal Müller cells in the RD+DMSO group was significantly lower than that in the control+DMSO group and that the GS fluorescence intensity of retinal Müller cells in the RD+ZM241385 group was significantly higher than that in the RD+DMSO group.The expression levels of MCP-1 and TNF-αproteins in the RD+DMSO group were significantly higher than those in the control+DMSO group(both P<0.001),while the expression levels of MCP-1 and TNF-α proteins in the RD+ZM241385 group were significantly lower than those in the RD+DMSO group(both P<0.001).Compared with the control+DMSO group,the expression level of Cleaved caspase 3 protein in the RD+DMSO group was significantly increased,and the expression level of Bcl-2 protein was significantly decreased(both P<0.001).Compared with the RD+DMSO group,the expression level of Cleaved caspase 3 protein in the RD+ZM241385 group was significantly decreased,and the expression level of Bcl-2 protein was significantly increased(both P<0.001).Conclusion AR antagonist ZM241385 alleviates the apoptosis of photorecep-tor cells in RD mice by promoting the expression of GS in retinal Müller cells and inhibiting the expression of inflammatory cytokines MCP-1 and TNF-α.