Purpose

To explore the role of nucleotide-binding oligomerization domain-like receptors (NLRs) family caspase-activation and the recruitment domain containing 4 (NLRC4) inflammasome in retinal ganglion cell (RGC) injury induced by an acute glaucoma mouse model.

Method

A mouse model of acute ocular hypertension, which can lead to retinal ischemia-reperfusion (I/R) injury, was established. The expression level of NLRC4 was detected by polymerase chain reaction and western blotting. Localized expression of NLRC4 was detected by examining immunofluorescence in eyeball sections. Intravitreal adeno-associated virus 2(AAV2) administration was used to knockdown retinal Nlrc4. Fluoro-Gold labeled RGCs and TdT-mediated dUTP nick end labeling were used to evaluate the survival and apoptosis of RGCs. Tlr4^−/− mice were utilized to explore whether NLRC4 inflammasome is influenced by Toll-like receptor4 (TLR4).

Results

NLRC4, expressed in RGCs and microglial cells, was actively involved in mouse retinal I/R injury. Knockdown of Nlrc4 using an AAV2 vector caused an obvious reduction in the generation of IL-1β led by the rapidly elevated intraocular pressure, and thereby improved the RGC survival. In addition, activation of the NLRC4 inflammasome could influence the phosphorylation of p38 and Jun N-terminal kinase, which was largely dependent on TLR4 signaling.

Conclusion

Our study demonstrated the role of NLRC4 inflammasome in promoting RGC damage in mouse retinal I/R injury. Inhibition of NLRC4 might be leveraged as a potential therapeutic target in glaucomatous retinopathy.

中文翻译：

---

NLRC4炎性小体参与促进急性青光眼小鼠模型中视网膜神经节细胞死亡

目的

探讨核苷酸结合寡聚域样受体（NLRs）家族胱天蛋白酶激活和包含4（NLRC4）炎性小体的募集域在急性青光眼小鼠模型引起的视网膜神经节细胞（RGC）损伤中的作用。

方法

建立了可导致视网膜缺血再灌注（I / R）损伤的急性高眼压小鼠模型。通过聚合酶链反应和蛋白质印迹检测NLRC4的表达水平。通过检查眼球切片中的免疫荧光来检测NLRC4的局部表达。玻璃体内腺相关病毒2（AAV2）管理被用来击倒视网膜Nlrc4。荧光金标记的RGC和TdT介导的dUTP缺口末端标记用于评估RGC的存活和凋亡。Tlr4 ^-/-小鼠用于研究NLRC4炎性小体是否受Toll样受体4（TLR4）影响。

结果

在RGC和小胶质细胞中表达的NLRC4积极参与小鼠视网膜I / R损伤。使用AAV2载体对Nlrc4进行基因敲除导致眼压迅速升高导致的IL-1β产生明显减少，从而提高了RGC的存活率。此外，NLRC4炎性小体的激活可能会影响p38和Jun N端激酶的磷酸化，这在很大程度上取决于TLR4信号传导。

结论

我们的研究证明了NLRC4炎性小体在促进小鼠视网膜I / R损伤中RGC损伤中的作用。NLRC4的抑制可能被用作青光眼性视网膜病的潜在治疗靶标。