miR-184在葡萄膜炎发病过程中对白细胞介素-18和TLR-8表达的调控作用机制

目的:检测miR-1、miR184与多巴胺转运体在多巴胺细胞系SH-SY5Y内的表述,研究miR、miR-184在SH-SY5Y细胞系中对DAT基因表述的作用与其体制。方法:使用microRNA.org、miRBase、TargetScan.等网络数据库预估与文献检索相融合的模式,选择可以作用于DAT基因的候补miRNAs,并化学合成候补miRNAs模拟物转染到SH-SY5Y细胞内。依次转染后12h与24h后,提炼NC-mimics组、NC-inhibitor组、miRNAs mimics组与miRNAs inhibitor组细胞总RNA与蛋白;使用实时荧光定量RT-PCR检测各组细胞内的候补miRNA、DATmRNA表述情况;使用Western Blot检测各组细胞内DAT蛋白表述水平。结果:(1)通过生物信息理论检测融合检索资料选择出的候补miRANs是miR-1、miR-184;(2)miRNAs表述水平:miR-1 mimics 12h组与miR-1 mimics 24h组的miR-1表述量明显超过对照组(Ps<0.001),miR-1 inhibitor 12h组、miR-1 inhibitor 24h组合对照组比较没有明显差异(Ps>0.05);miR-184 mimics24h处理组中miR-184的表述量和对照组比较都有明显差异(P<0.01),miR-184 inhibitor 12h、miR-184 mimics12h与miR-184 inhibitor 24h处理组与对照组比较并无明显差异(Ps>0.05);(3)DATmRNA表述水平:miR-1 mimics 12h处理组内DAT mRNA的表述量和对照组比较有明显差异(P<0.05),miR-1 mimics 24h、miR-1 inhibitor 12h组与miR-1inhibitor 24h组DAT mRNA的表述量和对照组比较都缺乏明显差异(Ps>0.05),miR-184 mimics 12h组、miR-184 inhibitor 12h组与miR-184 inhibitor 24h组DAT mRNA的表述量和对照组比较都缺乏明显差异(Ps<0.05)。结论miR-1、miR-184参加SH-SY5Y细胞内DAT基因表述的控制,过表达的miR-1与miR-184都能压抑SH-SY5Y细胞内的DAT蛋白的表述。

Homer1a reduces inflammatory response after retinal ischemia/reperfusion injury

Elevated intraocular pressure(IOP)is one of the causes of retinal ischemia/reperfusion injury,which results in NRP3 inflammasome activation and leads to visual damage.Homerla is repo rted to play a protective role in neuroinflammation in the cerebrum.However,the effects of Homerla on NLRP3inflammasomes in retinal ischemia/reperfusion injury caused by elevated IOP remain unknown.In our study,animal models we re constructed using C57BL/6J and Homer1^(flox/-)/Homerla^(+/-)/Nestin-Cre^(+/-)mice with elevated IOP-induced retinal ischemia/repe rfusion injury.For in vitro expe riments,the oxygen-glucose deprivation/repe rfusion injury model was constructed with M uller cells.We found that Homerla ove rexpression amelio rated the decreases in retinal thickness and Muller cell viability after ischemia/reperfusion injury.Furthermore,Homerla knockdown promoted NF-κB P65^(Ser536)activation via caspase-8,NF-κB P65 nuclear translocation,NLRP3 inflammasome formation,and the production and processing of interleukin-1βand inte rleukin-18.The opposite results we re observed with Homerla ove rexpression.Finally,the combined administration of Homerla protein and JSH-23 significantly inhibited the reduction in retinal thickness in Homer1^(flox/-)Homer1a^(+/-)/Nestin-Cre^(+/-)mice and apoptosis in M uller cells after ischemia/reperfusion injury.Taken together,these studies demonstrate that Homer1a exerts protective effects on retinal tissue and M uller cells via the caspase-8/NF-KB P65/NLRP3 pathway after I/R injury.