莱菔硫烷对糖尿病视网膜病变Nrf2通路和NLRP3炎性小体的影响

Effect of sulforaphane on Nrf2 pathway and NLRP3 inflammatory bodies in diabetic retinopathy

目的:探讨莱菔硫烷对糖尿病视网膜病变Nrf2通路和NLRP3炎性小体的影响。方法:将SD大鼠分为空白对照组(blank control group,BC组)、模型组(model group,M组)、阳性对照组(positive contorl group,PC组)、SFN低剂量组(SFN low dose group,SLD组)和SFN高剂量组(SFN high dose group,SHD组)。M组、PC组、SLD组和SHD组采用高脂饲料联合STZ法造模,造模成功后,SLD组给予SFN 25mg/kg腹腔注射,SHD组给予SFN 50mg/kg腹腔注射,PC组给予羟苯磺酸钙溶液1.0/kg腹腔灌注,BC组和M组给予等量生理盐水腹腔注射,连续干预6周。观察大鼠视网膜组织形态、炎性因子和氧化还原指标水平、Nrf2-2、HO-1mRNA水平和蛋白表达水平。结果:M组大鼠视网膜细胞排列疏松,内外核层抛离紊乱,细胞水肿明显;PC组、SLD和SHD组视网膜结构较M组清晰,细胞水肿明显减少;SHD组改善较PC组和SLD组更为明显。M组MDA、IL-1β、TNF-α、NLRP3蛋白、ASC蛋白、caspase-1蛋白、TXNIP蛋白水平显著高于BC组(P<0.05),SOD、Nrf2mRNA、HO-1mRNA、Nrf2蛋白、HO-1蛋白水平低于BC组(P<0.05);PC组和SLD组MDA、IL-1β、TNF-α、NLRP3蛋白、ASC蛋白、caspase-1蛋白、TXNIP蛋白水平低于M组(P<0.05),SOD、Nrf2mRNA、HO-1mRNA、Nrf2蛋白、HO-1蛋白水平高于M组(P<0.05);SHD组MDA、IL-1β、TNF-α、NLRP3蛋白、ASC蛋白、caspase-1蛋白、TXNIP蛋白水平低于M组、PC组和SLD组(P<0.05),SOD、Nrf2mRNA、HO-1mRNA、Nrf2蛋白、HO-1蛋白水平高于M组、PC组和SLD组(P<0.05)。结论:SFN可能通过激活Nrf2信号通路,降低氧化应激和TXNIP水平,从而降低NLRP3信号通路的激活,从而对DR产生保护作用。

Objective:To investigate the effect of sulforaphane on Nrf2 pathway and NLRP3 inflammatory body in diabetic retinopathy.Methods:SD rats were divided into blank control group(BC group),model group(M group),positive control group(PC group),SFN low dose group(SLD group)and SFN high dose group(SHD group).In SLD,SHD,PC,BC and M group,SFN25mg/kg,SFN 50mg/kg,Calcium dobesilate solution 1.0/kg,normal saline and normal saline were given intraperitoneal injection,respectively.The morphology of retina,levels of inflammatory factors and redox index,nrf2-2,HO-1 mRNA and protein expression were observed.Results:In group M,the arrangement of retinal cells was loose,the internal and external nuclear layers were scattered disorderly,and the cell edema was obvious.In group PC,SLD and SHD,the retinal structure was clear and the cell edema was obviously reduced,and in group SHD,the improvement was more obvious than that in group PC and SLD.MDA,IL-1β,TNF-α,NLRP3 protein,ASC protein,caspase-1 protein,TXNIP protein levels in group M were significantly higher than those in group BC(P<0.05),and SOD,Nrf2 mRNA,HO-1mRNA,Nrf2 protein,HO-1 protein levels in group M were lower than those in group BC(P<0.05).The levels of MDA,IL-1β,TNF-α,NLRP3,ASC,caspase-1 and TXNIP in PC group and SLD group were lower than those in M group(P<0.05),and the levels of SOD,Nrf2 mRNA,HO-1mRNA,Nrf2 protein and HO-1 protein were higher than those in M group(P<0.05).The levels of MDA,IL-1β,TNF-α,NLRP3 protein,ASC protein,caspase-1 protein,TXNIP protein in SHD group were lower than those in M group,PC group and SLD group(P<0.05),and the levels of SOD,Nrf2 mRNA,HO-1mrna,Nrf2 protein,HO-1 protein in SHD group were higher than those in M group,PC group and SLD group(P<0.05).Conclusion:SFN may reduce the level of oxidative stress and TXNIP by activating Nrf2 signaling pathway,reduce the activation of NLRP3 signaling pathway,and protect DR.