Maresin1通过抑制NOD样受体家族蛋白3炎症小体活化改善糖尿病大鼠视网膜炎症反应

Maresin1 improves retinal inflammation by inhibiting NOD-like receptor protein 3 inflammasome activation in diabetic rats

目的探讨Maresin1(MaR1)通过抑制NOD样受体家族蛋白3(NLRP3)炎症小体介导的炎症反应来保护大鼠糖尿病视网膜病变(DR)。方法SPF级SD大鼠,建立糖尿病模型。按照随机数字表法分为4组:正常(Con)组、糖尿病(DM)组、MaR1组、MaR1+NLRP3激活剂尼日利亚菌素组(Nigericin)组。成模后,MaR1组4 ng/g MaR1腹腔注射给药,每周2次,Con组和DM组给予等量PBS注射。Nigericin组1 mg/kg Nigericin每天1次腹腔注射和4 ng/g MaR1每周2次腹腔注射联合给药。HE染色观察组织病理改变。免疫组化检测神经胶质纤维酸性蛋白(GFAP)表达。ELISA测定视网膜白介素(IL)-18、IL-1β、IL-6水平。Western blot检测视网膜IL-18、IL-1β、IL-6、NLRP3、凋亡相关斑点样蛋白(ASC)、天冬酰胺特异酶半胱氨酸蛋白酶(Caspase-1)表达。结果DM组神经节细胞(RGC)数量明显少于Con组,GFAP表达高于Con组,IL-18、IL-1β、IL-6、NLRP3、ASC、Caspase-1表达水平高于Con组,差异有统计学意义(P<0.05)。MaR1组RGC数量多于DM组,GFAP表达低于DM组,IL-18、IL-1β、IL-6、NLRP3、ASC、Caspase-1表达水平低于DM组,差异有统计学意义(P<0.01)。然而在MaR1组基础上给予NLRP3激活剂Nigericin后,MaR1的保护作用被逆转。结论MaR1能够明显抑制DR炎症反应,这可能与抑制NLRP3炎症小体激活有关。

Objective To investigate the protective effect of Maresin1(MaR1)against diabetic retinopathy(DR)by inhibiting NOD-like receptor protein 3(NLRP3)inflammasomes mediated inflammatory response in rats.Methods SPF SD rats were be conditioned to set up diabetes model.They were divided into 4 groups according to the random number table method:Control(Con)group,Diabetes Mellitus(DM)group,MaR1 group and MaR1 combined with NLRP3 activator Nigericin(Nigericin)group.After modeling,MaR1 group was i.p injected with 4 ng/g MaR1 twice a week,and Con and DM group were used to isometric PBS.Nigericin group was given a Nigericin injection of 1 mg/kg with a combined intraperitoneal injection of 4 ng/g MaR1 twice a week.Histopathological alters were saw by HE.Variation trend of glial fibrillary acidic protein(GFAP)was found by immunohistochemistry.The level of retinal Interleukin(IL)-18,IL-1βand IL-6 were assessed through ELISA.Variation trend of retinal IL-18,IL-1β,IL-6,NLRP3,apoptosis-associated speck-like protein(ASC)and cysteine aspartate-specific proteinase(Caspase-1)signaling proteins were found by Western blot.Results The number of retinal ganglion cell(RGC)in the DM group was less than that in the Con group,the expression of GFAP was higher than that in the Con group,and the expression levels of IL-18,IL-1β,IL-6,NLRP3,ASC,and Caspase-1 were higher than those in the Con group,with statistical significances(P<0.05).The number of RGC in the MaR1 group was higher than that in the DM group,the expression of GFAP was lower than that in the DM group,and the expression levels of IL-18,IL-1β,IL-6,NLRP3,ASC,and Caspase-1 were lower than those in the DM group,and the differences were statistically significant(P<0.01).However,the protective effect of MaR1 was reversed when nigericin,a NLRP3 activator,was administered on a MaR1 basis.Conclusion MaR1 could restrain the phlogosis of DR,which could be related to the suppression of NLRP3 activation.