RAGE抑制剂TTP488降低糖尿病性视网膜病变小鼠模型中的视网膜炎症的疗效OA
Efficacy of RAGE Inhibitor TTP488 in Reducing Retinal Inflamma-tion in Diabetic Retinopathy Mice Model
目的 深入探索晚期糖基化终末产物受体抑制剂 TTP488 降低视网膜炎症的疗效及其作用机制.方法 30 只雄性 C57BL/6J 小鼠随机分入 3 组(n=10):对照组、1 型糖尿病(type I diabetes mellitus,T1DM)模型组和 T1DM+TTP488 组.建立链脲佐菌素(streptozotocin,STZ)诱导的 T1DM 小鼠模型.免疫荧光染色测定小鼠视网膜组织中小胶质细胞标记物(Iba1)阳性细胞活化程度;ELISA 测定其中炎症因子 TNF-α、IL-1β、IL-6 的水平.体外细胞实验分组为细胞对照组、LPS 组、LPS+TTP488 组、LPS+shR-NA-P65 组和 LPS+shRNA-NT 组.采用脂多糖(lipopolysaccharide,LPS)诱导建立小鼠视网膜色素上皮原代细胞炎症模型.腺病毒载体介导敲低 P65.RT-qPCR 法测定细胞内VEGF-D mRNA 的表达水平.蛋白质印迹法测定视网膜组织中和体外细胞内磷酸化 NF-κB P65(p-P65)/P65 和 VEGF-D 的表达水平.结果 与T1DM 模型组比较,T1DM+TTP488 组小鼠视网膜组织中 Iba1 阳性细胞数量、细胞分支数和细胞分支连接点数(每视野个数)均减少(P<0.05);TNF-α、IL-1β、IL-6 水平均降低(P<0.05);p-P65/P65 和VEGF-D 相对表达水平均降低(P<0.05).与 LPS 组比较,LPS+TTP488 组和 LPS+shRNA-P65 组 VEGF-D mRNA 相对表达水平均降低(P<0.05);p-P65 和 VEGF-D 相对表达水平均下调(P<0.05).结论 TTP488 通过抑制 NF-κB 的活化下调 VEGF-D 的表达,可降低糖尿病性视网膜病变小鼠模型中的视网膜炎症和组织结构病变.
Objective To explore the efficacy and mechanisms of the receptor for advanced glycation end products inhibitor TTP488 in reducing retinal inflammation.Methods Thirty male C57BL/6J mice were randomly divided into 3 groups(n=10 each):control group,type I diabe-tes mellitus(T1DM)model group,and T1DM+TTP488 group.A T1DM mice model induced by streptozotocin(STZ)was established.Immunofluorescent staining targeting microglial cell markers(Iba1)was used to determine the activation degree of Iba1-positive cells in the retinal tissues of mice;and ELISA was used to measure the levels of inflammatory factors TNF-α,IL-1β and IL-6 in them.mRPE cells was divided into the following 5 groups for the further in vitro cell experiments:control group,LPS group,LPS+TTP488 group,LPS+shRNA-P65 group,LPS+shRNA-NT group.Lipopolysaccharide(LPS)was used to establish the primary mouse retinal pigment epithelial cells inflammatory model.Adenovirus vector was used to mediate the knockdown of P65.The expres-sion level of VEGF-D mRNA in cells was determined by RT-qPCR.The expression levels of phospho-rylated NF-κB P65(p-P65)/P65 and VEGF-D in the retinal tissues of mice as well as in in vitro cells were determined by Western blotting.Results Compared with those in the T1DM model group,the number of Iba1 positive cell branches and the number of Iba1 positive cell branch con-nection points(per field)in the T1DM+TTP488 group were decreased(P<0.05).The levels of TNF-α,IL-1β,and IL-6 were decreased(P<0.05).The relative expression levels of p-P65/P65 and VEGF-D were decreased(P<0.05).Compared with those in the LPS group,the relative ex-pression level of VEGF-D mRNA in the LPS+TTP488 group and the LPS+shRNA-P65 group was de-creased(P<0.05).The relative expression levels of p-P65 and VEGF-D were down-regulated(P<0.05).Conclusion TTP488 reduced the expression of VEGF-D by inhibiting the activation of NF-κB,thereby alleviating retinal inflammation and tissue structural abnormalities in the diabetic retinopathic mice model.
王岚;卢怡;邹云春
首都医科大学附属北京安贞医院南充医院南充市中心医院 四川省 南充市,637000四川省信息中心 成都市,610000首都医科大学附属北京安贞医院南充医院南充市中心医院 四川省 南充市,637000
医药卫生
糖尿病性视网膜病变炎症晚期糖基化终末产物晚期糖基化终末产物受体TTP488血管内皮生长因子
diabetic retinopathyinflammationadvanced glycation end productsreceptor for advanced glycation end productsTTP488vascular endothelial growth factor
《医学分子生物学杂志》 2026 (3)
294-301,8
四川省基层卫生发展研究中心资助课题(No.SWFZ23-C-95) This work was supported by a grant from the Sichuan Provincial Center for Primary Health Care Development(No.SWFZ23-C-95).
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