沙格列汀调控miR-23b-3p/EGR1轴对高糖诱导足细胞损伤的影响及机制研究

Study on effect and mechanism of saxagliptin regulating miR-23b-3p/EGR1 axis on podocyte injury induced by high glucose

目的 探讨沙格列汀通过调控微小RNA(micro RNA,miR)-23b-3p/早期生长反应因子(early growth response factor,EGR)1轴对高糖诱导足细胞损伤的影响。方法 将人足细胞分为对照组、高糖组、高糖+沙格列汀组、高糖+沙格列汀+anti-miR-对照组以及高糖+沙格列汀+anti-miR-23b-3p组。分别检测细胞miR-23b-3p、EGR1信使RNA(messenger RNA,mRNA)表达、增殖、凋亡、活性氧(reactive oxygen species,ROS)、丙二醛(malondialdehyde,MDA)、超氧化物歧化酶(superoxide dismutase,SOD)、白介素(interleukin,IL)-1β、IL-6、肌间线蛋白、裂孔隔膜蛋白(Podocin、Nephrin)、B细胞淋巴瘤-2相关X蛋白(B cell lymphoma-2 related X protein,Bax)、半胱氨酸蛋白酶(Caspase)-3以及EGR1表达,并验证miR-23b-3p与EGR1的靶向关系。结果 与对照组相比,高糖组的miR-23b-3p表达水平、增殖率、SOD、Podocin以及Nephrin均降低,EGR1表达、凋亡率、IL-1β、IL-6、MDA、ROS、Caspase-3、Bax以及肌间线蛋白均升高(P <0.05);与高糖组相比,高糖+沙格列汀组miR-23b-3p表达、增殖率、SOD、Podocin以及Nephrin均升高,EGR1表达、凋亡率、IL-1β、IL-6、MDA、ROS、caspase-3、Bax以及肌间线蛋白均降低(P <0.05);下调miR-23b-3p可减弱沙格列汀对足细胞损伤的改善(P <0.05);miR-23b-3p靶向负调控EGR1表达。结论 沙格列汀可能通过上调miR-23b-3p负调控EGR1,减轻高糖诱导的足细胞凋亡和损伤。

Objective To investigate the effect of saxagliptin on podocyte injury induced by high glucose by regulating the microRNA(miR)-23b-3p/early growth response factor(EGR)1 axis.Methods Human podocytes were divided into control group,high glucose group,high glucose+saxagliptin group,high glucose+saxagliptin+anti-miR-control group and high glucose+saxagliptin+anti-miR-23b-3p group.The expression of miR-23b-3p,EGR1 messenger RNA(mRNA),proliferation,apoptosis,reactive oxygen species(ROS),malondialdehyde(MDA),superoxide dismutase(SOD),interleukin(IL)-1β,IL-6,desmin,split pore membrane proteins(Podocin,Nephrin),B cell lymphoma-2 related X protein(Bax),Caspase-3 and EGR1 protein were detected,the targeting relationship between miR-23b-3p and EGR1 was verified.Results Compared with control group,the expression level of miR-23b-3p,proliferation rate,SOD,Podocin and Nephrin in high glucose group were obviously decreased,while the expression of EGR1 mRNA and protein,apoptosis rate,IL-1β,IL-6,MDA,ROS,Caspase-3,Bax and desmin were increased(P<0.05).Compared with high glucose group,the expression level of miR-23b-3p,proliferation rate,SOD,Podocin and Nephrin in high glucose+saxagliptin group were obviously increased,while the expression of EGR1 mRNA and protein,apoptosis rate,IL-1β,IL-6,MDA,ROS,Caspase-3,Bax and desmin were decreased(P<0.05).Down-regulation of miR-23b-3p could attenuate the improvement of saxagliptin on podocyte injury(P<0.05).MiR-23b-3p negatively regulated the expression of EGR1.Conclusion Saxagliptin may reduce high glucose-induced apoptosis and injury of podocytes by up-regulating miR-23b-3p and negatively regulating EGR1.