MiR-1-3p在VEGF/Notch信号通路介导青光眼模型视网膜神经节细胞凋亡的机制研究  

Mechanism of MiR-1-3p in apoptosis of retinal ganglion cells in glaucoma rats via VEGF/Notch signaling pathway

作　　者：顾丹 罗娜 GU Dan;LUO Na(Department of Ophthalmology,the 903rd Hospital of Jiangyou City,Sichuan Province,Jiangyou 621700,China)

机构地区：[1]四川省江油市九〇三医院眼科,四川江油621700

出　　处：《河北医科大学学报》2025年第2期202-207,共6页Journal of Hebei Medical University

基　　金：四川省医学(青年创新)科研课题立项(S21207)。

摘　　要：目的探讨miR-1-3p对青光眼大鼠视网膜神经节细胞凋亡的影响以及其调节机制。方法将36只1月龄SPF级SD大鼠进行适应性饲喂1周,之后采用随机数字表法均分为正常组、模型组和miR-1-3p拮抗组(每组12只)。其中,正常组的大鼠未进行任何处理。模型组的大鼠构建青光眼模型,术后每日腹腔注射无菌生理盐水0.2 mL,连续干预7 d。miR-1-3p拮抗组的大鼠构建青光眼模型,术后每日腹腔注射3μmol/L的miR-1-3p拮抗剂0.2 mL,连续干预7 d。在青光眼模型造模结束及最后一次治疗干预时测量大鼠眼压。利用蛋白质免疫印迹法测定各组大鼠视网膜组织中Notch1和Notch2的水平。利用酶联免疫吸附试验(enzyme-linked immunosorbent assay,ELISA)检测血管内皮生长因子(vascular endothelial growth factor,VEGF)含量。利用qRT-PCR试验测定各组大鼠视网膜组织中VEGF、Notch1和Notch2 mRNA表达水平。利用原位末端标记法(TdT-mediated biotinylated-dUTP nick end labeling,TUNEL)检测试剂盒测定细胞凋亡情况。结果造模结束时模型组和miR-1-3p拮抗组的眼压均显著高于正常组(P<0.05)。最后一次治疗干预结束时,miR-1-3p拮抗组的眼压显著低于模型组(P<0.05)。模型组视网膜组织中VEGF水平与Notch1和Notch2蛋白表达水平显著高于正常组(P<0.05)。与模型组相比,miR-1-3p拮抗组视网膜组织中VEGF水平与Notch1和Notch2蛋白表达水平显著降低(P<0.05)。模型组视网膜组织中VEGF、Notch1和Notch2 mRNA表达水平和细胞凋亡水平显著高于正常组(P<0.05)。与模型组相比,miR-1-3p拮抗组视网膜组织中VEGF、Notch1和Notch2 mRNA表达水平和细胞凋亡水平显著降低(P<0.05)。结论MiR-1-3p通过促进VEGF/Notch信号通路促进青光眼大鼠视网膜神经节细胞凋亡,是治疗青光眼的潜在分子靶标。Objective To investigate the effect of miR-1-3p on apoptosis of retinal ganglion cells in glaucoma rats and its regulatory mechanism.Methods Thirty-six one-month-old SPF SD rats were given adaptive feeding for one week,and then divided into normal group(n=12),model group(n=12)and miR-1-3p antagonistic group(n=12)by random number table method.The normal group did not receive any treatment.The rats in the model group were used to construct glaucoma model,and 0.2 mL sterile saline was injected intraperitoneally every day for 7 days.The rats in the miR-1-3p antagonist group were used to construct glaucoma model,and 0.2 mL of 3μmol/L miR-1-3p antagonist was injected intraperitoneally every day after surgery for continuous intervention for 7 days.Intraocular pressure(IOP)was measured at the end of glaucoma modeling and the last intervention.The levels of Notch1 and Notch2 in the retinal tissues of each group were measured by Western blotting,and vascular endothelial growth factor(VEGF)content was detected by enzyme-linked immunosorbent assay(ELISA).The mRNA expression levels of VEGF,Notch1 and Notch2 in retinal tissues of rats were determined by qRT-PCR,and apoptosis was determined by TdT-mediated biotinylated-dUTP nick end labeling(TUNEL)assay.Results At the end of modeling,the IOP of model group and miR-1-3p antagonist group was significantly higher than that of normal group(P<0.05).At the end of the last intervention,the IOP of miR-1-3p antagonist group was significantly lower than that of model group(P<0.05).VEGF levels and Notch1 and Notch2 protein expression levels in retinal tissues of model group were significantly higher than those of normal group(P<0.05).Compared with the model group,VEGF levels and Notch1 and Notch2 protein expression levels in retinal tissues of miR-1-3p antagonistic group were significantly decreased(P<0.05).The mRNA expression levels of VEGF,Notch1,Notch2 and apoptosis levels in retinal tissues in model group were significantly higher than those in normal group(P<0.05).Compared with the

关 键 词：青光眼 视网膜神经节细胞 细胞凋亡 

分 类 号：R775.2[医药卫生—眼科]