siRNA靶向下调RhoA基因对慢性高眼压小鼠视功能的保护作用  

Protective Effect of RhoA Gene Down-Regulated by siRNA on Visual Function in Mice with Chronic Ocular Hypertension

作　　者：刘茜 刘长庚 李海军 董仰曾 张颖 LIU Qian;LIU Changgeng;LI Haijun;DONG Yangzeng;ZHANG Ying(Department of Ophthalmology,Henan Provincial People’s Hospital/Henan Provincial Eye Hospital/Henan Institute of Ophthalmology/People’s Hospital of Zhengzhou University,Zhengzhou 450000,China)

机构地区：[1]河南省人民医院/河南省立眼科医院/河南省眼科研究所/郑州大学人民医院眼科,河南郑州450000

出　　处：《河南医学研究》2023年第2期193-197,共5页Henan Medical Research

基　　金：河南省医学科技攻关计划省部共建项目(SBGJ2018083);河南省立眼科医院基础研究专项(21JCQN005)。

摘　　要：目的探究RNA干扰技术对青光眼视觉功能恢复的可行性。方法通过构建慢性高眼压小鼠模型并进行玻璃体腔注射RhoA siRNA(siRhoA)以干扰RhoA的表达,分别采用闪光视觉诱发电位(f-VEP)、定量反转录聚合酶链反应(qRT-PCR)和Western blot法对小鼠视觉功能、视网膜组织中RhoA基因和RhoA蛋白的表达情况进行检测。结果玻璃体腔注射后,siRhoA组小鼠患眼f-VEP P波、N波潜伏期和P波振幅逐渐恢复,相较NC siRNA和PBS组差异有统计学意义(P<0.05),NC siRNA和PBS组小鼠患眼f-VEP的P波和N波潜伏期延长,P波振幅降低(P<0.05)。此外,qRT-PCR和Western blot结果显示,经siRhoA处理后,慢性高眼压模型小鼠患眼视网膜组织RhoA基因和RhoA蛋白表达量与对侧眼相比差异无统计学意义(P>0.05);而NC siRNA和PBS组视网膜组织RhoA基因和RhoA蛋白表达量相较对侧眼差异有统计学意义(P<0.05)。结论本研究构建了慢性高眼压模型小鼠,并通过siRNA技术干扰了RhoA的活性,有效改善了青光眼模型小鼠视觉功能等,为基因途径治疗青光眼等疾病奠定了初步基础。Objective To explore the feasibility of RNA interference in the recovery of visual function of glaucoma.Methods A mice model of chronic high intraocular pressure was constructed and vitreous cavity injection of RhoA siRNA(siRhoA)was performed to interfere with the expression of RhoA.The flash visual evoked potential(f-VEP),quantitative reverse transcription polymerase chain reaction(qRT-PCR)and Western blot were used to examine the visual function,expression of RhoA gene and RhoA protein in the retinal tissues of mice,respectively.Results After intravitreal injection,the P-wave,N-wave latency and P-wave amplitude of f-VEP in the eyes of siRhoA group gradually recovered,which was statistically significant compared with NC siRNA group and PBS group(P<0.05),while the P-wave latency and N-wave amplitude of f-VEP in the eyes of NC siRNA group and PBS group were prolonged and the P-wave amplitude decreased(P<0.05).In addition,qRT-PCR and Western blot results showed that there was no statistically significant difference in the expression of RhoA gene and RhoA protein in the retina of chronic ocular hypertension model mice compared with the contralateral eyes(P>0.05),while the expression of RhoA gene and RhoA protein in retina of NC siRNA and PBS group was statistically significant compared with that of the contralateral eye(P<0.05).Conclusion This study constructed chronic ocular hypertension model mice,interfered with RhoA activity through siRNA technology,effectively improved the visual function of glaucoma model mice,and laid a preliminary foundation for gene therapy of glaucoma and other diseases.

关 键 词：青光眼 视网膜神经节细胞 RHOA SIRNA 

分 类 号：R770.4[医药卫生—眼科]