[1]张瑞雪,蒋文君,郭大东,等.玻璃体内注射原纤维蛋白-2 (FBN2)重组蛋白对FBN2缺陷型视网膜病变的影响[J].眼科新进展,2024,44(6):428-432.[doi:10.13389/j.cnki.rao.2024.0083]
 ZHANG Ruixue,JIANG Wenjun,GUO Dadong,et al.Effect of intravitreal injection of fibrillin-2 (FBN2) recombinant protein on FBN2-deficient retinopathy[J].Recent Advances in Ophthalmology,2024,44(6):428-432.[doi:10.13389/j.cnki.rao.2024.0083]
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玻璃体内注射原纤维蛋白-2 (FBN2)重组蛋白对FBN2缺陷型视网膜病变的影响/HTML
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《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
44卷
期数:
2024年6期
页码:
428-432
栏目:
实验研究
出版日期:
2024-06-03

文章信息/Info

Title:
Effect of intravitreal injection of fibrillin-2 (FBN2) recombinant protein on FBN2-deficient retinopathy
作者:
张瑞雪蒋文君郭大东石永伟毕宏生温莹
250014 山东省济南市,山东中医药大学(张瑞雪);250002 山东省济南市,山东省中西医结合眼病防治重点实验室,山东省高校中西医结合眼病防治技术(强化)重点实验室,山东省眼病防治研究院(蒋文君,郭大东,毕宏生);250002 山东省济南市,山东中医药大学附属眼科医院(石永伟,毕宏生,温莹)
Author(s):
ZHANG Ruixue1JIANG Wenjun2GUO Dadong2SHI Yongwei3BI Hongsheng23WEN Ying3
1.Shandong University of Traditional Chinese Medicine
2.Shandong Provincial Key Laboratory of Integrated Traditional Chinese and Western Medicine for Prevention and Therapy of Ocular Diseases,Key Laboratory of Integrated Traditional Chinese and Western Medicine for Prevention and Therapy of Ocular Diseases in Universities of Shandong,Shandong Academy of Eye Disease Prevention and Therapy
3.The Eye Hospital Affiliated to Shandong University of Traditional Chinese Medicine
关键词:
原纤维蛋白-2重组蛋白视网膜病变微原纤维相关糖蛋白-2I型胶原蛋白
Keywords:
fibrillin-2 recombinant protein retinopathy microfibril-associated glycoprotein 2 collagen I
分类号:
R774
DOI:
10.13389/j.cnki.rao.2024.0083
文献标志码:
A
摘要:
目的 探讨玻璃体内注射原纤维蛋白-2 (FBN2)重组蛋白对FBN2缺陷型视网膜病变的影响。
方法 取 SPF级C57BL/6J 小鼠32只,随机分为 4组:正常对照组、阴性对照组、FBN2敲减组、FBN2重组蛋白组,每组8只小鼠,取右眼作为实验眼。入组后,正常对照组小鼠不进行干预,阴性对照组小鼠玻璃体内注射3 μL 空载体(1 mg·L-1),FBN2敲减组及FBN2重组蛋白组小鼠玻璃体内注射3 μL腺相关病毒(AAV)(1 mg·L-1),4周后FBN2重组蛋白组小鼠玻璃体内注射3 μL FBN2重组蛋白(1 mg·L-1)。采用视网膜电图(ERG)视觉电生理仪和光学相干断层扫描(OCT)仪分别检测小鼠视网膜Rod-b、Max-a波振幅和视网膜结构变化;RT-PCR、Western blot检测小鼠视网膜中FBN2、微原纤维相关糖蛋白(MAGP-2)、I型胶原蛋白(COL1) mRNA和蛋白表达变化。
结果 ERG检测结果显示,与阴性对照组和正常对照组相比,FBN2敲减组和FBN2重组蛋白组小鼠视网膜ERG Rod-b、Max-a波振幅均变小(均为P<0.05);与FBN2敲减组相比,FBN2重组蛋白组视网膜ERG Rod-b、Max-a波振幅均明显增加(均为P<0.05)。OCT 检测结果显示,与FBN2敲减组相比,FBN2重组蛋白组小鼠视网膜色素上皮层组织结构恢复正常,光反射变规则。RT-PCR检测结果显示,与FBN2敲减组相比,FBN2重组蛋白组小鼠视网膜组织FBN2 mRNA表达明显升高,COL1、MAGP-2 mRNA表达均明显降低(均为P<0.05)。Western blot检测结果显示,与FBN2敲减组相比,FBN2重组蛋白组小鼠视网膜组织FBN2蛋白表达明显升高,COL1、MAGP-2蛋白表达均明显降低(均为P<0.05)。
结论 玻璃体内注射FBN2重组蛋白可以弥补FBN2缺陷型视网膜病变小鼠FBN2内源性缺失,通过调控COL1、MAGP-2表达可达到治疗疾病的作用。
Abstract:
Objective To investigate the effect of intravitreal injection of fibrillin-2 (FBN2) recombinant protein on FBN2-deficient retinopathy.
Methods Thirty-two SPF-grade C57BL/6J mice were randomly divided into 4 groups: normal control group, negative control group, FBN2 knockdown group, and FBN2 recombinant protein group, with 8 mice in each group. The right eyes were taken as the experimental eyes. Mice in the normal control group did not receive any intervention, mice in the negative control group were intravitreally injected with 3 μL empty vector (1 mg·L-1), and mice in the FBN2 knockdown group and FBN2 recombinant protein group were intravitreally injected with 3 μL adeno-associated virus (1 mg·L-1). After 4 weeks, mice in the FBN2 recombinant protein group were intravitreally injected with 3 μL FBN2 recombinant protein (1 mg·L-1). Then, electroretinogram (ERG) and optical coherence tomography (OCT) were used to measure the amplitude of Rod-b and Max-a waves and the changes in the retinal structure. Real-time quantitative polymerase chain reaction (RT-PCR) and Western blot were used to detect changes in FBN2, microfibril-associated glycoprotein 2 (MAGP-2), collagen I (COL1) mRNA and protein expression in the mouse retina.
Results The ERG findings showed that compared with the negative control group and normal control group, the amplitude of Rod-b and Max-a waves in the retina of mice in the FBN2 knockdown group and FBN2 recombinant protein group decreased (all P<0.05); compared with the FBN2 knockdown group, the amplitude of Rod-b and Max-a waves in the retina of mice in the FBN2 recombinant protein group significantly increased (both P<0.05).The OCT findings showed that compared with the FBN2 knockdown group, the structure of the retinal pigment epithelium and the light reflex in the FBN2 recombinant protein group became more regular. The RT-PCR detection results showed that compared with the FBN2 knockdown group, the expression of FBN2 mRNA in the retinal tissue of mice in the FBN2 recombinant protein group significantly increased, while the expression of COL1 and MAGP-2 mRNA significantly decreased (all P<0.05). Western blot assay results showed that compared with the FBN2 knockdown group, the expression of FBN2 protein in the retinal tissue of mice in the FBN2 recombinant protein group increased significantly, while the expression of COL1 and MAGP-2 proteins decreased significantly (all P<0.05).
Conclusion Intravitreal injection of FBN2 recombinant protein can compensate for the endogenous deficiency of FBN2 in mice with FBN2-deficient retinopathy and achieve therapeutic effects by regulating COL1 and MAGP-2 expression.

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备注/Memo

备注/Memo:
国家重点研发计划项目(编号:2021YFC2702103、2019YFC1710204、2021YFC2702100);山东省中西医结合防治疾病基金项目(编号:YXH2019ZXY001);山东省重点研发计划项目(编号:2019GSF108252)
更新日期/Last Update: 2024-06-05