小干扰RNA保护大鼠视网膜神经节细胞的实验研究

doi:10.13281/j.cnki.issn.1004-4469.2015.03.013

眼科 ›› 2015, Vol. 24 ›› Issue (3): 192-195.doi: 10.13281/j.cnki.issn.1004-4469.2015.03.013

小干扰RNA保护大鼠视网膜神经节细胞的实验研究

宋思玥马科赵秀丽

100730首都医科大学附属北京同仁医院国家药物临床试验机构北京市眼科研究所

收稿日期:2014-12-20 出版日期:2015-05-25 发布日期:2015-06-04
通讯作者: 赵秀丽，Email: xiulizhao@medmail.com.cn E-mail:xiulizhao@medmail.com.cn
基金资助:
十二五"重大新药创制"国家科技重大专项基金（2011ZX09302-007-05）

Neuroprotection of siRNA interference to the retinal ganglion cells in optic nerve crush model rat

SONG Si-yue, MA Ke, ZHAO Xiu-li

National Institute for Drug Clinical Trial,Beijing Institut of Ophthalmology, Beijing Tongren Hospital, Capital Medical University, Beijing 100730, China

Received:2014-12-20 Online:2015-05-25 Published:2015-06-04
Contact: ZHAO Xiu-li, Email: xiulizhao@medmail.com.cn E-mail:xiulizhao@medmail.com.cn

摘要/Abstract

摘要： 目的通过大鼠视神经夹伤模型，研究小干扰RNA（siRNA）对视网膜神经节细胞（RGC）的保护作用。设计实验研究。研究对象 SPF级SD大鼠54只。方法 54只SD大鼠随机分为A、B、C三组，每组18只。均选取右眼为实验眼，左眼为正常对照。在球后2 mm处用40 g压力微型视神经夹夹持视神经60 s，做视神经夹伤模型。建立模型后当天，A、B、C三组分别给予玻璃体注射10 μg、20 μg siRNA和生理盐水。视神经夹伤后10天，每组取6只大鼠用荧光金做逆行标记，14天时取标记后的大鼠双眼眼球标本做视网膜铺片并拍摄照片，RGC计数。计算RGC存活率（右眼RGC数/左眼RGC数×100%）。每组其余12只大鼠进一步用蛋白印迹法检测视网膜组织中caspase-3蛋白的表达水平。主要指标 RGC存活率，caspase-3蛋白的表达水平。结果 A、B、C组RGC存活率分别为53.63%±7.35%、57.86%±6.00%、45.00%±4.37%（F=7.11，P=0.029），其中A组与C组（P=0.025），B组和C组（P=0.002）之间均有显著性差异；A 组和B组之间无显著性差异（P=0.24）。A、B、C三组视网膜组织中Caspase-3蛋白与内参灰度比值分别为0.20±0.02、0.19±0.02、0.24±0.03（F=9.73，P=0.02）。其中A组与C组（P=0.005），B组和C组（P=0.001）之间均有显著性差异；A 组和B组之间无显著性差异（P=0.418）。结论小干扰RNA能有效保护大鼠视神经夹伤模型的RGC，提高RGC的存活率。

关键词: text-indent: 0px, display: inline !important, font: medium Simsun, white-space: normal, float: none, letter-spacing: normal, color: rgb(0, 0, 0), word-spacing: 0px, -webkit-text-stroke-width: 0px">视神经保护, 小干扰RNA, 视网膜神经节细胞

Abstract: Objective To investigate the neuroprotective effects of small interfering RNA (siRNA) on retinal ganglion cells (RGC) in optic nerve crush model rats. Design Experimental study. Participants fifty-four SD rats. Methods fifty-four SD rats were divided into 3 groups, group A, B, and C, 18 rats per group. The right eyes of all rats were crushed by micro-clipping with 40 g power for 60 s, the left eyes were not crushed as the controls. A and B group was given siRNA 10 μg/per eye and siRNA 20 μg/per eye, the group C was given normal saline intravitreal injectionon immediately after optic nerve crush. The experiment lasted for 14 days. Six rats in each group were retrograde labeled by injection of 3% fluorogold (FG) into both side of superior colliculus at 10 days after the crush. At the 14 day, the rats were sacrificed and stretched preparation of retina, RGCs were photographed and counted with mask method. The survival percentage of RGCs = RGCs density in right eye / RGCs density in left eye. Further the western blot was used to exam the expression level of caspase-3 protein in retinal tissue. Main Outcome Measures The survival percentage of RGC, the expression level of caspase-3 protein. Results The average survival percentage of RGCs in A, B and C group was 53.63%±7.35%, 57.86%±6.00%, 45.00%±4.37% respectively (F=7.11, P=0.029). A significant difference was found between group A and C(P=0.025), between group B and C(P=0.002) as well. The difference between group A and B was not significant (P=0.24). The expression level of caspase-3 protein in the A, B, C groups in the retinal tissue was 0.20±0.02, 0.19±0.02, 0.24±0.03 (F=9.73, P =0.02). A significant difference was found between group A and C(P=0.005), between group B and C(P=0.001) as well. The difference between group A and B was not significant(P=0.418). Conclusion siRNA shows the neuroprotective effects on RGCs in optic nerve crush model rats.

Key words: text-indent: 0px, display: inline !important, font: medium Simsun, white-space: normal, float: none, letter-spacing: normal, color: rgb(0,0,0), word-spacing: 0px, -webkit-text-stroke-width: 0px">neuroprotection, small interfering RNA, retinal ganglion cells

宋思玥马科赵秀丽. 小干扰RNA保护大鼠视网膜神经节细胞的实验研究[J]. 眼科, 2015, 24(3): 192-195.

SONG Si-yue, MA Ke, ZHAO Xiu-li. Neuroprotection of siRNA interference to the retinal ganglion cells in optic nerve crush model rat[J]. Ophthalmology in China, 2015, 24(3): 192-195.

[1]	尹鹏石砚王一玮王怀州辛晨曹凯万月王宁利. 微导管辅助的内路部分黏小管成形联合内路部分小梁切开术与内路全周小梁切开术的早期疗效对比[J]. 眼科, 2020, 29(4): 304-308.
[2]	刘朝唐耀冰杨延芳 . miR-130b通过调控PTEN在人视网膜母细胞瘤发挥促癌作用[J]. 眼科, 2017, 26(4): 276-281.
[3]	魏文斌曾司彦. 相干光断层扫描血管成像技术临床应用前景[J]. 眼科, 2015, 24(3): 145-148.
[4]	吴玲玲. 重视局部抗青光眼药物对眼表的影响[J]. 眼科, 2015, 24(3): 149-151.
[5]	陶远刘英. 不含苯扎氯铵的曲伏前列素滴眼液对患者干眼症状及泪膜质量的影响[J]. 眼科, 2015, 24(3): 153-155.
[6]	范媛媛阮余霞杨新. 视屏终端工作者泪液中基质金属蛋白酶-9、组织金属蛋白酶抑制剂-1活性研究[J]. 眼科, 2015, 24(3): 156-160.
[7]	贾力蕴赵博文桑景荭张妮红王宁利. 0.2%酒石酸溴莫尼定联合0.25%倍他洛尔的短期降眼压疗效及安全性研究 [J]. 眼科, 2015, 24(3): 160-163.
[8]	齐颖周跃华张晶张丽. VisxStarS4准分子激光机治疗近视眼实际切削深度与预计值差值的比较 [J]. 眼科, 2015, 24(3): 164-166.
[9]	要博辛志坤. 术中应用苏拉明与紫杉醇对兔小梁切除术后短期抗增生作用的对比研究 [J]. 眼科, 2015, 24(3): 167-171.
[10]	田蓓朱晓青. 视网膜色素变性393例临床特征分析 [J]. 眼科, 2015, 24(3): 177-181.
[11]	孙晓伟崔卉殷晓贝李根林. 原发性视网膜色素变性患者五种血清离子浓度的变化 [J]. 眼科, 2015, 24(3): 183-187.
[12]	李玉洁张海娟王海燕马科. 天麻钩藤饮抗大鼠视神经夹伤模型视网膜神经节细胞凋亡作用的实验研究[J]. 眼科, 2015, 24(3): 188-192.
[13]	禚伟王毅肖利华. 眼部Wegener肉芽肿17例临床分析[J]. 眼科, 2015, 24(3): 196-200.
[14]	柴歆孙婷郝琳娜肖媛媛李辽青李毅斌. 2型糖尿病患者糖尿病视网膜病变的家族聚集性研究 [J]. 眼科, 2015, 24(3): 202-205.
[15]	李丹孙挥宇毛菲菲王胜男柳月红许雪静刘彬彬董俞. 获得性免疫缺陷综合征合并巨细胞病毒性视网膜炎71例临床分析[J]. 眼科, 2015, 24(3): 206-209.

小干扰RNA保护大鼠视网膜神经节细胞的实验研究

Neuroprotection of siRNA interference to the retinal ganglion cells in optic nerve crush model rat

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价