摘要
目的观察单纯视神经损伤及视神经损伤联合晶状体损伤后视网膜神经节细胞及其轴突中锌指样转录因子4(KLF4)mRNA表达量的变化,探讨晶状体损伤促进视神经再生的相关机制。方法取54只健康成年Wistar大鼠随机分成对照组(6只)、单纯视神经损伤组(24只)、视神经损伤联合晶状体损伤组(24只)。分别于7d、14d、21d处死对照组大鼠各2只,单纯视神经损伤组和联合晶状体损伤组大鼠各8只。透射电镜观察损伤后轴突的显微结构变化,荧光定量聚合酶链反应检测不同分组不同时间点视网膜神经节细胞及其轴突上KLF4 mRNA的表达。结果损伤7d时,透射电镜下见单纯视神经损伤组和联合晶状体损伤组轴突较对照组明显肿胀,轴浆电子密度降低,髓鞘结构混乱,联合晶状体损伤组可见部分簇状排列的无髓鞘包裹的新生神经纤维,损伤14d和21d时单纯视神经损伤组和联合晶状体损伤组均少见髓鞘轮廓,见众多胶原纤维。损伤7d时,单纯视神经损伤组(2-△△Ct为0.561±0.045)和联合晶状体损伤组(0.091±0.026)KLF4 mRNA相对表达量较对照组(1.003±0.113)均有不同程度降低,联合晶状体损伤组降至最低,差异均有统计学意义(均为P<0.05);损伤14d时,单纯视神经损伤组(2-△△Ct为0.333±0.106)和联合晶状体损伤组(0.401±0.169)KLF4 mRNA相对表达量较对照组仍有不同程度降低,差异有统计学意义(均为P<0.05),但单纯视神经损伤组和联合晶状体损伤组间差异无统计学意义(P>0.05);损伤21d时,单纯视神经损伤组(2-△△Ct为1.100±0.100)和联合晶状体损伤组(1.166±0.115)KLF4 mRNA相对表达量较对照组轻度增高,差异均无统计学意义(均为P>0.05)。结论晶状体损伤促进视神经再生的机制可能与KLF4表达减少有关,这为基因转录水平靶向治疗视神经病变提供了新的思路。
Objective To observe the expression of Kruppel-like transcription factor 4 （KLF4） mRNA in rats with pure optic nerve injury and optic nerve injury combined with lens injury, and the relative mechanism of lens injury promoting optic nerve regeneration. Methods A total of fifty-four healthy Wistar rats were divided into three groups randomly ：normal group（6 rats） ,pure optic nerve injury group（24 rats） and optic nerve injury combined lens injury group（24 rats）. At 7 days,14 days and 21 days af- ter injury, two rats in normal control group, 8 rats in pure optic nerve injury group and 8 rats in optic nerve injury combined lens injury group were respectively ldlled. The microstructure of axon was observed by transmission electron microscope, and the expression of KLF4 mRNA in three groups at different time point was detected by fluorescent quantitative PCR. Results Compared with the normal group, EMS showed the obvious swelling of axon, axoplasm electron density becoming lower and chaos of myelin sheath structure, which were all visible in two experimental groups at 7 days after injury. EMS also showed some new nerve fibers without myelin sheath were partially clustered in combined injury group at 7 days. However, there were many collagen fibers and little myelin sheath in two experimental groups at 14 days and 21 days. Compared with the normal group（ 1. 003±0. 113） ,the relative expression of KLF4 mRNA in pure injury group（0.551±0.045） and combined injury group（0.091±0.026） were all decreased at different degree, which reduced to the minimum in combined injury group at 7 days, and there was statistically signiflcant（P 〈0.05 ） ;compared with the normal group,the relative expression of KLF4 mRNA in pure injury group （ 0. 333 ± 0. 106 ） and combined injury group（0.401±0. 159） also decreased at some degree at 14 days,and there was statistically significant（P 〈 0.05 ）, but there was no statistical significance between the pure injury group and combined injury group（P〉 0. 05 ） ; compared with the normal group,the expression of KLF4 mRNA in pure injury group（ 1. 100 ± 0. 100） and combined injury group（ 1. 155 ± 0. 115 ） slightly increased,there was no statistical significance among normal group,pure injury group and combined injury group （ all P 〉 0.05 ） at 21 days. Conclusion Lens injury promoting optic nerve regeneration mechanism may relate to the reducing KLF4 expression, which provides a new approach to targeted therapy neuropathy in gene transcription level.
出处
《眼科新进展》
CAS
北大核心
2013年第5期431-434,共4页
Recent Advances in Ophthalmology
基金
河南省重点科技攻关项目(编号:072102310116)~~
关键词
锌指样转录因子4
晶状体损伤
视神经损伤
视神经再生
Kruppel-like transcription factor 4
lens injury
optic nerve injury
optic nerve regeneration
