在晶状体损伤诱导的视神经长时程再生过程中MMP-12的表达规律

Expression pattern of MMP-12 in long-term optic nerve regeneration induced by lens injury

摘要:

目的：探讨晶状体损伤诱导SD大鼠视神经钳压损伤后轴突长时程再生过程中MMP-12的表达变化。方法：建立SD大鼠视神经损伤模型和晶状体损伤模型，将实验动物24只分为对照组(仅开放眼眶暴露视神经)、晶状体损伤组、视神经损伤组、晶状体损伤联合视神经损伤组，每组各6只大鼠。采用有参转录组测序分析损伤视神经区域差异基因表达变化，筛选相关高表达差异基因，通过实时荧光定量PCR(qRT-PCR)和酶联免疫吸附实验(ELISA)定量分析损伤区MMP-12的表达量变化。结果：转录组测序主成因分析表明，晶状体损伤联合视神经损伤是基因表达变化中的主要成因。基因表达差异分析显示，晶状体损伤联合视神经损伤组中，存在MMP-12基因表达上调。在建模成功后14、21d时，晶状体损伤联合视神经损伤组的MMP-12 mRNA表达量与对照组、视神经损伤组和晶状体损伤组比较上调(P<0.05)； 在7、28d时，各组间表达无差异。在建模成功后7、14、21d时，晶状体损伤联合视神经损伤组的MMP-12蛋白表达量与对照组和视神经损伤组比较上调(P<0.05)； 21d时，晶状体损伤联合视神经损伤组与视神经损伤组比较上调(P<0.05)。28d时，各组间表达无差异。结论：MMP-12表达上调可能参与晶状体损伤后诱导视神经长时程再生过程。

Abstract:

AIM: To investigate the expression changes of MMP-12 during the long-term axon regeneration induced by the lens injury after the optic nerve clamp trauma in sprague-dawley(SD)rats.METHODS: The optic nerve injury model and lens injury model of SD rats were established, and the 24 experimental animals were divided into control group; lens injury group; optic nerve injury group; lens injury combined with optic nerve injury group, with 6 rats in each group. Reference transcriptome sequencing was used to analyze the expression changes of differentially expressed genes in the injured optic nerve region, and relevant differentially expressed genes with high expression were screened. Quantitative real-time polymerase chain reaction(qRT-PCR)and enzyme-linked immunosorbent assay(ELISA)were used to quantify the expression changes of matrix metalloproteinase-12(MMP-12)in the injured optic nerve region.RESULTS: The Principal Component Analysis of transcriptome sequencing indicated that lens injury combined with optic nerve injury was the principal component of gene expression change. Analysis of gene expression differences showed that the expression of MMP-12 gene was up-regulated in the lens injury combined with optic nerve injury group. The mRNA expression level of MMP-12 in the lens injury combined optic nerve injury group was up-regulated compared with the control group, the optic nerve injury group and the lens injury group at 14d and 21d after successful modeling(P<0.05). At 7, 28d, there was no difference in expression among all groups. The protein expression level of MMP-12 in the lens injury combined with optic nerve injury group was up-regulated compared with the control group and optic nerve injury group at 7, 14 and 21d after successful modeling(P<0.05), and it was up-regulated in the lens injury group combined with optic nerve injury group compared with optic nerve injury group at 21d(P<0.05). At 28d, there was no difference in expression among all groups.CONCLUSION: The up-regulated expression of MMP-12 may be involved in the long-term regeneration of the optic nerve after lens injury.