| Objective:To explore the possible mechanism of lens-retina-optic nerve interaction in promoting optic nerve regeneration after lens injuryMethods:SD rats were dilated by topicamide eye drops,and the anterior capsular membrane of the lens was punctured with a 30 G needle to make the rat model of lens injury.After the optic nerve was exposed,the optic nerve injury model was made by clamping.Experimental animals were divided into Control group(Control group),optic nerve injury combined with contralateral eye lens injury group(ON-L opposite group),optic nerve injury group(ON group),with the side joint lens damage nerve injury group(ON-L group).The up-regulated expression of MMP-8 and MMP-12 was used as markers of optic nerve regeneration.The gene and protein expressions of MMP-8 and MMP-12 at 7 days,14 days,21 days and 28 days were detected by RT-q PCR and ELISA.According to the results of presequence experiment,the animal models of optic nerve injury combined with lens injury group(ON-L),optic nerve injury group(ON)and lens injury group(L)were reconstructed.Total RNA was extracted from retinal tissue on the 7th day,and reference transcriptome sequencing was performed by high-throughput sequencing technology,gene difference analysis,GO and KEGG enrichment analysis,etc.Based on transcriptome sequencing data,follow-up verification experiments were preliminarily conducted to speculate the possible mechanism.Results:1.RT-qPCR: Compared with ON group,the opposite group of ON-L could not increase the expression of MMP-8 and MMP-12 at all time points ON days 7,14,21 and 28.ON-L group showed significantly high expression at all time points.2.ELISA detection: Protein quantitative detection was consistent with the results of qRT-PCT.Compared with ON group,the expression of MMP-8 and MMP-12 in ON-L opposite group was not up-regulated at all time points ON day 7,day 14,day 21 and day 28.ON-L group showed significantly high expression at all time points.3.Quality control analysis: The clean read statistical analysis formed by filtering the original sequencing data showed that the average base sequencing error rate was less than 0.01%,Q20 was more than 95.90%,Q30 was more than90% except ON-L2 group(89.9%),and GC content of each group was between47 and 54%.4.Analysis of gene differential expression: Because of the differences between the groups of genes(log2Fold Change | > 0 & padj < 0.05),the number is too big,bad for screening,using | log2 Fold Change | > 1 & padj < 0.01 &basemean > 100 as more stringent eligibility,screening of gene has significant differences.5.GO and KEGG enrichment analysis: Based on neuronutrition,protection,cell regeneration and differentiation,GO and KEGG were screened to obtain differential genes related to this study.Conclusion:1.Lens injury induces long-term regeneration of the optic nerve after injury through local effects.2.The MAPK pathway may be the key pathway for the protection of retinal ganglion cells during the long-term regeneration of optic nerve induced by lens injury.3.Gadd45 g and Hspb1 may be the key factors for the action of MAPK pathway on retina.4.After lens injury induced by optic nerve regeneration in the process,may activate the axoplasmic transport function of the retina tissue related Obsl1 and Slit3 factor... |
