The Study Of MicroRNA Expression In The Aqueous Humor Of Highly Myopic Patients And Its Mechanism

BackgroundHigh myopia has been increasing in recent years.At present,the prevalence of high myopia among children and adolescents in China has been 3.1%.It is estimated that the prevalence of high myopia worldwide will be 10% by 2050.With the optical axis elongation of patients with high myopia,serious complications such as macular hemorrhage and posterior scleral staphyloma may occur,which results in irreversible damage and loss of vision.At present,the most popular theory about the pathogenesis of myopia is the theory of scleral remodeling.In the study of animal models of myopia,it was found that the most important pathological change during myopia was scleral extracellular matrix remodeling(ECM).Scleral ECM remodeling during myopia is mainly due to decreased synthesis and increased degradation of ECM,especially collagen I.However,what causes myopia-related scleral remodeling remains unclear.One hypothesis is that visual signals are converted into signaling molecules in the retina,transmitted to the choroid,and then to the sclera,leading to Scleral ECM remodeling.micro RNA(mi RNA)are non-coding RNA that silence the expression of target genes at the post-transcriptional level and are widely involved in pathological processes such as cancer and inflammation.More and more studies have shown that mi R-142-3p is an important molecule regulating ECM and can inhibit the expression of ECM in various tissues.However,it is unclear whether mi R-142-3p can regulate the synthesis of scleral ECM.TGF-β1 plays a key role in regulating ECM.In the sclera of myopic animal models,the content of TGF-β1 is reduced,which leads to the reduction of collagen during myopia.However,no study has reported whether mi RNA are involved in the regulation of TGF-β1 in the process of myopia.This study will research the pathogenesis of high myopia from the aspect of mi RNA,and find new targets for the prevention and treatment of high myopia.Method1.The myopic patients were divided into high myopia group(diopter≥-6.00 D,axial length≥26mm)and control group(diopter-0.50D～-6.00 D,axial length 24～26mm).High-throughput sequencing was used to detect the expression profiles of mi RNA in the aqueous humor of these two groups.Then we screened the differentially expressed mi RNA in high myopia patients,and predicted the target genes of these mi RNA.Bioinformatics analysis of these target genes was conducted.2.Quantitative PCR(q PCR)was used to detect the expression of hsa-mi R-142-3p in the aqueous humor of patients with high myopia,and then we analyze the relationship between the expression of hsa-mi R-142-3p and ocular axial length,diopter,retinal thickness and choroidal thickness.3.We Overexpressed and inhibited hsa-mi R-142-3p in human fetal scleral fibroblasts(HFSFs)to explore its effect on the synthesis of ECM.3.We Overexpressed and inhibited hsa-mi R-142-3p in HFSFs to explore its regulation on TGF-β1,and used luciferase reporter assay to confirm the binding site of hsa-mi R-142-3p and TGF-β1.Finally,TGF-β1 was added to human scleral fibroblasts transfected with mi R-142-3p to explore the effect of TGF-β1on the inhibition of extracellular matrix synthesis by mi R-142-3p.Result1.High-throughput sequencing detected the mi RNA expression profiles of the high myopia group and control group.37 differentially expressed mi RNAs between high myopia group and control group were screened out.Compared with the control group,there were 19 mi RNA upregulated and 18 mi RNA down-regulated in high myopia group.2.Hsa-mi R-142-3p in the aqueous humor of high myopia patients was higher than that of control group.The expression of hsa-mi R-142-3p in the aqueous humor of myopic patients was positively correlated with the ocular axial length and diopter,but has no correlation with retinal thickness and choroidal thickness.3.Hsa-mi R-142-3p overexpression reduced collagen I in the HFSFs,while the hsa-mi R-142-3p inhibitor increased collagen I.4.Hsa-mi R-142-3p targeted TGF-β1.TGF-β1 can reverse the inhibition of mi R-142-3p on collagen I in HFSFs.ConclusionThe expression of hsa-mi R-142-3p increases during high myopia,and hsa-mi R-142-3p suppresses collagen I in scleral fibroblasts by silencing the target gene TGF-β1,resulting in scleral ECM remodeling and sclera thinning,which promotes optical axis elongation and development of high myopia.