LncRNA-MIAT靶向miR-150-5p对病理性血管化角膜及VEGF、MMP-9的影响

Effect of LncRNA-MIAT targeting miR-150-5p on pathological vascularized cornea,vascular endothelial growth factor,and matrix metalloproteinase-9

目的探讨长链非编码核糖核酸心肌梗死相关转录本(LncRNA-MIAT)靶向miR-150-5p对病理性血管化角膜血管内皮生长因子(VEGF)及基质金属蛋白酶-9(MMP-9)的影响。方法100只大鼠随机分为对照组、模型组、MIAT-NC组、MIAT组和sh-MIAT组,除对照组外,其余大鼠以碱烧伤法建立角膜血管化模型。实时荧光定量PCR(RT-PCR)检测大鼠角膜组织中LncRNA-MIAT和miR-150-5p相对表达情况;双荧光素酶实验验证LncRNA-MIAT和miR-150-5p的靶向关系;ELISA检测大鼠角膜组织中肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-6、IL-8水平;HE染色观察大鼠角膜组织中血管病理变化;Western blot检测大鼠角膜组织中VEGF、MMP-9相对表达情况。结果与对照组比较,模型组大鼠角膜组织中LncRNA-MIAT相对表达水平,TNF-α、IL-6、IL-8水平,VEGF和MMP-9蛋白相对表达量均升高,miR-150-5p相对表达水平降低(均为P<0.05);与模型组和MIAT-NC组比较,MIAT组大鼠角膜组织中LncRNA-MIAT相对表达水平,TNF-α、IL-6、IL-8水平,VEGF和MMP-9蛋白相对表达量均升高,miR-150-5p相对表达水平降低(均为P<0.05),sh-MIAT组上述指标与MIAT组呈相反趋势(均为P<0.05)。双荧光素酶实验结果显示,LncRNA-MIAT可靶向调控miR-150-5p表达。结论LncRNA-MIAT可促进VEGF、MMP-9的表达,加速角膜血管化的发生,其机制可能与抑制miR-150-5p的表达有关。

Objective To investigate the effect of long non-coding ribonucleic acid myocardial infarction-associated transcript(LncRNA-MIAT)targeting microribonucleic acid-150-5p(miR-150-5p)on vascular endothelial growth factor(VEGF)in pathological vascularized cornea and matrix metalloproteinase-9(MMP-9).Methods A total of 100 rats were randomly divided into the control group,model group,MIAT-NC group,MIAT group,and sh-MIAT group.Except for the control group,rats in other groups were used to establish a corneal vascularization model by the alkali burn method.The real-time polymerase chain reaction(RT-PCR)was used to detect the relative expression of LncRNA-MIAT and miR-150-5p in the cornea of rats,the dual-luciferase assay was used to verify the targeting relationship between LncRNA-MIAT and miR-150-5p,the ELISA was used to detect corneal tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),and interleukin-8(IL-8)levels,the hematoxylin and eosin(HE)staining was used to observe the pathological changes of corneal blood vessels,and the Western blot assay was used to detect the relative expression of VEGF and MMP-9 in corneal tissues.Results Compared with the control group,the relative expression of LncRNA-MIAT,the levels of TNF-α,IL-6 and IL-8,and the protein expression of VEGF and MMP-9 in the model group increased,while the relative expression of miR-150-5p decreased(all P<0.05).Compared with the model group and the MIAT-NC group,the relative expression of LncRNA-MIAT,the levels of TNF-α,IL-6 and IL-8,and the protein expression of VEGF and MMP-9 in the MIAT group increased,while the relative expression of miR-150-5p decreased(all P<0.05).The above indicators in the sh-MIAT group showed an opposite trend to the MIAT group(all P<0.05).The dual-luciferase assay results showed that LncRNA-MIAT could target the expression of miR-150-5p.Conclusion LncRNA-MIAT can promote the expression of VEGF and MMP-9 to accelerate the occurrence of corneal vascularization.The mechanism may be related to the inhibited expression of miR-150-5p.