衰老相关长链非编码RNA PINT通过表观遗传学修饰影响年龄相关性白内障的发生发展

Senescence-associated LncRNA PINT influences the development of age-related cataracts through epigenetic modifications

目的探讨衰老相关长链非编码RNA(LncRNA)PINT对年龄相关性白内障(ARC)发生发展的影响。方法通过收集不同类型ARC及透明晶状体的上皮细胞,测定衰老相关LncRNA的表达;通过对比ARC组和对照组,筛选衰老相关的差异表达LncRNA PINT。采用实时荧光定量PCR(qRT-PCR)检测ARC组与对照组晶状体上皮细胞中LncRNA PINT的表达水平。使用微阵列测定LINC-PINT的潜在靶基因,以及使用染色质免疫共沉淀(ChIP)验证LncRNA PINT靶基因候选物的状态。建立氧化损伤/衰老模型、过表达/敲低模型,检测LncRNA PINT及相关靶基因的表达水平、细胞抗氧化损伤能力、细胞增殖凋亡情况。结果ARC组中LncRNA PINT的表达量高于对照组,差异有统计学意义(P<0.05);氧化损伤/衰老模型中LncRNA PINT的表达量高于对照组,差异有统计学意义(P<0.05)。敲低模型中LncRNA PINT相关靶基因表达水平显著升高,晶状体上皮细胞抗氧化能力增强(P<0.05);过表达模型中晶状体上皮细胞增殖减少,凋亡增加(P<0.05)。结论衰老相关LncRNA的表达改变可能通过表观遗传学修饰影响机体的衰老和衰老性疾病的形成,与ARC的发生发展相关。

Objective To investigate the effect of senescence-associated long non-coding RNA(LncRNA)PINT on the development of age-related cataract(ARC).Methods The expression of aging-related LncRNAs was determined by collecting epithelial cells from different types of ARC and clear lens,and the differentiated expression aging-related LncRNA PINT was screened by comparing the ARC group with the control group.Quantitative Real-time PCR(qRT-PCR)was performed to detect the expression levels of LncRNA PINT in the lens epithelial cells of the ARC group and the control group.Microarray assays was used to identify potential LncRNA PINT target genes,and chromatin immunoprecipitation assay(ChIP)was used to verify the status of LncRNA PINT target gene candidates.Oxidative damage/aging model and overexpression/knockdown model were established to detect the expression levels of LncRNA PINT and related target genes,cellular antioxidant damage capacity,cell proliferation and apoptosis.Results The expression level of LncRNA PINT in ARC group was significantly higher than that in control group(P<0.05).The expression level of LncRNA PINT in oxidative damage/aging model was significantly higher than that in control group(P<0.05).In knockdown model,the expression level of LncRNA PINT related target genes was significantly increased,and the antioxidant capacity of lens epithelial cells was enhanced(P<0.05).In the overexpression model,lens epithelial cells showed decreased proliferation and increased apoptosis(P<0.05).Conclusion Changes in expression of senescence-associated LncRNAs may affect aging and senile disease formation in the organism through epigenetic modifications,which are associated with the development of ARC.