目的研究高糖培养条件下人视网膜血管内皮细胞(HRVECs)的环状RNA(circRNA)表达变化。
方法将HRVECs分为正常对照组、高渗对照组和高糖组，分别在5.5 mmol/L葡萄糖培养基、19.5 mmol/L甘露醇+5.5 mmol/L葡萄糖培养基和25 mmol/L葡萄糖培养基中培养24 h。使用circRNA芯片对高糖组和正常对照组HRVECs进行circRNA表达谱检测，筛选出差异表达circRNA分子；采用实时荧光定量PCR验证差异表达circRNA分子在正常对照组、高渗对照组和高糖组细胞中的表达变化，并采用Circular RNA Interactome数据库预测差异表达circRNA可能作用的微小RNA(miRNA)靶点。
结果在高糖培养的HRVECs中共筛选出448个差异表达circRNA(差异倍数≥1.5或≤0.67且 P<0.05)，其中182个circRNA上调，266个circRNA下调。表达上调排序前3的差异circRNA分别为hsa_circ_0002938、hsa_circ_0008036和hsa_circ_0001946，表达下调排序前3的差异circRNA分别为hsa_circ_0035277、hsa_circ_0008344和hsa_circ_0001874。实时荧光定量PCR结果显示，与正常对照组和高渗对照组相比，高糖组中hsa_circ_0002938、hsa_circ_0008036和hsa_circ_0001946相对表达量显著升高，hsa_circ_0035277、hsa_circ_0008344和hsa_circ_0001874相对表达量显著下调，差异均有统计学意义(均 P<0.05)；正常对照组与高渗对照组各差异circRNA表达比较，差异均无统计学意义(均 P>0.05)。
结论在高糖条件下培养的HRVECs中circRNA呈差异表达，差异表达circRNA可能参与糖尿病视网膜病变发病机制。
ObjectiveTo investigate the differential expression profile of circular RNA (circRNA) in high glucose-cultured human retinal vascular endothelial cells (HRVECs).
MethodsHRVECs were divided into high glucose group, normal control group and hypertonic control group, and were cultured in 25 mmol/L glucose medium, 5.5 mmol/L glucose medium and 19.5 mmol/L mannitol+ 5.5 mmol/L glucose medium for 24 hours accordingly.The differentially expressed circRNA molecules between high glucose group and normal control group were screened by circRNA microarray analysis.The expression of the most significant differentially expressed circRNAs in different groups was verified by real-time quantitative PCR.The possible microRNA (miRNA) targets were analyzed through the Circular RNA Interactome database.
ResultsIt was found that 448 circRNAs were differentially expressed (FC≥1.5 or FC≤0.67, P<0.05) in high glucose-cultured HRVECs, among which 182 were up-regulated and 266 were down-regulated.The top 3 significantly up-regulated circRNAs were hsa_circ_0002938, hsa_circ_0008036, and hsa_circ_0001946, and the top 3 significantly down-regulated circRNAs were hsa_circ_0035277, hsa_circ_0008344, and hsa_circ_0001874.Compared with normal control group and hypertonic control group, the relative expressions of top 3 up-regulated circRNAs were significantly enhanced and the relative expressions of top 3 down-regulated circRNAs were significantly reduced in high glucose group, showing statistically significant differences (all at P<0.05).No significant difference was found in the differentially expressed circRNAs between normal control group and hypertonic control group (all at P>0.05).
ConclusionsCircRNAs are differentially expressed in high glucose-cultured HRVECs, and the differentially expressed circRNAs may be involved in the regulatory mechanism of diabetic retinopathy.
贾杨雪,王志玲,魏莹莹,等. 高糖诱导的人视网膜血管内皮细胞中环状RNA差异表达分析[J]. 中华实验眼科杂志,2022,40(07)：617-622.
DOI:10.3760/cma.j.cn115989-20200602-00390版权归中华医学会所有。
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贾杨雪：实验实施、数据整理及分析、论文撰写；王志玲、魏莹莹、朱恺：数据整理和分析；顾永昊：实验设计、论文修改及定稿
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