[1]李晶,曾卫娟.LncRNA HAGLR靶向miR-625-5p对脂多糖诱导的视网膜色素上皮细胞凋亡和炎症因子表达的影响[J].眼科新进展,2024,44(3):178-182.[doi:10.13389/j.cnki.rao.2024.0035]
　LI Jing,ZENG Weijuan.Effect of long-chain non-coding ribonucleic acid HAGLR targeting micro ribonucleic acid-625-5p on lipopolysaccharide-induced apoptosis and inflammatory factor expression in retinal pigment epithelial cells[J].Recent Advances in Ophthalmology,2024,44(3):178-182.[doi:10.13389/j.cnki.rao.2024.0035]
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LncRNA HAGLR靶向miR-625-5p对脂多糖诱导的视网膜色素上皮细胞凋亡和炎症因子表达的影响/HTML
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《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
44卷
期数:
2024年3期
页码:
178-182
栏目:
实验研究
出版日期:
2024-03-05

文章信息/Info

Title:
Effect of long-chain non-coding ribonucleic acid HAGLR targeting micro ribonucleic acid-625-5p on lipopolysaccharide-induced apoptosis and inflammatory factor expression in retinal pigment epithelial cells
作者:
李晶; 曾卫娟
457000　河南省濮阳市，濮阳医学高等专科学校(李晶);430062　湖北省武汉市，武汉大学中南医院眼科（曾卫娟）
Author(s):
LI Jing1; ZENG Weijuan2
1.Puyang Medical College,Puyang 457000,Henan Province,China
2.Department of Ophthalmology,Zhongnan Hospital of Wuhan University,Wuhan 430062,Hubei Province,China
关键词:
长链非编码RNA HAGLR; 微小RNA-625-5p; 脂多糖; 视网膜色素上皮细胞; 细胞凋亡; 炎症
Keywords:
long-chain non-coding ribonucleic acid HAGLR;  micro ribonucleic acid-625-5p;  lipopolysaccharide;  retinal pigment epithelial cells;  apoptosis;  inflammation
分类号:
R774.1
DOI:
10.13389/j.cnki.rao.2024.0035
文献标志码:
A
摘要:
目的　探讨长链非编码RNA HAGLR（LncRNA HAGLR）是否可通过靶向调控微小RNA-625-5p（miR-625-5p）表达而影响脂多糖（LPS）诱导的视网膜色素上皮（RPE）细胞凋亡、炎症因子表达，为揭示视网膜病变机制奠定实验基础。
方法　将LPS诱导人视网膜色素上皮细胞（ARPE-19）设为LPS组，正常培养的ARPE-19细胞设为Con组。实时荧光定量聚合酶链反应（qRT-PCR）检测LncRNA HAGLR、miR-625-5p表达水平。根据转染物不同分为LPS+sh-NC组、LPS+sh-HAGLR组、LPS+miR-NC组、LPS+miR-625-5p组、LPS+sh-HAGLR+anti-miR-NC组、LPS+sh-HAGLR+anti-miR-625-5p组。采用流式细胞术检测细胞凋亡率；ELISA法检测白细胞介素-6（IL-6）、白细胞介素-1β（IL-1β）水平；验证LncRNA HAGLR、miR-625-5p靶向关系；Western blot检测活化的半胱氨酸天冬氨酸特异性蛋白酶3（cleaved-Caspase 3)、cleaved-Caspase 9蛋白水平。
结果　与Con组比较，LPS组LncRNA HAGLR表达水平升高，miR-625-5p表达水平降低（均为P<0.05），细胞凋亡率，cleaved-Caspase 3、cleaved-Caspase 9蛋白水平，IL-6、IL-1β水平均升高（均为P<0.05）。与LPS+sh-NC组比较，LPS+sh-HAGLR组细胞凋亡率，cleaved-Caspase 3、cleaved-Caspase 9蛋白水平，IL-6、IL-1β水平均降低（均为P<0.05）；LncRNA HAGLR可负向调控miR-625-5p表达水平（P<0.05）。与LPS+miR-NC组比较，LPS+miR-625-5p组miR-625-5p表达水平升高，细胞凋亡率，cleaved-Caspase 3、cleaved-Caspase 9蛋白水平，IL-6、IL-1β水平均降低（均为P<0.05）。与LPS+sh-HAGLR+anti-miR-NC组比较，LPS+sh-HAGLR+anti-miR-625-5p组miR-625-5p表达水平降低，细胞凋亡率，cleaved-Caspase 3、cleaved-Caspase 9蛋白水平，IL-6、IL-1β水平均升高（均为P<0.05）。
结论　干扰LncRNA HAGLR表达可通过靶向调控miR-625-5p表达抑制细胞凋亡、炎症因子表达，以减轻LPS诱导的ARPE-19细胞损伤。
Abstract:
Objective　To investigate whether long-chain non-coding ribonucleic acid HAGLR (LncRNA HAGLR) can affect lipopolysaccharide (LPS)-induced apoptosis and expression of inflammatory factors of retinal pigment epithelium (RPE) cells by targeted regulation of the expression of micro ribonucleic acid-625-5p (miR-625-5p), so as to lay an experimental foundation for revealing the mechanism of retinopathy.
Methods　LPS-induced human retinal pigment epithelial (ARPE-19) cells were set as the LPS group, and normally cultured ARPE-19 cells were assigned to the Con group. Quantitative real-time polymerase chain reaction was used to detect the expression levels of LncRNA HAGLR and miR-625-5p. Based on different transfection reagents, the cells were divided into the LPS+sh-NC group, LPS+sh-HAGLR group, LPS+miR-NC group, LPS+miR-625-5p group, LPS+sh-HAGLR+anti-miR-NC group, and LPS+sh-HAGLR+anti-miR-625-5p group. Flow cytometry was used to detect apoptosis rate; enzyme-linked immunosorbent assay was used to detect the levels of interleukin-6 (IL-6) and interleukin-1β (IL-1β); the targeting relationship between LncRNA HAGLR and miR-625-5p was verified; Western blot was used to detect the protein levels of activated cysteinyl aspartate specific proteinase 3 and 9 (cleaved-Caspase 3 and cleaved-Caspase 9).
Results　Compared with the Con group, the LPS group showed an increase in the expression level of LncRNA HAGLR and a decrease in the expression level of miR-625-5p (both P<0.05), and there were increases in apoptosis rate, protein levels of cleaved-Caspase 3 and cleaved-Caspase 9, and levels of IL-6 and IL-1β (all P<0.05). Compared with the LPS+sh-NC group, the LPS+sh-HAGLR group showed decreases in apoptosis rate, protein levels of cleaved-Caspase 3 and cleaved-Caspase 9, and levels of IL-6 and IL-1β (all P<0.05); LncRNA HAGLR could negatively regulate the expression level of miR-625-5p (P<0.05). Compared with the LPS+miR-NC group, the LPS+miR-625-5p group showed an increase in the expression level of miR-625-5p and decreases in apoptosis rate, protein levels of cleaved-Caspase 3 and cleaved-Caspase 9, and levels of IL-6 and IL-1β (all P<0.05). Compared with the LPS+sh-HAGLR+anti-miR-NC group, the LPS+sh-HAGLR+anti-miR-625-5p group showed a decrease in the expression level of miR-625-5p and increases in apoptosis rate, protein levels of cleaved-Caspase 3 and cleaved-Caspase 9, and levels of IL-6 and IL-1β (all P<0.05).
Conclusion　Interference on LncRNA HAGLR expression can realize the targeted regulation of miR-625-5p expression to inhibit the apoptosis and inflammatory factor expression, reducing LPS-induced injury of ARPE-19 cells.

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备注/Memo

备注/Memo:
国家卫生健康委“十三五”规划全国重点课题(编号:YYWS2330)
更新日期/Last Update: 2024-03-05