Effect of lncRNA KCNQ1OT1 on oxygen glucose deprivation/reoxygenation-induced microglia injury through regulating the miR-145-5p/ROCK1 axis

LI Lin; LI Xiang-nan; YANG Song-tao; ZHANG Yun-he; CHEN Tong; FU Ai-jun

doi:10.11659/jjssx.06E024007

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Effect of lncRNA KCNQ1OT1 on oxygen glucose deprivation/reoxygenation-induced microglia injury through regulating the miR-145-5p/ROCK1 axis

Basic Research | 更新时间：2025-06-17

- Effect of lncRNA KCNQ1OT1 on oxygen glucose deprivation/reoxygenation-induced microglia injury through regulating the miR-145-5p/ROCK1 axis
- JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY Vol. 34, Issue 6, Pages: 478-483(2025)
- 作者机构：
  
  华北理工大学附属医院神经外科，河北唐山 063000
- 作者简介：
- 基金信息：
- DOI：10.11659/jjssx.06E024007
  CLC： R743.3
- Received：03 June 2024，
  
  Published：25 June 2025
- 稿件说明：
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李林,李向男,杨松涛等.lncRNA KCNQ1OT1调节miR-145-5p/ROCK1轴对氧糖剥夺/复氧诱导的小胶质细胞损伤的影响[J].局解手术学杂志,2025,34(06):478-483.

LI Lin,LI Xiang-nan,YANG Song-tao,et al.Effect of lncRNA KCNQ1OT1 on oxygen glucose deprivation/reoxygenation-induced microglia injury through regulating the miR-145-5p/ROCK1 axis[J].JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY,2025,34(06):478-483.
李林,李向男,杨松涛等.lncRNA KCNQ1OT1调节miR-145-5p/ROCK1轴对氧糖剥夺/复氧诱导的小胶质细胞损伤的影响[J].局解手术学杂志,2025,34(06):478-483. DOI： 10.11659/jjssx.06E024007.

LI Lin,LI Xiang-nan,YANG Song-tao,et al.Effect of lncRNA KCNQ1OT1 on oxygen glucose deprivation/reoxygenation-induced microglia injury through regulating the miR-145-5p/ROCK1 axis[J].JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY,2025,34(06):478-483. DOI： 10.11659/jjssx.06E024007.

摘要

目的

探讨长链非编码RNA（lncRNA）钾电压门控通道亚家族Q成员1反向转录物1（KCNQ1OT1）调节miR-145-5p/Rho相关卷曲螺旋形成蛋白激酶1（ROCK1）轴对氧糖剥夺/复氧（OGD/R）诱导的小胶质细胞损伤的影响。

方法

将小胶质细胞N9分为对照组（正常培养）、OGD/R组（OGD/R诱导损伤）、sh-NC组（OGD/R诱导损伤后转染sh-NC）、sh-KCNQ1OT1组（OGD/R诱导损伤后转染sh-KCNQ1OT1）、sh-KCNQ1OT1+inhibitor NC组（OGD/R诱导损伤后共转染sh-KCNQ1OT1和inhibitor NC）、sh-KCNQ1OT1+miR-145-5p inhibitor组（OGD/R诱导损伤后共转染sh-KCNQ1OT1和miR-145-5p inhibitor）。RT-qPCR检测细胞中lncRNA KCNQ1OT1、miR-145-5p、ROCK1 mRNA的表达；Western blot分析ROCK1蛋白表达；CCK-8测定细胞增殖能力；流式细胞术分析细胞凋亡情况；ELISA检测细胞中白细胞介素-1β（IL-1β）、肿瘤坏死因子-α（TNF-α）、白细胞介素-6（IL-6）水平；通过生物信息学网站预测miR-145-5p与lncRNA KCNQ1OT1、ROCK1的作用位点，并使用双荧光素酶报告基因实验验证其靶向关系。

结果

与对照组比较，OGD/R组细胞中lncRNA KCNQ1OT1表达、ROCK1 mRNA和蛋白表达、细胞凋亡率及IL-6、TNF-α、IL-1β水平均升高（

0.05），miR-145-5p表达、细胞存活率均降低（

0.05）；与OGD/R组、sh-NC组比较，sh-KCNQ1OT1组细胞中lncRNA KCNQ1OT1表达、ROCK1 mRNA和蛋白表达、细胞凋亡率及IL-6、TNF-α、IL-1β水平均降低，miR-145-5p表达、细胞存活率均升高，差异有统计学意义（

0.05）；与sh-KCNQ1OT1+inhibitor NC组比较，sh-KCNQ1OT1+miR-145-5p inhibitor组细胞中ROCK1 mRNA和蛋白表达、细胞凋亡率及IL-6、TNF-α、IL-1β水平均增加，miR-145-5p表达、细胞存活率均降低，差异有统计学意义（

0.05）。生物信息学网站预测显示，miR-145-5p与lncRNA KCNQ1OT1、ROCK1均存在靶向作

用位点；双荧光素酶报告基因实验表明miR-145-5p与lncRNA KCNQ1OT1和ROCK1存在靶向关系（

0.05）。

结论

沉默lncRNA KCNQ1OT1可通过上调miR-145-5p表达，靶向下调ROCK1表达，减轻OGD/R诱导的小胶质细胞损伤。

Abstract

Objective

To investigate the effect of long non-coding RNA（lncRNA） potassium voltage-gated channel subfamily Q member 1 overlapping transcript 1 （KCNQ1OT1） on oxidative glucose deprivation/reoxygenation （OGD/R）-induced microglia injury through regulating the miR-145-5p/ Rho-associated coiled-coil forming protein kinase 1 （ROCK1） axis.

Methods

Microglia N9 were divided into the control group （normal culture），the OGD/R group （OGD/R-induced injury），the sh-NC group （transfected with sh-NC after OGD/R-induced injury），the sh-KCNQ1OT1 group （transfected with sh-KCNQ1OT1 after OGD/R-induced injury），the sh-KCNQ1OT1+inhibitor NC group （co-transfected with sh-KCNQ1OT1 and inhibitor NC after OGD/R-induced injury），and the sh-KCNQ1OT1+miR-145-5p inhibitor group （co-transfected with sh-KCNQ1OT1 and miR-145-5p inhibitor after OGD/R-induced injury）. RT-qPCR was applied to detect the expression of lncRNA KCNQ1OT1， miR-145-5p， and ROCK1 mRNA in cells. The expression of ROCK1 protein was detected by Western blot. CCK-8 was applied to detect the cell proliferation. Flow cytometry was applied to detect cell apoptosis. ELISA was applied to detect the levels of interleukin-1β（IL-1β），tumor necrosis factor-α（TNF-α）， and interleukin-6（IL-6） in cells. The action sites of miR-145-5p with lncRNA KCNQ1OT1 and ROCK1 were predicted by bioinformatics website， and their targeting relationships were verified by dual-luciferase reporter assay.

Results

Compared with the control group， the expression of lncRNA KCNQ1OT1， the expression of ROCK1 mRNA and protein， the apoptosis rate， and the levels of IL-6， TNF-α， and IL-1β in cells were all increased， and the level of miR-145-5p and cell survival rate were all dec

reased in the OGD/R group（

0.05）. Compared with the OGD/R group and the sh-NC group， the expression of lncRNA KCNQ1OT1， the expression of ROCK1 mRNA and protein， the apoptosis rate，and the levels of IL-6，TNF-α，and IL-1β in cells were all decreased，and the level of miR-145-5p and cell survival rate were all increased in the sh-KCNQ1OT1 group，with significant differences （

0.05）. Compared with the sh-KCNQ1OT1+inhibitor NC group， the expression of ROCK1 mRNA and protein， the apoptosis rate，and the levels of IL-6， TNF-α， and IL-1β in cells were all increased， and the expression of miR-145-5p and cell survival rate were all decreased in the sh-KCNQ1OT1+miR-145-5p inhibitor group， with significant differences （

0.05）. Bioinformatics website showed that miR-145-5p had targeted action sites with lncRNA KCNQ1OT1 and ROCK1， and dual-luciferase reporter assay confirmed that miR-145-5p had targeting relationships with lncRNA KCNQ1OT1 and ROCK1 （

0.05）.

Conclusion

Silencing lncRNA KCNQ1OT1 can alleviate OGD/R-induced microglia injury via upregulating the expression of miR-145-5p and targeting the down-regulation of ROCK1 expression.

关键词

Keywords

references

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