[1]王兴健,周巧艳,颜伟健,等.长链非编码lnc-FAF通过调控微小RNA-302a-3p保护缺氧复氧诱导肾小管上皮细胞的损伤[J].陕西医学杂志,2025,54(4):452-457.[doi:DOI:10.3969/j.issn.1000-7377.2025.04.004]
　WANG Xingjian,ZHOU Qiaoyan,YAN Weijian,et al.Lnc-FAF protects renal tubular epithelial cells from hypoxia-reoxygenation-induced injury by regulating miR-302a-3p[J].,2025,54(4):452-457.[doi:DOI:10.3969/j.issn.1000-7377.2025.04.004]
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长链非编码lnc-FAF通过调控微小RNA-302a-3p保护缺氧复氧诱导肾小管上皮细胞的损伤
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《陕西医学杂志》[ISSN:1000-7377/CN:61-1281/TN]

卷:
54
期数:
2025年4期
页码:
452-457
栏目:
基础研究
出版日期:
2025-04-05

文章信息/Info

Title:
Lnc-FAF protects renal tubular epithelial cells from hypoxia-reoxygenation-induced injury by regulating miR-302a-3p
作者:
王兴健1; 周巧艳1; 颜伟健1; 胡 超2
(1.邵阳学院附属第二医院肾内风湿免疫科,湖南 邵阳 422099; 2.复旦大学附属华山医院肾内科,上海 200040)
Author(s):
WANG Xingjian1; ZHOU Qiaoyan1; YAN Weijian1; HU Chao2
(1.Department of Nephrology and Rheumatology,The Second Affiliated Hospital of Shaoyang University,Shaoyang 422099,China; 2.Department of Nephrology,Huashan Hospital,Fudan University,Shanghai 200040,China)
关键词:
长链非编码RNA;  lnc-FAF;  肾小管上皮细胞;  微小RNA-302a-3p;  缺氧复氧;  细胞凋亡
Keywords:
Long noncoding RNA;  lnc-FAF;  Renal tubular epithelial cells;  MiR-302a-3p;  Hypoxia-reoxygenation;  Cell apoptosis
分类号:
R 692.6
DOI:
DOI:10.3969/j.issn.1000-7377.2025.04.004
文献标志码:
A
摘要:
目的:探讨长链非编码(lncRNA)lnc-FAF对缺氧复氧诱导肾小管上皮细胞损伤的影响及调控机制。方法:对HK-2细胞进行缺氧复氧处理,建立HK-2细胞损伤模型,实时定量聚合酶链反应(qRT-PCR)检测HK-2细胞中lnc-FAF表达。采用慢病毒感染技术上调HK-2细胞中lnc-FAF表达。细胞计数试剂盒(CCK-8)实验和流式细胞术分别检测各组HK-2细胞的活力和凋亡水平。酶联免疫吸附测定法(ELISA)检测各组HK-2细胞白细胞介素-1β(L-1β)、肿瘤坏死因子-α(TNF-α)和白细胞介素-6(IL-6)表达水平。Western blot法检测各组HK-2细胞中增殖表型细胞周期蛋白依赖性激酶2(Cyclin A)、细胞周期蛋白依赖性激酶2(CDK2)、促凋亡蛋白半胱氨酸天冬氨酸蛋白酶3(Caspase-3)、半胱氨酸天冬氨酸蛋白酶9(Caspase-9)表达。双荧光素酶报告实验验证lnc-FAF与微小RNA-302a-3p(miR-302a-3p)的靶向关系。qRT-PCR检测各组HK-2细胞中miR-302a-3p表达。结果:缺氧复氧诱导的HK-2细胞中lnc-FAF表达明显低于正常HK-2细胞(P<0.01)。lnc-FAF组HK-2细胞中lnc-FAF表达显著高于Control组(P<0.01)。lnc-FAF组HK-2细胞活力明显高于Control组(P<0.01),并且lnc-FAF组HK-2细胞凋亡率显著低于Control组(P<0.01)。与Control组相比,lnc-FAF组TNF-α、IL-1β和IL-6表达均明显下降(均P<0.01)。与Control组相比,lnc-FAF组HK-2细胞中增殖表型蛋白Cyclin A、CDK2表达均显著升高(均P<0.01),促凋亡蛋白Caspase-3、Caspase-9表达均明显降低(均P<0.01)。lnc-FAF可靶向结合miR-302a-3p(P<0.01)。lnc-FAF组HK-2细胞中miR-302a-3p表达明显低于Control组(P<0.01)。结论:lnc-FAF可靶向下调miR-302a-3p表达,减少缺氧复氧诱导的炎性因子,增强肾小管上皮细胞活力,抑制肾小管上皮细胞凋亡,减轻缺氧复氧诱导的肾小管上皮细胞损伤。
Abstract:
Objective:To investigate the effect and regulatory mechanism of long non-coding RNA(lncRNA)lnc-FAF on hypoxia-reoxygenation-induced renal tubular epithelial cell injury.Methods:HK-2 cells were treated with hypoxia and reoxygenation to establish a HK-2 cell injury model.Real-time quantitative polymerase chain reaction(qRT-PCR)was used to detect the expression of lnc-FAF in HK-2 cells.Lentivirus infection technology was used to upregulate the expression of lnc-FAF in HK-2 cells.CCK-8 assay and flow cytometry were used to detect the viability and apoptosis levels of HK-2 cells in each group,respectively.ELISA was used to detect the expression levels of interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)in HK-2 cells in each group.Western blot was used to detect the expression of proliferation phenotype Cyclin A,Cyclin-Dependent Kinase 2(CDK2)and pro-apoptotic proteins Cysteine-aspartic acid Protease 3(Caspase-3),Cysteine-aspartic acid Protease 9(Caspase-9)in HK-2 cells in each group.Dual luciferase reporter assay verified the targeting relationship between lnc-FAF and miR-302a-3p.qRT-PCR was used to detect the expression of miR-302a-3p in HK-2 cells of each group.Results:The expression of lnc-FAF in HK-2 cells induced by hypoxia and reoxygenation was significantly lower than that in normal HK-2 cells(P<0.01).The expression of lnc-FAF in HK-2 cells in the lnc-FAF group was significantly higher than that in the control group(P<0.01).The viability of HK-2 cells in the lnc-FAF group was significantly higher than that in the control group(P<0.01),and the apoptosis rate of HK-2 cells in the lnc-FAF group was significantly lower than that in the control group(P<0.01).Compared with the control group,the expressions of TNF-α,IL-1β and IL-6 in the lnc-FAF group were significantly decreased(all P<0.01).Compared with the control group,the expressions of proliferation phenotype proteins Cyclin A and CDK2 in HK-2 cells in the lnc-FAF group were significantly increased(all P<0.01),and the expressions of pro-apoptotic proteins Caspase-3 and Caspase-9 were significantly decreased(all P<0.01).lnc-FAF can target and bind to miR-302a-3p(P<0.01).The expression of miR-302a-3p in HK-2 cells in the lnc-FAF group was significantly lower than that in the control group(P<0.01).Conclusion:Lnc-FAF can target and downregulate the expression of miR-302a-3p,reduce inflammatory factors induced by hypoxia and reoxygenation,enhance the viability of renal tubular epithelial cells,inhibit the apoptosis of renal tubular epithelial cells,and alleviate the damage of renal tubular epithelial cells induced by hypoxia and reoxygenation.

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备注/Memo

备注/Memo:
[基金项目]国家自然科学基金资助项目(81900682)
更新日期/Last Update: 2025-04-07