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  

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 颜伟健[1] 胡超 WANG Xingjian;ZHOU Qiaoyan;YAN Weijian;HU Chao(Department of Nephrology and Rheumatology,The Second Affiliated Hospital of Shaoyang University,Shaoyang 422099,China;Department of Nephrology,Huashan Hospital,Fudan University,Shanghai 200040,China)

机构地区：[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

出　　处：《陕西医学杂志》2025年第4期452-457,共6页Shaanxi Medical Journal

基　　金：国家自然科学基金资助项目(81900682)。

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。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-1βand 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 o

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 

分 类 号：R692.6[医药卫生—泌尿科学]