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目的探讨电压依赖性阴离子通道蛋白2(VDAC2)在铁死亡调控肺纤维化(PF)中的作用及其与微管聚合促进蛋白3(TPPP3)的关系。方法将人非小细胞肺癌A549细胞随机分为对照组(C组)、铁死亡诱导剂Erastin处理组(E组)、转化生长因子β?(TGF-β?)联合Erastin处理组(T+E组)、VDAC2基因沉默联合Erastin处理组(shVDAC2+E组)。采用CCK-8法检测细胞活性，实时荧光定量聚合酶链反应(RT-qPCR)检测VDAC2、TPPP3、E-钙黏蛋白(E-cad)、α-平滑肌肌动蛋白(α-SMA)、谷胱甘肽过氧化物酶4(GPX4)mRNA，比色法检测亚铁离子(Fe2?)，免疫荧光技术检测VDAC2、TPPP3蛋白，荧光探针法检测活性氧(ROS)，酶联免疫吸附试验检测4-羟基壬烯醛(4-HNE)。采用Pearson相关法分析VDAC2与TPPP3表达的相关性。结果与C组比较，E组、T+E组细胞活性低(P均<0.05)，VDAC2、TPPP3mRNA和蛋白表达高(P均<0.05)，α-SMAmRNA表达高、E-cadmRNA表达低(P均<0.05)，GPX4mRNA表达低(P均<0.05)，Fe2+、ROS、4-HNE水平高(P均<0.05)。与E组比较，T+E组上述变化更显著(P均<0.05)，而shVDAC2+E组细胞活性高(P均<0.05)，VDAC2、TPPP3mRNA和蛋白表达低(P均<0.05)，α-SMAmRNA表达低、E-cadmRNA表达高(P均<0.05)，GPX4mRNA表达高(P<0.05)，Fe2+、ROS、4-HNE水平低(P均<0.05)。Pearson相关性分析显示，VDAC2与TPPP3的mRNA及蛋白表达均呈正相关(r分别为0.980、0.984，P均<0.05)。结论VDAC2在Erastin诱导的铁死亡中表达上调，并可能通过正向调控TPPP3促进铁死亡，进而加剧TGF-β?诱导的A549细胞EMT及纤维化进程；沉默VDAC2可降低TPPP3表达并逆转上述效应。
Abstract：Objective Investigating the regulatory role and mechanism of the mitochondrial transmembrane protein voltage-dependent anion channel 2 (VDAC2) in ferroptosis-mediated pulmonary fibrosis(PF) and its relationship with tubulin polymerization promoting protein 3 (TPPP3). Methods Human A549 cells were randomly divided into control group (group C)， ferroptosis inducer Erastin treatment group (group E)， transforming growth factor-β? (TGF-β?) combined with Erastin treatment group (group T+E)， and VDAC2 gene silencing combined with Erastin treatment group (group shVDAC2+E). Cell viability was assessed using the CCK-8 assay. The mRNA expression levels of VDAC2， TPPP3， E-cadherin (E-cad)， α-smooth muscle actin (α-SMA)， and glutathione peroxidase 4 (GPX4) were detected by real-time quantitative polymerase chain reaction (RT-qPCR). Ferrous ion (Fe?) levels were measured using a colorimetric assay. Immunofluorescence staining was employed to detect VDAC2 and TPPP3 proteins. Reactive oxygen species (ROS) were measured with a fluorescent probe. 4-Hydroxynonenal (4-HNE) levels were determined by enzyme-linked immunosorbent assay (ELISA). The correlation between VDAC2 and TPPP3 expression was analyzed using Pearson correlation analysis. Results Compared with group C， groups E and T+E showed lower cell viability(all P<0.05)， higher α-SMA mRNA expression and higher VDAC2 and TPPP3 mRNA and protein expression(all P<0.05)， lower E-cad and GPX4 mRNA expression(all P<0.05)， and higher levels of Fe?， ROS， and 4-HNE (all P<0.05). Compared with group E， the above changes were more pronounced in the group T+E (all P<0.05). In contrast， the group shVDAC2+E exhibited higher cell viability(all P<0.05)， lower VDAC2 and TPPP3 mRNA and protein expression and lower α-SMA mRNA expression(all P<0.05)， higher E-cad and GPX4 mRNA expression(all P<0.05)， and lower levels of Fe?， ROS， and 4-HNE (all P<0.05). Pearson correlation analysis revealed that VDAC2 expression was positively correlated with TPPP3 expression at both the mRNA and protein levels (r=0.980 and 0.984， respectively; both P<0.05). Conclusion VDAC2 is upregulated in Erastin-induced ferroptosis and promotes ferroptosis by positively regulating TPPP3， thereby exacerbating TGF-β?-induced epithelial-mesenchymal transition (EMT) and fibrotic progression in A549 cells. Silencing VDAC2 can reverse these effects.
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基本信息:
中图分类号:R563
引用信息:
[1]闫丽洁,张建,王士成,等.VDAC2在铁死亡调控肺纤维化中的作用及其与微管聚合促进蛋白3的关系[J].山东医药().
基金信息:
山东省自然科学基金项目（ZR2019MC019）
2026-08-05
2026-08-05
2026-08-05