Constitutive HO-1 and CD55 (DAF) Expression and Regulatory Interaction in Cultured Podocytes.
Constitutive HO-1 and CD55 (DAF) Expression and Regulatory Interaction in Cultured Podocytes.
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DOI：
10.3390/biomedicines11123297
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发表时间：
2023-12-13
期刊：
影响因子：
4.7
通讯作者：
中科院分区：
文献类型：
--
作者：
用体内转基因方法在内脏肾小球上皮细胞(足细胞)过表达可诱导的血红素加氧酶(HO-1)亚型，可增加免疫损伤大鼠肾小球补体调节蛋白衰变加速因子(DAF，CD55)的表达，减少补体的激活/沉积。在这项初步研究中，我们评估了结构性表达的HO-1是否调节培养的大鼠足细胞CD55的表达。我们采用流式细胞术、定量(Q)RT-qPCR和转录后HO-1基因沉默(HO-1干扰RNA，RNAi)的方法来检测培养的大鼠足细胞中足细胞HO-1和CD55mRNA的组成(基础)水平的变化。此外，我们还研究了HO-1诱导剂--血红素对HO-1和CD55表达的影响。结果表明，大鼠足细胞结构性地表达HO-1和DAF，并且HO-1的诱导剂血红素增加了HO-1和DAF的表达。用RNA干扰技术(RNAi)沉默HO-1基因是可行的，但对CD55转录的影响并不一致。这些观察结果与足细胞暴露于血红素的条件有关，该条件可激活补充级联反应，如全身或肾小球内溶血。
Overexpression of the inducible heme oxygenase (HO-1) isoform in visceral renal glomerular epithelial cells (podocytes) using in vivo transgenesis methods was shown to increase glomerular expression of the complement regulatory protein decay-accelerating factor (DAF, CD55) and reduce complement activation/deposition in a rat model of immune-mediated injury. In this preliminary study, we assessed whether constitutively expressed HO-1 regulates CD55 expression in cultured rat podocytes. We employed methods of flow cytometry, quantitative (q) RT-qPCR and post-transcriptional HO-1 gene silencing (HO-1 interfering RNA, RNAi), to assess changes in constitutive (basal) levels of podocyte HO-1 and CD55 mRNA in cultured rat podocytes. Additionally, the effect of the HO-1 inducer, heme, on HO-1 and CD55 expression was assessed. Results indicate that rat podocytes constitutively express HO-1 and DAF and that the HO-1 inducer, heme, increases both HO-1 and DAF expression. HO-1 gene silencing using RNA interference (RNAi) is feasible but the effect on constitutive CD55 transcription is inconsistent. These observations are relevant to conditions of podocyte exposure to heme that can activate the complementary cascade, as may occur in systemic or intraglomerular hemolysis.
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