Scholarlink

Objective <p>To investigate the role of leukotriene B4 receptor 1 (LTB4R1) in high glucose-induced renal tubular epithelial cell injury and its regulatory association with macrophage polarization, and to validate the protective effect of LTB4R1 silencing in diabetic renal injury in vivo.</p> Methods <p>THP-1 cells were differentiated and polarized into M0, M1 (LPS + IFN-γ-induced) and M2 (IL-4 + IL-13-induced) macrophages, with phenotypic validation by immunofluorescence. HK-2 renal tubular epithelial cells were treated with gradient glucose concentrations (5.5, 15, 30, 45 mM) to assess dose-dependent effects on LTB4R1 expression and apoptosis. Indirect co-culture of HK-2 cells with polarized macrophages under high glucose (HG) conditions was performed using a Transwell system, with an osmotic control group to exclude osmotic artifacts. For in vivo studies, STZ-induced diabetic C57BL/6 mice were treated with AAV-mediated LTB4R1 shRNA. qPCR, Western blot, ELISA and flow cytometry were used to detect molecular expression, inflammatory cytokines and apoptosis.</p> Results <p>THP-1 cells were successfully differentiated into M0, M1 and M2 macrophages with distinct phenotypic marker expression. High glucose dose-dependently upregulated LTB4R1 mRNA and protein levels (all <i>P</i> &lt; 0.01) and promoted HK-2 cell apoptosis (<i>P</i> &lt; 0.01). HG specifically induced HK-2 cell injury, which was exacerbated by M1 macrophages and alleviated by M2 macrophages; concurrent changes included HG-induced upregulation of LTB4R1/CD68 and downregulation of CD163/CD206, which were reversed by M2 co-culture (all <i>P</i> &lt; 0.01). In diabetic mice, LTB4R1 silencing significantly reduced renal Ltb4r1, cleaved caspase-3 and NF-κB p65 transcripts, suppressed pro-inflammatory cytokines (TNF-α, IL-1β) and elevated anti-inflammatory IL-10, restored CD163/CD206 expression, and inhibited renal apoptosis (all <i>P</i> &lt; 0.01).</p> Conclusion <p>LTB4R1 mediates high glucose-induced renal tubular epithelial cell injury and diabetic renal injury by regulating macrophage polarization and NF-κB signaling. Silencing LTB4R1 mitigates diabetic renal inflammation and apoptosis, suggesting LTB4R1 as a potential therapeutic target for diabetic nephropathy that warrants further investigation.</p>

错误：搜索内容不能为空，请输入英文关键词
错误：关键词超出字数限制，请精简
高级检索

Mechanisms by which macrophage Ltb4r1 mediates tubular epithelial cell injury to facilitate the advancement of diabetic kidney disease

  • 期刊论文

  • 出版模式：

    开放获取

  • 发表日期：

    2026年7月8日

查看全文
  • Wenjing Cao,
  • Qingzhen Liu,
  • Natalia Viktorovna Chebotareva,
  • Shasha Lv,
  • Jing Cheng,
  • Shanshan Liu,
  • Fulin Dou,
  • Gang Liu

摘要

Objective

To investigate the role of leukotriene B4 receptor 1 (LTB4R1) in high glucose-induced renal tubular epithelial cell injury and its regulatory association with macrophage polarization, and to validate the protective effect of LTB4R1 silencing in diabetic renal injury in vivo.

Methods

THP-1 cells were differentiated and polarized into M0, M1 (LPS + IFN-γ-induced) and M2 (IL-4 + IL-13-induced) macrophages, with phenotypic validation by immunofluorescence. HK-2 renal tubular epithelial cells were treated with gradient glucose concentrations (5.5, 15, 30, 45 mM) to assess dose-dependent effects on LTB4R1 expression and apoptosis. Indirect co-culture of HK-2 cells with polarized macrophages under high glucose (HG) conditions was performed using a Transwell system, with an osmotic control group to exclude osmotic artifacts. For in vivo studies, STZ-induced diabetic C57BL/6 mice were treated with AAV-mediated LTB4R1 shRNA. qPCR, Western blot, ELISA and flow cytometry were used to detect molecular expression, inflammatory cytokines and apoptosis.

Results

THP-1 cells were successfully differentiated into M0, M1 and M2 macrophages with distinct phenotypic marker expression. High glucose dose-dependently upregulated LTB4R1 mRNA and protein levels (all P < 0.01) and promoted HK-2 cell apoptosis (P < 0.01). HG specifically induced HK-2 cell injury, which was exacerbated by M1 macrophages and alleviated by M2 macrophages; concurrent changes included HG-induced upregulation of LTB4R1/CD68 and downregulation of CD163/CD206, which were reversed by M2 co-culture (all P < 0.01). In diabetic mice, LTB4R1 silencing significantly reduced renal Ltb4r1, cleaved caspase-3 and NF-κB p65 transcripts, suppressed pro-inflammatory cytokines (TNF-α, IL-1β) and elevated anti-inflammatory IL-10, restored CD163/CD206 expression, and inhibited renal apoptosis (all P < 0.01).

Conclusion

LTB4R1 mediates high glucose-induced renal tubular epithelial cell injury and diabetic renal injury by regulating macrophage polarization and NF-κB signaling. Silencing LTB4R1 mitigates diabetic renal inflammation and apoptosis, suggesting LTB4R1 as a potential therapeutic target for diabetic nephropathy that warrants further investigation.