A protective role of nintedanib in peritoneal fibrosis through H19–EZH2–KLF2 axis via impeding mesothelial-to-mesenchymal transition,International Urology and Nephrology

当前位置： X-MOL 学术 › Int. Urol. Nephrol. › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

A protective role of nintedanib in peritoneal fibrosis through H19–EZH2–KLF2 axis via impeding mesothelial-to-mesenchymal transition
International Urology and Nephrology ( IF 1.8 ) Pub Date : 2023-12-14 , DOI: 10.1007/s11255-023-03892-8
Wei Zhong ₁ , Jia Fu ₂ , Jin Liao ₁ , Shaxi Ouyang ₁ , Wei Yin ₁ , Yumei Liang ₁ , Kanghan Liu ₁

Affiliation

Background

Peritoneal fibrosis (PF), a common complication of long-term peritoneal dialysis, accounts for peritoneal ultrafiltration failure to develop into increased mortality. Nintedanib has previously been shown to protect against multi-organ fibrosis, including PF. Unfortunately, the precise molecular mechanism underlying nintedanib in the pathogenesis of PF remains elusive.

Methods

The mouse model of PF was generated by chlorhexidine gluconate (CG) injection with or without nintedanib administration, either with the simulation for the cell model of PF by constructing high-glucose (HG)-treated human peritoneal mesothelial cells (HPMCs). HE and Masson staining were applied to assess the histopathological changes of peritoneum and collagen deposition. FISH, RT-qPCR, western blot and immunofluorescence were employed to examine distribution or expression of targeted genes. Cell viability was detected using CCK-8 assay. Cell morphology was observed under a microscope. RNA immunoprecipitation (RIP) and chromatin immunoprecipitation (ChIP) assays were applied to validate the H19–EZH2–KLF2 regulatory axis.

Results

Aberrantly overexpressed H19 was observed in both the mouse and cell model of PF, of which knockdown significantly blocked HG-induced mesothelial-to-mesenchymal transition (MMT) of HPMCs. Moreover, loss of H19 further strengthened nintedanib-mediated suppressive effects against MMT process in a mouse model of PF. Mechanistically, H19 could epigenetically repressed KLF2 via recruiting EZH2. Furthermore, TGF-β/Smad pathway was inactivated by nintedanib through mediating H19/KLF2 axis.

Conclusion

In summary, nintedanib disrupts MMT process through regulating H19/EZH2/KLF2 axis and TGF-β/Smad pathway, which laid the experimental foundation for nintedanib in the treatment of PF.

中文翻译：

尼达尼布通过 H19-EZH2-KLF2 轴阻止间皮间质转化对腹膜纤维化的保护作用

背景

腹膜纤维化（PF）是长期腹膜透析的常见并发症，是腹膜超滤失败导致死亡率增加的原因。尼达尼布此前已被证明可以预防多器官纤维化，包括肺纤维化。不幸的是，尼达尼布在 PF 发病机制中的精确分子机制仍然难以捉摸。

方法

通过注射葡萄糖酸氯己定（CG）并给予或不给予尼达尼布来产生PF小鼠模型，或者通过构建高葡萄糖（HG）处理的人腹膜间皮细胞（HPMC）来模拟PF的细胞模型。应用HE和Masson染色评估腹膜和胶原沉积的组织病理学变化。采用FISH、RT-qPCR、蛋白质印迹和免疫荧光来检查目标基因的分布或表达。使用CCK-8测定法检测细胞活力。在显微镜下观察细胞形态。应用 RNA 免疫沉淀 (RIP) 和染色质免疫沉淀 (ChIP) 测定来验证 H19-EZH2-KLF2 调控轴。

结果

在 PF 小鼠和细胞模型中均观察到 H19 异常过度表达，其中敲低显着阻断 HG 诱导的 HPMC 间皮间质转化 (MMT)。此外，H19的缺失进一步增强了PF小鼠模型中尼达尼布介导的对MMT过程的抑制作用。从机制上讲，H19 可以通过招募 EZH2 来表观遗传抑制 KLF2。此外，尼达尼布通过介导 H19/KLF2 轴使 TGF-β/Smad 通路失活。