■为了研究LCZ696,一种血管紧张素受体-脑啡肽抑制剂(ARNI)的作用和机制,腹膜间皮细胞的上皮-间质转化(EMT)和巨噬细胞M2极化。
■我们检查了LCZ696在4.25%高糖腹膜透析液(PDF)诱导的腹膜纤维化(PF)小鼠模型中的作用,并探讨了LCZ696对TGF-β1(5ng/mL)和IL-4(10ng/mL)刺激的Raw264.7细胞的作用机制。为了进一步阐明机制,我们用Raw264.7细胞的条件培养基处理HPMC。
■LCZ696有效改善了PDF小鼠的PF并抑制了EMT过程。体外,LCZ696还显著减轻TGF-β1诱导的HPMC的EMT,尽管与缬沙坦治疗组相比没有统计学上的显着差异。此外,LCZ696改善蜗牛和Slug的表达增加,驱动EMT的两个核转录因子。机械上,TGF-β1增加TGFβRI的表达,p-Smad3、p-PDGFRβ和p-EGFR,而用LCZ696治疗消除了TGF-β/Smad3,PDGFRβ和EGFR信号通路的激活。此外,Raw264.7暴露于IL-4导致精氨酸酶-1、CD163和p-STAT6的表达增加。用LCZ696处理通过使STAT6信号通路失活而抑制IL-4引起的M2巨噬细胞极化。此外,我们观察到LCZ696通过阻断M2巨噬细胞分泌TGF-β1抑制EMT。
■我们的研究表明,LCZ696通过阻断TGF-β/Smad3,PDGFRβ和EGFR通路,改善PF并改善TGF-β1诱导的HPMCEMT。同时,LCZ696还通过调节STAT6途径抑制M2巨噬细胞极化。
UNASSIGNED: To investigate the effects and mechanisms of LCZ696, an angiotensin receptor-neprilysin inhibitor (ARNI), on epithelial-mesenchymal transition (EMT) of peritoneal mesothelial cells and on macrophage M2 polarization.
UNASSIGNED: We examined the effects of LCZ696 in a 4.25% high glucose peritoneal dialysis fluid (PDF)-induced peritoneal fibrosis (PF) mouse model, and explored the mechanisms of LCZ696 on human peritoneal mesothelial cells (HPMCs) stimulated by TGF-β1 (5 ng/mL) and on Raw264.7 cells stimulated by IL-4 (10 ng/mL). To further elucidate the mechanism, we treated HPMCs with the conditioned medium of Raw264.7 cells.
UNASSIGNED: LCZ696 effectively improved PF and inhibited the process of EMT in PDF mice. In vitro, LCZ696 also significantly alleviated the EMT of TGF-β1 induced HPMCs, although there was no statistically significant difference when compared to the Valsartan treatment group. Moreover, LCZ696 ameliorates the increased expression of Snail and Slug, two nuclear transcription factors that drive the EMT. Mechanistically, TGF-β1 increased the expression of TGFβRI, p-Smad3, p-PDGFRβ and p-EGFR, while treatment with LCZ696 abrogated the activation of TGF-β/Smad3, PDGFRβ and EGFR signaling pathways. Additionally, exposure of Raw264.7 to IL-4 results in increasing expression of Arginase-1, CD163 and p-STAT6. Treatment with LCZ696 inhibited IL-4-elicited M2 macrophage polarization by inactivating the STAT6 signaling pathway. Furthermore, we observed that LCZ696 inhibits EMT by blocking TGF-β1 secretion from M2 macrophages.
UNASSIGNED: Our study demonstrated that LCZ696 improves PF and ameliorates TGF-β1-induced EMT of HPMCs by blocking TGF-β/Smad3, PDGFRβ and EGFR pathways. Meanwhile, LCZ696 also inhibits M2 macrophage polarization by regulating STAT6 pathway.