Universidad de Buenos Aires, Facultad de Farmacia y Bioquímica, IQUIFIB (UBA-CONICET), Buenos Aires, Argentina.
INBIRS, Universidad de Buenos Aires (UBA-CONICET), Buenos Aires, Argentina.
Biochem Pharmacol. 2024 Nov;229:116480. doi: 10.1016/j.bcp.2024.116480. Epub 2024 Aug 10.
Alamandine (ALA) exerts protective effects similar to angiotensin (Ang) (1-7) through Mas-related G protein-coupled receptor type D receptor (MrgDR) activation, distinct from Mas receptor (MasR). ALA induces anti-inflammatory effects in mice but its impact in human macrophages remains unclear. We aimed to investigate the anti-inflammatory effects of ALA in human macrophages. Interleukin (IL)-6 and IL-1β were measured by ELISA in human THP-1 macrophages and human monocyte-derived macrophages exposed to lipopolysaccharide (LPS). Consequences of MasR-MrgDR heteromerization were investigated in transfected HEK293T cells. ALA decreased IL-6 and IL-1β secretion in LPS-activated THP-1 macrophages. The ALA-induced decrease in IL-6 but not in IL-1β was prevented by MasR blockade and MasR downregulation, suggesting MasR-MrgDR interaction. In human monocyte-derived M1 macrophages, ALA decreased IL-1β secretion independently of MasR. MasR-MrgDR interaction was confirmed in THP-1 macrophages, human monocyte-derived macrophages, and transfected HEK293T cells. MasR and MrgDR formed a constitutive heteromer that was not influenced by ALA. ALA promoted Akt and ERK1/2 activation only in cells expressing MasR-MrgDR heteromers, and this effect was prevented by MasR blockade. While Ang-(1-7) reduced cellular proliferation in MasR -but not MrgDR- expressing cells, ALA antiproliferative effect was elicited in cells expressing MasR-MrgDR heteromers. ALA also induced an antiproliferative response in THP-1 cells and this effect was abolished by MasR blockade, reinforcing MasR-MrgDR interaction. MasR-MrgDR heteromerization is crucial for ALA-induced anti-inflammatory and antiproliferative responses in human macrophages. This study broaden our knowledge of the protective axis of the RAS, thus enabling novel therapeutic approaches in inflammatory-associated diseases.
阿拉曼定 (ALA) 通过 Mas 相关 G 蛋白偶联受体 D 型受体 (MrgDR) 的激活发挥类似于血管紧张素 (Ang) (1-7) 的保护作用，与 Mas 受体 (MasR) 不同。ALA 在小鼠中诱导抗炎作用，但在人类巨噬细胞中的影响尚不清楚。我们旨在研究 ALA 在人类巨噬细胞中的抗炎作用。通过 ELISA 在人 THP-1 巨噬细胞和人单核细胞衍生的巨噬细胞中测量白细胞介素 (IL)-6 和 IL-1β，这些细胞暴露于脂多糖 (LPS)。在转染的 HEK293T 细胞中研究了 MasR-MrgDR 异源二聚体的后果。ALA 降低 LPS 激活的 THP-1 巨噬细胞中 IL-6 和 IL-1β 的分泌。ALA 诱导的 IL-6 减少，但不是 IL-1β 的减少，被 MasR 阻断和 MasR 下调所阻止，表明 MasR-MrgDR 相互作用。在人单核细胞衍生的 M1 巨噬细胞中，ALA 独立于 MasR 降低 IL-1β 的分泌。在 THP-1 巨噬细胞、人单核细胞衍生的巨噬细胞和转染的 HEK293T 细胞中证实了 MasR-MrgDR 相互作用。MasR 和 MrgDR 形成一种组成型异源二聚体，不受 ALA 的影响。ALA 仅在表达 MasR-MrgDR 异源二聚体的细胞中促进 Akt 和 ERK1/2 的激活，并且这种作用被 MasR 阻断所阻止。虽然 Ang-(1-7) 降低了表达 MasR 的细胞中的细胞增殖，但在表达 MasR-MrgDR 异源二聚体的细胞中，ALA 的抗增殖作用被引发。ALA 还在 THP-1 细胞中诱导抗增殖反应，这种作用被 MasR 阻断所消除，从而加强了 MasR-MrgDR 相互作用。MasR-MrgDR 异源二聚体对于 ALA 诱导的人类巨噬细胞抗炎和抗增殖反应至关重要。这项研究拓宽了我们对 RAS 保护轴的认识，从而为炎症相关疾病提供了新的治疗方法。
