The total iridoid glycoside extract of Lamiophlomis rotata Kudo induces M2 macrophage polarization to accelerate wound healing by RAS/ p38 MAPK/NF-κB pathway.
The total iridoid glycoside extract of Lamiophlomis rotata Kudo induces M2 macrophage polarization to accelerate wound healing by RAS/ p38 MAPK/NF-κB pathway.
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DOI：
10.1016/j.jep.2023.116193
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发表时间：
2023-02
影响因子：
5.4
通讯作者：
Lei Lei-Lei;Guoguo Wan;Xiaoyu Geng;Jianguo Sun;Y. Zhang;Jianwei Wang;Congwen Yang;Zheng Pan
中科院分区：
文献类型：
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作者：
Lei Lei-Lei;Guoguo Wan;Xiaoyu Geng;Jianguo Sun;Y. Zhang;Jianwei Wang;Congwen Yang;Zheng Pan
民族药理学相关性独一味Lamiophlomis rotata（Benth.）苦藤（L.rotata），一种西藏药用植物，用于治疗“黄水病”，如皮肤病、黄疸和风湿病。本实验室前期研究表明，L. Rotata（IGLR）是皮肤伤口愈合的主要成分。然而，IGLR在创伤修复的生物学过程中的作用和可能的作用机制在很大程度上仍然未知。免疫荧光染色观察IGLR的抗炎、促血管生成和纤维形成作用，实时荧光定量PCR（RT-PCR）和酶联免疫吸附试验（ELISA）检测促炎因子和生长因子的表达。培养巨噬细胞、真皮成纤维细胞和内皮细胞，检测IGLR对细胞增殖和迁移的直接/间接相互作用，并采用流式细胞术检测IGLR对巨噬细胞表型的影响。结果IGLR可通过RAS/p38 MAPK/NF-κB信号通路，促进在体和离体创伤中M2标志物CD 206的表达。IGLR在创伤愈合早期抑制炎症细胞因子iNOS、IL-1β和TNF-α的表达，在创伤修复的增殖阶段，IGLR通过增加VEGF、CD 31、TGF-β和α-SMA的表达促进创伤组织血管新生和纤维形成，RT-PCR和ELISA结果证实了IGLR的作用。在旁分泌机制中，提取物促进真皮成纤维细胞的增殖，和内皮细胞的条件培养基（CM）成立。结论IGLR诱导M2巨噬细胞极化在伤口愈合的早期阶段，反过来，IGLR发挥了关键作用，在伤口愈合的生物过程中从炎症到细胞增殖的过渡。
Ethnopharmacological relevanceLamiophlomis rotata(Benth.)Kudo(L.rotata), a Tibetan medicinal plant, is used to treat “yellow-water diseases”, such as skin disease, jaundice and rheumatism. Our previous study showed that the iridoid glycoside extract ofL. rotata(IGLR) is the major constituent of skin wound healing. However, the role of IGLR in the biological process of trauma repair and the probable mechanism of the action remain largely unknown.Aim of the studyTo investigate the role of IGLR in the biological process of trauma repair and the probable mechanism of the action.Materials and methodsThe role of IGLR in wound healing was investigated by overall skin wound in mice with Hematoxylin and Eosin (H&E) and Masson trichrome staining. The anti-inflammatory, angiogenesis-promoting and fibril formation effects of IGLR were visualized in wound skin tissue by immunofluorescence staining, and the proinflammatory factors and growth factors were assayed by real-time polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA). Macrophages, dermal fibroblasts, and endothelial cells were cultured to measure the direct/indirect interaction effects of IGLR on the proliferation and migration of cells, and flow cytometry was employed to assess the role of IGLR on macrophage phenotype. Network pharmacology combined with Western blot experiments were conducted to explore possible mechanisms of the actions.ResultsIGLR increased the expression of CD206 (M2 markers) through the RAS/p38 MAPK/NF-κB signaling pathway during wound injuryin vivoandin vitro. IGLR suppressed the inflammatory cytokines iNOS, IL-1β and TNF-α in the early stage of wound healing.During the proliferation step of wound repair, IGLR promoted angiogenesis and fibril formation by increasing the expression of VEGF, CD31, TGF-β and α-SMA in wound tissue, and similar results were verified by RT-PCR and ELISA. In a paracrine mechanism, the extract promoted the proliferation of dermal fibroblasts, and endothelial cells were founded by the conditioned medium (CM).ConclusionIGLR induced M2 macrophage polarization in the early stage of wound healing; in turn, IGLR played a key role in the transition from inflammation to cell proliferation during the biological process of wound healing.