Lei Lei, Wan Guoguo, Geng Xiaoyu, Sun Jianguo, Zhang Yi, Wang Jianwei, Yang Congwen, Pan Zheng
Chongqing Medical University, Chongqing, China.
Chengdu University of Traditional Chinese Medicine, Chengdu 611130, China.
J Ethnopharmacol. 2023 May 10;307:116193. doi: 10.1016/j.jep.2023.116193. Epub 2023 Feb 4.
Lamiophlomis 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 of L. 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.
To investigate the role of IGLR in the biological process of trauma repair and the probable mechanism of the action.
The 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.
IGLR increased the expression of CD206 (M2 markers) through the RAS/p38 MAPK/NF-κB signaling pathway during wound injury in vivo and in 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).
IGLR 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.
藏药独一味（Lamiophlomis rotata (Benth.) Kudo）用于治疗“黄水病”，如皮肤病、黄疸和风湿病。我们之前的研究表明，独一味环烯醚萜苷提取物（IGLR）是皮肤伤口愈合的主要成分。然而，IGLR在创伤修复生物学过程中的作用及其可能的作用机制在很大程度上仍不清楚。
研究IGLR在创伤修复生物学过程中的作用及其可能的作用机制。
通过苏木精-伊红（H&E）染色和Masson三色染色观察IGLR对小鼠全层皮肤伤口愈合的作用。通过免疫荧光染色观察IGLR在伤口皮肤组织中的抗炎、促血管生成和纤维形成作用，通过实时聚合酶链反应（RT-PCR）和酶联免疫吸附测定（ELISA）检测促炎因子和生长因子。培养巨噬细胞、真皮成纤维细胞和内皮细胞，以测定IGLR对细胞增殖和迁移的直接/间接相互作用，采用流式细胞术评估IGLR对巨噬细胞表型的作用。进行网络药理学结合蛋白质免疫印迹实验以探索可能的作用机制。
在体内和体外伤口损伤过程中，IGLR通过RAS/p38丝裂原活化蛋白激酶/核因子-κB信号通路增加CD206（M2标志物）的表达。IGLR在伤口愈合早期抑制炎性细胞因子诱导型一氧化氮合酶（iNOS）、白细胞介素-1β（IL-1β）和肿瘤坏死因子-α（TNF-α）。在伤口修复的增殖阶段，IGLR通过增加伤口组织中血管内皮生长因子（VEGF）、CD31、转化生长因子-β（TGF-β）和α-平滑肌肌动蛋白（α-SMA）的表达促进血管生成和纤维形成，RT-PCR和ELISA验证了类似结果。在旁分泌机制中，提取物促进真皮成纤维细胞的增殖，条件培养基（CM）可促进内皮细胞增殖。
IGLR在伤口愈合早期诱导M2巨噬细胞极化；反过来，IGLR在伤口愈合生物学过程中从炎症向细胞增殖的转变中起关键作用。
