Wang Yinglei, Li Yuansheng, Ni Dan, Wei Ziqi, Fu Zhe, Li Chao, Sun Huiling, Wu Yutong, Li Yilin, Zhang Yingxuan, Liu Naixin, Liu Yixiang, Wang Zhuo, Li Jiayi, Sun Dandan, He Li, Yang Ying, Wang Ying, Yang Xinwang
Department of Anatomy and Histology & Embryology, Faculty of Basic Medical Science, Kunming Medical University, Kunming, China.
Key Laboratory of Chemistry in Ethnic Medicinal Resources & Key Laboratory of Natural Products Synthetic Biology of Ethnic Medicinal Endophytes, State Ethnic Affairs Commission & Ministry of Education, School of Ethnic Medicine, Yunnan Minzu University, Kunming, China.
Environ Toxicol. 2023 Dec;38(12):2826-2835. doi: 10.1002/tox.23914. Epub 2023 Aug 11.
Active peptides play a vital role in the development of new drugs and the identification and discovery of drug targets. As the first reported native peptide homodimer with pro-regenerative potency, OA-GP11d could potentially be used as a novel molecular probe to help elucidate the molecular mechanism of skin wound repair and provide new drug targets.
Bioinformatics analysis and luciferase assay were adopted to determine microRNAs (miRNAs) and its target. The prohealing potency of the miRNA was determined by MTS and a Transwell experiment against mouse macrophages. Enzyme-linked immunosorbent assay, realtime polymerase chain reaction, and western blotting were performed to explore the molecular mechanisms.
In this study, OA-GP11d was shown to induce Mus musculus microRNA-186-5p (mmu-miR-186-5p) down-regulation. Results showed that miR-186-5p had a negative effect on macrophage migration and proliferation as well as a targeted and negative effect on TGF-β type II receptor (TGFβR2) expression and an inhibitory effect on activation of the downstream SMAD family member 2 (Smad2) and protein-p38 kinase signaling pathways. Importantly, delivery of a miR-186-5p mimic delayed skin wound healing in mice.
miR-186-5p regulated macrophage migration and proliferation to delay wound healing through the TGFβR2/Smad2/p38 molecular axes, thus providing a promising new pro-repair drug target.
活性肽在新药研发以及药物靶点的识别与发现中发挥着至关重要的作用。作为首个被报道的具有促再生潜能的天然肽同二聚体，OA-GP11d 有可能被用作一种新型分子探针，以帮助阐明皮肤伤口修复的分子机制并提供新的药物靶点。
采用生物信息学分析和荧光素酶测定法来确定微小RNA（miRNA）及其靶点。通过MTS法和针对小鼠巨噬细胞的Transwell实验来测定miRNA的促愈合潜能。进行酶联免疫吸附测定、实时聚合酶链反应和蛋白质印迹法以探究分子机制。
在本研究中，OA-GP11d 被证明可诱导小家鼠微小RNA-186-5p（mmu-miR-186-5p）表达下调。结果表明，miR-186-5p 对巨噬细胞迁移和增殖具有负面影响，对转化生长因子-βⅡ型受体（TGFβR2）表达具有靶向性负面影响，并对下游SMAD家族成员2（Smad2）的激活以及蛋白p38激酶信号通路具有抑制作用。重要的是，给予miR-186-5p模拟物会延迟小鼠皮肤伤口愈合。
miR-186-5p 通过TGFβR2/Smad2/p38分子轴调节巨噬细胞迁移和增殖，从而延迟伤口愈合，因此提供了一个有前景的新的促修复药物靶点。
