Guangxi Universities Key Laboratory of Stem Cell and Biopharmaceutical Technology, Guangxi Normal University, Guangxi Zhuang Autonomous Region, Guilin, China; Research Center for Biomedical Sciences, Guangxi Normal University, Guangxi Zhuang Autonomous Region, Guilin, China; College of Life Sciences, Guangxi Normal University, Guangxi Zhuang Autonomous Region, Guilin, China.
Department of Biomedical Sciences, College of Veterinary Medicine and Life Science, City University of Hong Kong, Hong Kong SAR, China.
J Ethnopharmacol. 2024 Sep 15;331:118272. doi: 10.1016/j.jep.2024.118272. Epub 2024 May 6.
Rehmanniae Radix Praeparata (RRP), a staple in traditional Chinese medicine, is derived from Rehmannia glutinosa Libosch and is renowned for its wound-healing properties. Despite its clinical prevalence, the molecular mechanisms underlying RRP's wound-healing effects have not been fully elucidated.
This research endeavored to delineate the molecular and cellular mechanisms underlying the beneficial effects of RRP on wound healing, utilizing a zebrafish model.
Zebrafish larvae at 3 days post-fertilization were amputated at the fin and subsequently treated with RRP. The pro-wound healing and regenerative effects of RRP were evaluated through morphological analysis, assessment of cell proliferation and apoptosis, Additionally, mechanistic insights were gained through a comprehensive approach encompassing network pharmacology analysis, cell tracing, RNA-sequencing, CRISPR/Cas9 gene editing, and pharmacological inhibition.
Our findings demonstrate that RRP significantly accelerates caudal fin regeneration in zebrafish following injury by suppressing cell apoptosis, promoting cell proliferation, and upregulating the expression of regenerative-related genes. Furthermore, RRP triggers autophagy signals during the regenerative process, which is attenuated by the autophagy inhibitor chloroquine (CQ). Notably, the administration of RRP enhances the expression of ahr1 and ahr2 in the regenerating fin. Genetic knockout of ahr1a, ahr1b, or ahr2 using CRISPR/Cas9, or pharmacological blockade of AHR signals with the antagonist CH-223191, diminishes the regenerative potential of RRP. Remarkably, zebrafish lacking ahr2 completely lose their fin regeneration ability. Additionally, inhibition of AHR signaling suppresses autophagy signaling during fin regeneration.
This study uncovers that RRP stimulates fin regeneration in zebrafish by inducing AHR signals and, at least partially, activating the autophagy process. These findings provide novel insights into the molecular mechanisms underlying the wound-healing effects of RRP and may pave the way for the development of novel therapeutic strategies.
标题 民族药理学相关性：熟地黄（RRP）是中药的主要成分之一，来源于地黄，以其愈合伤口的特性而闻名。尽管在临床上广泛应用，但 RRP 促进伤口愈合的确切分子机制尚未完全阐明。
本研究旨在利用斑马鱼模型阐明 RRP 促进伤口愈合的有益作用的分子和细胞机制。
受精后 3 天的斑马鱼幼鱼在鳍处截肢，然后用 RRP 处理。通过形态分析、细胞增殖和凋亡评估，评估 RRP 的促伤口愈合和再生作用。此外，通过网络药理学分析、细胞示踪、RNA 测序、CRISPR/Cas9 基因编辑和药理学抑制等综合方法，获得了机制见解。
我们的研究结果表明，RRP 通过抑制细胞凋亡、促进细胞增殖和上调再生相关基因的表达，显著加速了斑马鱼受伤后的尾鳍再生。此外，RRP 在再生过程中触发自噬信号，自噬抑制剂氯喹（CQ）可减弱该信号。值得注意的是，RRP 给药可增强再生鳍中的 ahr1 和 ahr2 的表达。使用 CRISPR/Cas9 对 ahr1a、ahr1b 或 ahr2 进行基因敲除，或用拮抗剂 CH-223191 阻断 AHR 信号，均会降低 RRP 的再生潜力。值得注意的是，缺乏 ahr2 的斑马鱼完全丧失了鳍的再生能力。此外，抑制 AHR 信号会抑制鳍再生过程中的自噬信号。
本研究揭示 RRP 通过诱导 AHR 信号，至少部分激活自噬过程，刺激斑马鱼的鳍再生。这些发现为 RRP 促进伤口愈合的确切分子机制提供了新的见解，并可能为开发新的治疗策略铺平道路。
