To investigate the effect of Angelica sinensis on thymic cortical regeneration in rapamycin-treated mice and its underlying mechanisms. The chemical components of Angelica sinensis were analyzed using high-resolution liquid chromatography-mass spectrometry (LC-MS) and ultra-performance liquid chromatography (UPLC). Seventy-two 6-8-week-old female BALB/c mice were randomly assigned by weight into six groups (n=12 per group): blank control, model control, normal regeneration, and Angelica sinensis small- (1 g/kg), medium- (2 g/kg), and large-dose (4 g/kg) groups. Except for the blank control group, acute thymic involution was induced in all other groups via intraperitoneal injection of rapamycin (1 mg·kg－1·d－1) for 3 consecutive days. After modeling, the Angelica sinensis-treated groups received oral gavage of corresponding doses for 7 days, while the normal regeneration and blank control groups received an equal volume of saline. Body weight and dorsal hair growth were recorded daily. Forelimb grip strength was measured 2 hours after the last administration. Thymic structure and the spatial distribution of thymic epithelial cells (TECs) and thymocytes were assessed by hematoxylin-eosin (HE) and immunofluorescence staining. Development and homeostasis of T-cell subsets in the thymus and peripheral blood were analyzed by flow cytometry and rapid Wright-Giemsa staining. T-cell receptor excision circles (TRECs) in genomic DNA from peripheral blood mononuclear cells were detected by quantitative PCR (qPCR). The mRNA expression levels of thymic function-related genes, inflammatory factors, and Wnt pathway-related genes were measured by quantitative reverse transcription PCR (qRT-PCR). Potential targets and pathways were screened by integrating network pharmacology prediction and molecular docking. Rapamycin successfully induced acute thymic atrophy. The model control group showed an approximately 50% decrease in thymic index (P<0.01), significantly weakened grip strength (P<0.05), and no obvious hair regeneration. Histologically, the thymic cortical area was reduced, with a blurred corticomedullary junction, disrupted continuity of the cortical TEC (cTEC) cytoplasmic process network, early-stage blockade of thymocyte development, disturbed spatial distribution, and inhibited thymic output. Compared with the normal regeneration group, medium and large doses of Angelica sinensis dose-dependently increased the cortical area, significantly enhanced the density of cortical CK8+ TECs and restored the continuity of their cytoplasmic process network (P<0.01), while upregulating the expression of Foxn1 (P<0.01) and its downstream target gene Dll4. Regarding thymocyte development, Angelica sinensis (large dose) significantly increased the proportion of CD3+ TCRβ+ thymocytes (P<0.05), promoted the balanced differentiation of CD4+CD8+ cells into mature single-positive (SP) thymocytes, and re-established normal spatial localization, manifested as increased density of CD8+ and CD4+CD8+ cells in the cortex and reaggregation of TCRβ+ cells in the medulla. Furthermore, Angelica sinensis (medium dose) significantly reduced the mRNA levels of thymic pro-inflammatory factors TNF-α, TGF-β, and IGFBP5 (P<0.05), thereby improving the local inflammatory microenvironment. For peripheral homeostasis, Angelica sinensis intervention maintained and increased the proportion of peripheral blood CD3+ T cells, elevated the percentage of CD34+ hematopoietic stem cells across all dosage groups (P<0.05), and upregulated the expression of the thymic homing factor Ccl25. Mechanistically, network pharmacology predicted the Wnt pathway as a potential target of Angelica sinensis active components. Experimental validation revealed that Angelica sinensis (medium dose) significantly upregulated Wnt4 mRNA expression, inhibited Gsk3β, and increased CTNNB1 (β-catenin) levels in the thymus (P<0.05), indicating activation of the Wnt/CTNNB1/Foxn1 signaling pathway to drive cTEC cytoskeletal repair and thymic regeneration. Angelica sinensis promotes cortical regeneration and functional recovery after rapamycin-induced acute thymic involution by activating the Wnt/CTNNB1/Foxn1 signaling pathway and improving the thymic inflammatory microenvironment, which collaboratively facilitate cortical thymic epithelial cell cytoskeletal repair and hematopoietic stem cell homing. This suggests its potential benefit for counteracting immune aging. 目的: 探讨当归促进雷帕霉素处理后的小鼠胸腺的皮质再生作用及其机制。方法: 采用高分辨液相色谱-质谱联用（LC-MS）、超高效液相色谱（UPLC）分析当归化学成分。6~8周龄雌性BALB/c小鼠72只，按体重随机分为空白对照组、模型对照组、正常再生组及当归小（1 g/kg）、中（2 g/kg）、大剂量组（4 g/kg），每组12只。除空白对照组外，其余各组通过腹腔注射雷帕霉素（1 mg·kg-1·d-1，连续3 d）建立急性胸腺退化模型。造模后，当归各剂量组连续灌胃给药7 d，正常再生组和空白对照组给予等量等渗氯化钠溶液。实验期间每日记录体重及背部毛发生长情况;于末次给药2 h后测定四肢抓力。采用苏木精-伊红（HE）染色和免疫荧光染色评估胸腺结构、胸腺上皮细胞（TEC）及胸腺细胞的空间分布;流式细胞术和快速瑞特-吉姆萨染色染色分析胸腺及外周血T细胞亚群发育及稳态;定量PCR检测小鼠外周血单个核细胞基因组DNA中T细胞受体切除环（TREC）水平;定量逆转录PCR检测小鼠胸腺功能、炎症因子和Wnt通路相关基因表达水平;结合网络药理学预测及分子对接筛选潜在靶点和通路。结果: 雷帕霉素成功诱导急性胸腺萎缩，模型对照组胸腺指数降低约50%（P<0.01），四肢抓力显著减弱（P<0.05）且未观察到明显毛发再生迹象。组织学显示胸腺皮质面积减少，皮髓质分界模糊，皮质TEC胞突网络连续性破坏，胸腺细胞发育早期受阻，空间分布紊乱，胸腺输出受抑制。与正常再生组比较，当归中、大剂量组能剂量依赖性增加胸腺皮质面积，显著提升皮质CK8+胸腺上皮细胞密度并修复其胞突网络连续性（P<0.01），同时上调Foxn1及其下游靶基因Dll4表达（均P<0.05）。在胸腺细胞发育层面，当归（大剂量）显著提升了CD3+TCRβ+胸腺细胞比例（P<0.05），促进CD4+CD8+细胞向功能成熟的单阳性胸腺细胞均衡分化，并重建了胸腺细胞正常空间定位，表现为皮质CD8+及CD4+CD8+细胞密度增加，TCRβ+细胞重新聚集于髓质区。此外，当归（中剂量）显著降低了胸腺促炎因子TNF-α、TGF-β及IGFBP5的mRNA水平（P<0.05），改善局部炎症微环境。在外周稳态方面，当归能维持并增加外周血CD3+T细胞比例，同时各剂量组均能增加CD34+造血干细胞的比例（P<0.05），并上调胸腺归巢因子Ccl25表达。网络药理学分析结果提示，当归活性成分可能靶向Wnt通路;实验验证发现，当归（中剂量）能显著上调胸腺内Wnt4 mRNA表达，抑制Gsk3β，并提高CTNNB1水平（P<0.05），表明其通过激活Wnt/CTNNB1/Foxn1信号通路驱动皮质TEC骨架修复与胸腺再生。结论: 当归通过激活Wnt/CTNNB1/Foxn1信号通路，并改善胸腺炎症微环境，协同促进皮质胸腺上皮细胞骨架修复与造血干细胞归巢，进而有效促进短期雷帕霉素诱导的急性胸腺退化后的皮质再生与功能恢复，可能具有抗免疫衰老的潜在益处。.
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