目的 探讨阿胶在延缓D-半乳糖诱导的皮肤衰老及维持微血管稳态方面的药效作用，并阐明其通过酪氨酸激酶受体2（tyrosine kinase with immunoglobulin-like and EGF-like domains 2，Tie2）/血管生成素（angiopoietin，Ang）/血管内皮钙黏蛋白（vascular endothelial cadherin，VE-cadherin）信号轴调控血管成熟度的分子机制。方法 采用sc D-半乳糖构建小鼠早衰模型，连续8周ig给予阿胶（270.3 mg/kg）干预，通过激光散斑血流成像（laser speckle contrast imaging，LSCI）监测背部血流，光相干断层扫描血管造影（optical coherence tomography angiography，OCTA）观察微血管密度，苏木素-伊红（hematoxylin-eosin staining，HE）及Masson染色评估皮肤组织形态及胶原含量，免疫荧光及免疫组化检测皮肤组织血管内皮标志物CD31和周细胞标志物血小板源生长因受体β（platelet-derived growth factor receptor β，PDGFRβ）的表达，采用ELISA检测血清Ang1/Ang2的值，采用qRT-PCR检测皮肤组织白细胞介素-1β（interleukin-1β，IL-1β）、趋化因子配体1（C-X-C motif chemokine ligand 1，CXCL1）和肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）mRNA表达。采用过氧化氢（hydrogen peroxide，H₂O₂）诱导人脐静脉内皮细胞（human umbilical vein endothelial cells，HUVEC）构建应激性早衰模型，给予62.5～250.0 μg/mL阿胶预处理24 h，通过CCK-8检测细胞活力，通过衰老相关β-半乳糖苷酶（senescence-associated β-galactosidase，SA-β-gal）染色及活性氧（reactive oxygen species，ROS）水平评估衰老程度，qRT-PCR检测衰老相关分泌表型（senescence-associated secretory phenotype，SASP）及炎症相关基因表达，Western blotting检测肿瘤蛋白p53（tumor protein p53，p53）、周期蛋白依赖性激酶抑制剂1A（cyclin dependent kinase inhibitor 1A，p21）、p-Tie2、Tie2、VE-cadherin、血管内皮生长因子A（vascular endothelial growth factor A，VEGFA）蛋白表达，ELISA检测Ang1/Ang2的值，通过周细胞招募及内皮渗透实验评价血管稳定性。结果 动物实验中，阿胶显著改善衰老小鼠皮肤褶皱，增加表皮及真皮厚度和胶原容积分数（P＜0.01、0.001），恢复皮肤血流灌注并减少杂乱的无序血管生成，提高皮肤组织PDGFRβ的表达及血清Ang1/Ang2的值（P＜0.001），下调炎症因子表达（P＜0.001），增强血管成熟度。细胞实验中，阿胶显著降低HUVEC中SA-β-gal阳性率及ROS水平（P＜0.001），下调SASP及炎症相关基因表达（P＜0.01、0.001），上调p-Tie2和VE-cadherin的蛋白表达（P＜0.001），下调p53、p21、Tie2和VEGFA的蛋白表达（P＜0.05、0.001），并提高Ang1/Ang2的值（P＜0.05、0.01），显著增强内皮细胞对周细胞的招募能力及血管内皮屏障的完整性。结论 阿胶可改善D-半乳糖诱导的衰老皮肤模型的衰老表型，提高微血管成熟度并维持血管稳态，其作用可能与调节Tie2/Ang/VE-cadherin信号轴有关。

Objective To explore the pharmacological effect of Ejiao (Colla Corii Asini, CCA) on delaying D-galactose-induced skin aging and maintaining microvascular homeostasis, and to clarify the molecular mechanism of its regulation of vascular maturity through tyrosine kinase with immunoglobulin-like and EGF-like domains 2 (Tie2)/angiopoietin (Ang)/vascular endothelial cadherin (VE-cadherin) signal axis. Methods The model of premature senility in mice was established by sc D-galactose, and CCA (270.3 mg/kg) was administered ig for eight weeks. The back blood flow was monitored by laser speckle contrast imaging (LSCI), the microvessel density was observed by optical coherence tomography angiography (OCTA), the skin tissue morphology and collagen content were evaluated by hematoxylin-eosin staining (HE) and Masson staining, the expressions of vascular endothelial marker CD31 and pericyte marker platelet-derived growth factor receptor β (PDGFRβ) were detected by immunofluorescence and immunohistochemistry, the Ang1/Ang2 value in serum was detected by ELISA, the expressions of interleukin-1β (IL-1β), C-X-C motif chemokine ligand 1 (CXCL1) and tumor necrosis factor-α (TNF-α) mRNA in skin tissue was detected by qRT-PCR. Human umbilical vein endothelial cells (HUVEC) were induced by hydrogen peroxide (H₂O₂) to construct a stress premature aging model, cells were pretreated with 62.5—250.0 μg/mL CCA for 24 h. Cell viability was detected by CCK-8. Senescence-associated β-galactosidase (SA-β-gal) staining and reactive oxygen species (ROS) level were used to assess the degree of aging. qRT-PCR was used to detect the senescence-associated secretory phenotype (SASP) and inflammation related gene expressions. Western blotting was used to detect tumor protein p53 (p53), cyclin dependent kinase inhibitor 1 A (p21), p-Tie2, Tie2, VE-cadherin and vascular endothelial growth factor A (VEGFA) protein expressions. Ang1/Ang2 value was detected by ELISA. Vascular stability was evaluated through pericyte recruitment and endothelial permeability experiments. Results In the animal experiment, CCA significantly improved the skin fold of aging mice, increased the thickness of epidermis and dermis and collagen volume fraction (P < 0.01, 0.001), restored skin blood flow perfusion and reduced disordered angiogenesis, increased the expression of PDGFRβ in skin tissue and the value of Ang1/Ang2 in serum (P < 0.001), decreased the expressions of inflammatory factors (P < 0.001), and enhanced vascular maturity. In the cell experiment, CCA significantly reduced the positive rate of SA-β-gal and ROS level in HUVEC (P < 0.001), down-regulated the expressions of SASP and inflammation related genes (P < 0.01, 0.001), up-regulated the protein expressions of p-Tie2 and VE-cadherin (P < 0.001), down-regulated the protein expressions of p53, p21, Tie2 and VEGFA (P < 0.05, 0.001), and increased the value of Ang1/Ang2 (P < 0.05, 0.01), significantly enhancing the ability of endothelial cells to recruit pericytes and the integrity of vascular endothelial barrier. Conclusion CCA can improve the aging phenotype of aging skin model induced by D-galactose, improve microvascular maturity and maintain vascular homeostasis, which may be related to the regulation of Tie2/Ang/VE-cadherin signal axis.

R285.5

山东东阿阿胶股份有限公司研究基金资助项目（HX2023001）；中国中医科学院科技创新工程创新能力提升专项（NLTS2025004）；中央级公益性科研院所基本科研业务费专项（JJPY2025002）；泰山工业专项（tscy20180234）；中央引导地方科技发展资金（YDZX2023090）