【摘要】 目的 构建四氢姜黄素纳米乳凝胶（Thc@NE@gel）的经皮递送系统，并对其在特应性皮炎（AD）中的作用效果和机制进行探讨。方法 用高能乳化联合低温溶胀法制备Thc@NE@gel，对制剂进行粒径、电位、载药量、包封率、形态及理化表征的测量，用Franz扩散池进行离体皮肤透皮实验。构建AD小鼠模型，将造模成功小鼠随机分为动物空白组、动物模型组、阳性药组(10 mg·kg^-1地塞米松凝胶)、Thc@NE组(20 mg·kg^-1Thc@NE)和动物Thc@NE@gel低、中、高剂量实验组(10、20、30 mg·kg^-1Thc@NE@gel)，每组6只;对各组小鼠称量并记录体质量变化;用苏木精伊红（HE）染色法观察皮肤组织病理损伤情况;用酶联免疫吸附试验（ELISA）法检测血清炎症因子水平;同时构建人永生化角质形成细胞（Ha Ca T）细胞体外炎症模型，将细胞分为细胞空白组、细胞模型组(LPS 1μg·mL^-1)、细胞Thc@NE@gel低、高剂量实验组（10、30μM Thc@NE@gel），对各组细胞用流式细胞仪检测氧化应激指标活性氧（ROS）;用酶联免疫吸附试验（ELISA）法检测细胞炎症因子水平;用蛋白质印迹法检测环鸟苷酸-腺苷酸合成酶-干扰素基因刺激蛋白信号（c GAS-STING）通路节点蛋白表达。结果 开发出了粒径为（195.30±5.2） nm，电位为（-24.51±1.45） m V，包封率为（72.70±0.67）%的Thc@NE@gel。动物Thc@NE@gel低、中、高剂量试验组、动物空白组、动物模型组、Thc@NE组和阳性药组的小鼠第17 d体质量分别为（21.50±1.20）、（22.30±1.15）、（24.00±1.25）、（26.20±1.30）、（17.30±1.10）、（18.20±1.15 ）和（17.50±1.05 ） g;免疫球蛋白E（IgE）水平分别为（40.52±10.23）、（32.84±9.65）、（30.67±10.12）、（21.23±3.45）、（53.21±4.12）、（56.45±5.89）和（64.32±12.14） ng·mL^-1;白细胞介素-17（IL-17）水平分别为（48.23±8.12）、（25.43±4.67）、（30.21±9.87）、（23.45±5.12）、（84.56±12.34）、（54.12±5.67）和（47.89±6.78） pg·mL^-1; IL-23水平分别为（45.67±3.12）、（44.23±2.98）、（40.12±2.45）、（25.34±7.12）、（73.45±15.67）、（55.23±6.78）和（24.56±1.89） pg·mL^-1;动物模型组与空白组比校，上述炎症指标在统计学上均有统计学意义（均P<0.05），与动物模型组相比，动物Thc@NE@gel中、高剂量组的IgE和IL-17指标，以及各剂量组的IL-23指标，在统计学上差异均有统计学意义（均P<0.05）;阳性药组的IgE水平与动物模型组比较，在统计学上差异无统计学意义（P > 0.05）。此外，细胞Thc@NE@gel低剂量、高剂量实验组、细胞空白组和细胞模型组的c GAS蛋白相对表达水平分别为1.15±0.12、0.82±0.08、1.00±0和1.65±0.18; STING相对蛋白表达量分别为0.95±0.10、0.60±0.06、1.00±0和1.65±0.15;细胞模型组的上述各项指标与细胞空白组比较，在统计学上差异均具有统计学意义（均P<0.001 ）。结论 Thc@NE@gel增强了Thc的抗炎效果，为天然活性成分的皮肤递送系统开发提供了新思路;此外，Thc@NE@gel通过抑制c GAS-STING改善特应性皮炎，为特应性皮炎的治疗提供新的机制思考。更多还原

【Abstract】 Objective To develop a transdermal delivery system for tetraphydrocurcumin nanoemulsion-gel（ Thc@NE@gel）,and to investigate its therapeutic efficacy and mechanism of action in atopic dermatitis（ AD）.Methods Thc@NE@gel was prepared using a high-energy emulsification method combined with a low-temperature swelling process.The formulation was characterized for particle size,zeta potential,drug loading,encapsulation efficiency,morphology and physicochemical properties.In vitro transdermal absorption experiments were conducted using a Franz diffusion cell.An AD mouse model was established,and successfully induced mice were randomly divided into animal control group,positive drug group( 10 mg·kg^-1dexamethasone gel),animal model group,Thc@NE group( Thc@ NE 20 mg·kg^-1) and animal Thc@NE@gel low-,medium-and high-dose experimental groups( 10,20,and 30 mg·kg^-1Thc@NE@gel),with 6 mice in each group.Body weights of mice in each group were measured and changes recorded; hematoxylin and eosin（ HE） staining was performed to observe histopathological damage in skin tissue; enzyme-linked immunosorbent assay（ ELISA） was used to detect serum inflammatory cytokine levels.Concurrently, an in vitro inflammatory model was established using human immortalized keratinocytes（ HaCaT）.Cells were divided into cell control group,cell model group( LPS 1 μg·mL^-1),cell low-and highdose Thc@NE@gel expreimental groups（ 10 and 30 μM Thc @ NE @ gel）.Reactive oxygen species（ ROS） were measured in each group using flow cytometry; serum inflammatory cytokine levels were detected by ELISA; Western blot analysis was performed to detect the expression of key proteins in the cyclic GMP-AMP synthase-stimulator of interferon genes（ c GAS-STING） signaling pathway.Results Thc@NE@gel was developed with a particle size of（ 195.30 ± 5.20） nm,a zeta potential of（-24.51 ± 1.45） m V,and a encapsulation efficiency of（ 72.70 ± 0.67） %.The body weights of mice in the animal Thc@NE@gel low-,medium-,and high-dose treatment groups,the animal blank group,the animal model group,the Thc@ NE group,and the positive control（ dexamethasone） group on day 17 were（ 21.50 ± 1.20）,（ 22.30 ± 1.15）,（ 24.00 ± 1.25）,（ 26.20 ± 1.30）,（ 17.30 ± 1.10）,（ 18.20 ± 1.15） and（ 17.50 ± 1.05） g,respectively; immunoglobulin E（ IgE） levels were（ 40.52 ± 10.23）,（ 32.84 ± 9.65）,（ 30.67 ± 10.12）,（ 21.23 ± 3.45）,（ 53.21 ± 4.12）,（ 56.45 ± 5.89） and（ 64.32 ± 12.14）ng·mL^-1; IL-17 levels were（ 48.23 ± 8.12）,（ 25.43 ± 4.67）,（ 30.21 ± 9.87）,（ 23.45 ± 5.12）,（ 84.56 ± 12.34）,（ 54.12 ± 5.67）,and（ 47.89 ± 6.78） pg · mL^-1; IL-23 levels were（ 45.67 ± 3.12）,（ 44.23 ± 2.98）,（ 40.12 ± 2.45）,（ 25.34 ± 7.12）,（ 73.45 ± 15.67）,（ 55.23 ± 6.78）,and（ 24.56 ± 1.89）pg·mL^-1; when comparing the animal animal model group with the animal blank group,all of the above inflammatory markers showed statistically significant differences（ all P < 0.05）.Compared with the animal model group,the IgE and IL-17 levels in the animal medium-and high-dose groups of Thc@NE@gel,as well as the IL-23 levels in all dose groups,showed statistically significant differences（ all P < 0.05）; the IgE levels in the positive control group showed no statistically significant difference compared with the animal model group（ P > 0.05）.Furthermore,the relative protein expression levels of cGAS in the low-dose and high-dose cell Thc@NE@gel experimental groups,the cell blank group,and the cell model group were 1.15 ± 0.12,0.82 ± 0.08,1.00 ± 0,and 1.65 ± 0.18,respectively; the relative protein expression levels of STING were 0.95 ± 0.10,0.60 ± 0.06,1.00 ± 0.00,and1.65 ± 0.15,respectively.Compared with the animal blank group,all of the aforementioned indicators in the cell model group showed statistically significant differences（ all P < 0.001）.Conclusion Thc@NE@gel enhances the anti-inflammatory effects of Thc,providing new insights for the development of skin delivery systems for natural active ingredients; furthermore,by inhibiting the cGAS-STING pathway,Thc@NE@gel improves atopic dermatitis,offering a new mechanistic perspective for its treatment.更多还原