四氢姜黄素纳米乳凝胶的构建及其对特应性皮炎的作用与机制研究OA

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(10mg·kg-1 dexamethasone 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-1 Thc@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 high-dose 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(cGAS-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)mV,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,and 1.65±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.