目的 基于瞬时受体电位A1（TRPA1）设计开发一种具有改善特应性皮炎（AD）作用的天然活性成分乳膏剂。方法 构建TRPA1通道高表达细胞，采用细胞荧光钙离子成像技术从31种活性天然产物中筛选TRPA1抑制剂，并筛选最佳组合以及比例；通过单因素实验和响应面分析优化乳膏剂中甘油、聚丙烯酸钠（SP）、辛酸/癸酸甘油三脂（M318）的添加量。构建大鼠皮肤紫外线B（UVB）晒伤模型，通过检测皮肤机械痛阈评价乳膏剂对痛觉的缓解作用。通过2，4-二硝基氟苯（DNFB）诱导构建小鼠AD模型，评价乳膏剂对瘙痒、皮损评分、皮肤病理损伤以及皮肤中白细胞介素（IL）-6、IL-4、IL-33、IL-1β、γ干扰素（IFN-γ）、肿瘤坏死因子α（TNF-α）炎症因子mRNA的表达水平的改善效果。结果 筛选出对TRPA1具有抑制作用的9种活性天然产物，发现白藜芦醇与和厚朴酚物质的量比值为5∶1时协同效果最佳，确定为白藜芦醇与和厚朴酚双因组合（BHSY）。经配方优化BHSY乳膏剂的最佳组成为：甘油5.13%、SP 0.91%、M318 3.01%，其中BHSY质量分数为0.17%～0.50%，其粒径分布均匀且稳定性好。与模型组比较，BHSY乳膏剂在UVB照射损伤后的第2、3天可剂量相关性地提高大鼠皮肤痛阈（P＜0.05、0.01、0.001）；并可明显降低AD小鼠的搔抓次数（P＜0.05、0.01），改善皮损评分（P＜0.05、0.01），抑制背部表皮增厚（P＜0.001）及皮肤中炎症因子IL-6、IL-4、IL-33、IL-1β、INF-γ、TNF-α mRNA的表达（P＜0.05、0.01、0.001）。结论 基于TRPA1通道开发了具有协同增效作用的活性天然产物BHSY乳膏剂，对减轻皮肤的UVB损伤以及舒缓AD症状具有显著效果。

Objective Based on transient receptor potential A1 (TRPA1), a natural active ingredient cream preparation with the effect of improving atopic dermatitis (AD) was developed and optimized. Methods TRPA1 overexpressed cells were constructed to be used in the screening of TRPA1 inhibitors from 31 natural compounds by cell fluorescence calcium ion imaging, and the optimal combination and proportion were screened. The addition amounts of glycerol, sodium polyacrylate (SP), and glyceryl tristearate (M318) in the cream were optimized through single-factor experiments and response surface analysis. A rat skin UVB sunburn model was established to evaluate the analgesic effect of the cream by detecting the mechanical pain threshold of the skin. A mouse AD model was induced by 2, 4-dinitrofluorobenzene (DNFB) to assess the improvement effect of the cream on pruritus, skin lesion score, skin pathological damage, and the mRNA expression levels of inflammatory factors such as interleukin (IL)-6, IL-4, IL-33, IL-1β, interferon-γ (IFN-γ), and tumor necrosis factor α (TNF-α) in the skin. Results Nine active natural products with inhibitory effects on TRPA1 were screened out. It was found that the best synergistic effect was achieved when the molar ratio of resveratrol to magnolol was 5∶1, and this combination was determined as the resveratrol and magnolol dual-component combination (BHSY). After formula optimization, the optimal composition of the BHSY cream was: glycerol 5.13%, SP 0.91%, M318 3.01%, with BHSY content ranging from 0.17% to 0.50%. The particle size distribution was uniform and the stability was good. Compared with the model group, the BHSY cream could dose-dependently increase the mechanical pain threshold of rat skin on the 2nd and 3rd days after UVB irradiation injury (P < 0.05, 0.01, 0.001); it could also significantly reduce the scratching frequency of AD mice (P < 0.05, 0.01), improve the skin lesion score (P < 0.05, 0.01), inhibit the thickening of the epidermis on the back (P < 0.001), and reduce the mRNA expression of inflammatory factors such as IL-6, IL-4, IL-33, IL-1β, INF-γ, and TNF-α in the skin (P < 0.05, 0.01, 0.001). Conclusion Based on the TRPA1 channel, an active natural product BHSY cream with synergistic effects has been developed, which demonstrates significant efficacy in alleviating UVB-induced skin damage and mitigating AD symptoms.

R285.5

广西科技厅“带土移植”人才引育计划(AA23026008)