青鹏软膏对皮肤干燥志愿者及特应性皮炎样小鼠模型皮肤屏障功能的影响

Effects of Qingpeng ointment on skin barrier function in volunteers with xerosis and atopic dermatitis-like mouse models

目的 探讨青鹏软膏对皮肤屏障功能的影响及其可能机制。 方法 招募12名小腿伸侧皮肤干燥的女性志愿者，青鹏软膏涂于右小腿伸侧（青鹏侧）、青鹏软膏基质涂于左小腿伸侧（基质侧），用药7 d。分别于用药前、用药3 d及用药7 d测定皮肤屏障功能相关指标，如，经表皮失水量（TEWL）及角质层含水量。2,4-二硝基氟苯诱导小鼠背部产生特应性皮炎样改变，分别外用青鹏软膏基质（基质组），50%、75%、100%青鹏软膏治疗（50%、75%、100%青鹏组），每天2次，用药2周后，进行组织病理检查，背部皮褶厚度、TEWL的测定，免疫组化测定表皮丝聚蛋白，外皮蛋白及激肽释放酶7含量。 结果 志愿者外用青鹏软膏及其基质，3 d、7 d后均较治疗前TEWL显著下降（青鹏侧t = 2.651、3.615；基质侧t = 2.996、3.586，均P 〈 0.05），含水量显著上升（青鹏侧t = 9.029、13.842；基质侧t = 5.830、11.299，均P 〈 0.001）。青鹏侧及基质侧TEWL、含水量比较，差异无统计学意义（P 〉 0.05）。从外观、皮褶厚度及组织病理看，基质组小鼠与模型组相比，皮炎程度差异不大，各青鹏组小鼠皮炎均有不同程度缓解。基质组、青鹏组TEWL均较模型组下降（P 〈 0.05），而基质组与青鹏组TEWL比较，差异无统计学意义（P 〉 0.05）。基质组、青鹏组KLK7表达均较模型组下降（P 〈 0.05），但各治疗组间比较，差异无统计学意义（P 〉 0.05）。 结论 青鹏软膏基质有改善皮肤屏障功能的作用。

Objective To estimate effects of Qingpeng ointment on skin barrier function, and to explore their potential mechanisms. Methods A total of 12 female volunteers with xerosis on the extensor surfaces of legs were enrolled into this study. All the volunteers were topically treated with Qingpeng ointment on the right leg （Qingpeng side） and its vehicle on the left leg （vehicle side） twice daily for 7 consecutive days. Skin barrier function-associated indexes including transepidermal water loss （TEWL） and the water content of the stratum corneum were measured before the treatment, after 3- and 7-day treatment separately. Thirty-six BALB/c female mice were randomly and equally divided into 6 groups： a blank control group receiving no induction or treatment, a model group induced by 2, 4-dinitrofluorobenzene （DNFB） on the back for 5 consecutive weeks, 3 Qingpeng groups and a vehicle group induced by DNFB on the back for 5 consecutive weeks followed by topical treatment with different concentrations （50%, 75%, 100%） of Qingpeng ointment or its vehicle twice a day for 2 consecutive weeks. At the end of treatment, skin appearance of mice was observed with naked eyes, skinfold thickness and TEWL were measured. Then, all the mice were sacrificed, and skin tissue specimens were resected from the back of mice followed by histopathologic examination and immunohistochemical staining for the detection of filaggrin （FLG）, involucrin （IVL） and kallikrein 7 （KLK7） expressions in the epidermis. Results After start of treatment, both the Qingpeng side and vehicle side showed significantly decreased TEWL on day 3 （t = 2.651, 2.996, respectively, both P 〈 0.05） and 7 （t = 3.615, 3.586, respectively, both P 〈 0.05）, but increased water content of the stratum corneum on day 3 （t = 9.029, 5.830, respectively, both P 〈 0.001） and 7 （t = 13.842, 11.299, respectively, both P 〈 0.001） compared with those before the treatment. However, no significant differences were observed in TEWL or the water content of the stratum corneum between the Qingpeng side and vehicle side at any of the time points （all P 〉 0.05）. In addition, there were no marked differences in mouse skin appearance, skinfold thickness or histopathologic manifestations between the vehicle group and model group, while the manifestations of dermatitis were attenuated to different extents in the three Qingpeng groups compared with the model group. Both TEWL and KLK7 expression levels were significantly decreased in the vehicle group and three Qingpeng groups compared with the model group （all P 〈 0.05）, but similar between the vehicle and Qingpeng groups （all P 〉 0.05）. Conclusion The vehicle of Qingpeng ointment can improve skin barrier function.