青刺果油对特应性皮炎样小鼠模型表皮通透屏障及抗菌肽表达的影响

The Study of Prinsepia Utilis Royle Oil Effects on Epidermal Barrier and Antimicrobial Peptides in Atopic Dermatitis-like Mouse Model

目的探讨青刺果油在修复表皮通透屏障功能的同时对表皮抗菌肽表达的影响。方法将BALB/c小鼠随机分为6组:正常(A)组8只、正常+2%青刺果油(B)组6只、模型(C)组8只、模型+2%青刺果油(D)组6只、模型+基质(E)组6只、模型+2%橄榄油(F)组6只。利用1-氯-2,4-二硝基苯(DNCB)和4-乙氧基亚甲基-2-苯基-2-唑啉-5-酮(OX)于C、D、E、F组小鼠背部构建特应性皮炎样模型,然后在B、D组小鼠背部涂抹2%青刺果油,在E组小鼠背部涂抹基质(辛酸/癸酸甘油三酯),在F组小鼠背部涂抹2%橄榄油。分别于干预前、干预3、7、15天用无创性皮肤生理功能测试仪测定小鼠背部受试区经表皮水分流失(TEWL)及皮肤水分,15天后取小鼠背部受试区皮损,用Western blot方法测定cathelicidin和β-防御素(MBD1、2、3、4)的表达。结果无创性皮肤生理功能测试结果显示,用2%青刺果油干预后,D组TEWL值均明显下降,皮肤水分均明显上升;与C组相比,D组TEWL值干预3、15天后明显下降;与E组相比,D组TEWL值干预3、7、15天后均明显下降;D组皮肤水分干预7、15天后明显上升;与F组相比,D组TEWL值干预7、15天后明显下降,D组皮肤水分干预7、15天后明显上升,差异均有统计学意义(P均<0.05)。Western blot结果显示,用2%青刺果油干预后,D组cathelicidin和MBD3蛋白的表达明显高于A、B、C、E、F组,D组MBD1、2、4蛋白的表达高于C、E、F组。结论青刺果油在修复表皮通透屏障功能的同时可通过增加cathelicidin和β-防御素的表达来增加表皮抗菌肽的表达,从而可用于AD的辅助治疗。

Objective To investigate the effect of prinsepia utilis royle oil（PURO）on the expression of epidermal antimicrobial peptides and reparation of epidermal barrier function. Methods BALB/c mice were randomly divided into 6 groups according to different treatments：normal group（A）8 mice,normal＋ 2% PURO（B）6 mice,model group（C）8 mice,model ＋ 2% PURO（D）6 mice,model ＋ matrix（E）6 mice,model ＋ 2% olive oil（F）6 mice. 1-chloro-2,4-dinitrobenzene（DNCB）and 4-ethoxymethylene-2-phenyl-2-oxazolin-5-one（OX）were used in modeling atopic dermatitis on the back skin of the mice in the group C, D, E and F. Then,2% PURO were applied in group B and D,and the matrix（octanoic acid / capric triglyceride）and 2% olive oil were coated on the back of the mice in group E and F,respectively. The time before the intervention,and 3,7 and 15 days after the intervention,transepidermal water loss（TEWL）and hydration of the test area on the back of the mice were measured with noninvasive methods. The expression of cathelicidin and β-defensins（MBD1,2,3,4）were evaluated by Western blot after 15 days. Results After eluting with 2% PURO intervention,TEWL of the group D decreased significantly and the hydration significantly increased. Compared with group C,the TEWL of group D was significantly lower after 3 and 15 days. The TEWL of the group E was higher after 3,7 and 15 days than the group D,and simultaneously the hydration of group E was much lower after 7 and 15 days. In the comparison with the group F,the TEWL of group D was lower than the group F after 7 and 15 days,and the hydration of group D was significantly higher after 7 and 15 days（all P〈0.05）. Western blot showed the expression of cathelicidin and MBD3 in group D was significantly higher than that in group A,B,C,E and F,and the expression of MBD1,2,4 in group D was higher than those in group C,E and F. Conclusion PURO can increase the expression of epidermal antimicrobial peptides through increasing the expression of cathelicidin and β-defensinsand promoterepairation of epidermal barrier function,which showed the protential of PURO in AD adjuvant therapy.