SD大鼠外阴慢性单纯性苔藓模型的构建及蛋白酶激活受体2的表达

Establishment of a SD rat model of vulvar lichen simplex chronicus and detection of the expression of protease activated receptor 2

目的建立SD大鼠外阴慢性单纯性苔藓(Lichen simplex chronicus,LSC)动物模型,研究蛋白酶激活受体2(protease activated receptor 2,PAR2)在模型大鼠外阴皮损中的表达,为LSC提供理想的动物模型。方法将70只雌性SD大鼠随机分为A组(空白对照组,n=10)、B组(丙酮对照组,n=10)和C组(单纯机械刺激对照组,n=10)、D组(实验组,n=40)。A组不予以任何处理;B组每周3次涂丙酮溶液于大鼠外阴皮肤,共10周;C组每周3次对大鼠皮肤予以单纯的机械刺激,共10周;D组予以0.5%二甲基苯并蒽(7,12-Dimethylbenzanthracene,DMBA)丙酮溶液联合机械刺激每周3次诱导大鼠外阴皮肤,共10周。6、8、10、12、14周肉眼及镜下观察大鼠外阴皮肤的变化。免疫组织化学、Western blotting检测A组和D组PAR2蛋白的表达,q RT-PCR检测PAR2的m RNA的表达。结果 D组第8周有57.5%(23/40)的SD大鼠出现了LSC改变,第10周有95%(38/40)的大鼠外阴表现为LSC,至12周时,20%(8/40)的大鼠外阴都形成了乳头状新生物。丙酮及单纯的机械刺激无导致LSC的作用。免疫组化及Western blotting结果显示D组PAR2的蛋白表达量较A组明显增加(P<0.05);q RT-PCR结果显示D组PAR2的m RNA的表达量较A组明显增加(P<0.05)。结论机械刺激联合0.5%DMBA丙酮溶液可诱导雌性SD大鼠LSC,PAR2可能与疾病的发生及进展密切相关。

Objective To establish a SD rat model of vulvar lichen simplex chronicus （LSC） and investigate the expression of protease activated receptor 2 （PAR2） in the genital skin. Methods Seventy female SD rats were randomly divided into group A （blank control group, n=10）, group B （with application of acetone solution 3 times per week for 10 weeks, n=10）, group C （with chronic mechanical irritation 3 times per week for 10 weeks, n=10）, and group D （with topical treatment with 0.5% 7, 12-Dimethylbenzanthracene [DMBA] in acetone solution and chronic mechanical irritation 3 times per week for 10 weeks, n=40）. The changes of the genital skin changes were observed regularly and the expression of PAR2 in groups A and D was detected with immunohistochemistry, Western blotting and qRT-PCR. Results In group D, LSC occurred in 23 rats （57.5%） at 8 weeks and in 38 rats （95%） at 10 weeks;8 rats （20%） showed papilloma at 12 weeks. Acetone treatment or chronic mechanical irritation did not cause LSC in the rats. Immunohistochemistry, Western blotting and qRT-PCR showed significantly increased expressions of PAR2 in group D at both the protein and mRNA levels as compared with those in group A （P〈0.05）. Conclusion 0.5%DMBA in acetone solution along with chronic mechanical irritation can induce LSC in female SD rats, and PAR2 is closely related with the occurrence and progression of LSC.