摘要
目的探讨γ分泌酶抑制剂阻断Notch-Hes1信号通路对银屑病样皮炎小鼠γδT17细胞表达的影响.方法将45只健康雄性SPF级BALB/c小鼠按照简单随机抽样法等分为对照组、模型组和干预组,模型组和干预组小鼠背部脱毛区均每日外涂5%咪喹莫特乳膏62.5 mg,干预组还在外用咪喹莫特后立即接受γ分泌酶抑制剂(DAPT)10 mg/kg腹腔注射,每日1次;对照组仅每日外涂等剂量白凡士林.共处理6d,采用银屑病皮损严重程度指数(PASI)评估皮损变化.第7天麻醉小鼠心脏取血,取脾脏及皮肤组织,制备单细胞悬液,比较计算各组小鼠脾指数,苏木精-伊红染色观察皮肤组织病理学改变.流式细胞仪检测脾脏及皮肤组织中γδT17细胞比例,实时定量反转录PCR检测脾细胞悬液中Hes1 mRNA表达水平,酶联免疫吸附实验检测血清中白细胞介素17A(IL-17A)含量.采用单因素方差分析和重复测量的方差分析比较各组间指标差异,Pearson相关分析评价不同指标间的相关性.结果末次处理24h后,肉眼观察显示,干预组小鼠银屑病样皮炎改变明显轻于模型组,PASI显著低于模型组,且表皮增厚及真皮炎细胞浸润程度明显轻于模型组.模型组小鼠脾指数(12.534±1.636)、脾脏(24.659%±4.603%)及皮肤组织γδT17细胞比例(22.127%±5.670%)、脾脏Hes1 mRNA表达水平(4.867±0.543)、血清IL-17A含量[(22.478±2.776)ng/L]均明显高于对照组(P<0.01);干预组上述指标分别为9.449±1.040、14.966%±5.770%、13.631%±5.946%、2.541±0.347、(18.639±1.816)ng/L,均显著低于模型组(P<0.01).模型组及干预组小鼠脾脏γδT17细胞比例与血清IL-17A含量均呈正相关(r值分别为0.56和0.53,均P<0.05),皮损γδT17细胞比例与PASI均呈正相关(r值分别为0.56和0.52,均P<0.05),脾脏Hes1 mRNA表达水平与γδT17细胞比例(r值分别为0.61和0.58)、血清IL-17A含量(r值分别为0.60和0.54)均呈正相关(P<0.05).结论γ分泌酶抑制剂阻断Notch-Hes1信号通路
Objective To evaluate the effect of Notch-Hes1 signaling blockade by a γ-secretase inhibitor on the expression of γδT17 cells in a mouse model of psoriasis-like skin inflammation.Methods Forty-five healthy specific pathogen-free(SPF)male BALB/c mice were randomly divided into control group,model group and intervention group by simple random sampling.The model group and intervention group were both topically treated with imiquimod 5%cream(62.5 mg once a day)on the shaved back,the intervention group were then intraperitoneally injected with the γ-secretase inhibitor DAPT(10 mg/kg once a day)immediately after topical application of imiquimod,and the control group were topically treated with equivalent amount of vaseline once a day.After 6-day treatment,psoriasis area and severity index(PASI)was used to evaluate changes of skin lesions.On day 7,blood samples were obtained from all the mice through heart puncture after anesthetization,and spleen and skin tissues were acquired to prepare single cell suspension.Spleen index was compared among the 3 groups.Skin tissues on the mouse back were resected and subjected to hematoxylin-eosin staining to observe histopathological changes.Flow cytometry was performed to determine the percentage of γδT17 cells in the spleen and skin tissues,real-time reverse transcription(RT)-PCR to measure the mRNA expression of Hes1 in single cell suspension of the spleen,and enzyme-linked immunosorbent assay(ELISA)to determine the serum level of interleukin(IL)-17A.Statistical analysis was carried out by using one-way analysis of variance and repeated measures analysis of variance for comparison of indices among groups,and Pearson correlation analysis for evaluating the correlation between different indices.Results Twenty-four hours after the final treatment,the intervention group showed milder psoriasis-like skin inflammation,lower PASI score,and milder degree of epidermal thickening and dermal inflammatory cell infiltration compared with the model group.The model group showed significan
作者
王燕芹
李新新
薛海波
纪洪
孙大康
马蕾
Wang Yanqin;Li Xinxin;Xue Haibo;Ji Hong;Sun Dakang;Ma Lei(Department of Dermatology,Binzhou Medical University Hospital,Binzhou 256603,Shandong,China;Department of Endocrinology and Metabolism,Binzhou Medical University Hospital,Binzhou 256603,Shandong,China;Department of Pathology,Binzhou Medical University Hospital,Binzhou 256603,Shandong,China;Department of Clinical Medical Laboratory,Binzhou Medical University Hospital,Binzhou 256603,Shandong,China)
出处
《中华皮肤科杂志》
CAS
CSCD
北大核心
2019年第11期801-807,共7页
Chinese Journal of Dermatology
基金
国家自然科学基金(81803145)
山东省医药卫生科技发展计划项目(2016WS0045)。
