摘要
目的:探讨皮肤中半乳糖凝集素-3(Gal3)对银屑病小鼠模型中皮损处肥大细胞激活及银屑病皮炎损伤的影响及其潜在机制。方法:每日分别对SPF级Gal3野生型(Gal3^(+/+))和Gal3基因敲除(Gal3^(−/−))小鼠背部裸露皮肤处涂抹相同剂量咪喹莫特乳膏,连续5 d以构建银屑病小鼠模型。观察小鼠背部大体病变,并根据银屑病皮损面积和严重程度指数(PASI)进行评分,利用实时荧光定量PCR测定小鼠皮损组织细胞因子il-1、il-17表达,比较Gal3^(+/+)小鼠和Gal3^(−/−)小鼠银屑病炎症的严重程度;HE染色观察比较皮损组织病理学变化;甲苯胺蓝染色观察小鼠皮损组织中肥大细胞的形态和分布差异;免疫组织化学和图像分析技术检测肥大细胞活化标志物如类胰蛋白酶和5-羟色胺(5-HT)的表达、分布等差异。结果:连续应用咪喹莫特乳膏后,Gal3^(−/−)小鼠背部出现鳞屑、浸润斑块和红斑等典型银屑病临床表型,而Gal3^(+/+)小鼠背部出现细小鳞屑与少量浸润,Gal3^(−/−)小鼠PASI评分较Gal3^(+/+)小鼠明显升高;Gal3^(+/+)小鼠与Gal3^(−/−)小鼠皮损处il-1、il-17表达升高,但后者炎症因子表达上调更显著。组织病理学观察发现Gal3^(+/+)小鼠表皮稍增厚,Gal3^(−/−)小鼠表皮明显增厚,且网脊向下延伸部分伴有炎症细胞聚集;甲苯胺蓝染色表明Gal3^(+/+)小鼠皮损组织肥大细胞分布稀疏且大部分结构完整,而Gal3^(−/−)小鼠肥大细胞数量增多并广泛分布于皮肤组织,且多数肥大细胞处于活化脱颗粒状态;免疫组织化学染色观察发现Gal3^(−/−)小鼠皮损组织类胰蛋白酶和5-HT较Gal3^(+/+)小鼠表达信号明显增强,且多集中在表皮损伤处。结论:Gal3的缺失引起银屑病小鼠模型皮损处肥大细胞过度活化,加重银屑病皮肤炎症损伤发生和疾病进展,Gal3在该模型中对皮损处肥大细胞活化与脱颗粒有抑制作用。
Objective:To investigate the effects and potential mechanisms of galectin-3(Gal3)on mast cells activation and skin injury in a psoriatic murine model.Methods:Imiquimod was applied to the bare skin on back of SPF(Gal3^(+/+))mice and their matched(Gal3^(−/−))siblings respectively once daily for 5 d to establish a psoriatic mice model.The development and dynamics of skin lesions were monitored and recorded,scored.with the psoriasis area and severity index(PASI).The expressions of il-1 and il-17 in the damaged skin were detected by real-time PCR to compare the severity of inflammation between Gal3^(+/+)mice and their Gal3-/-siblings.HE staining was used to observe the histopathological changes of skin.The difference in morphology and distribution of mast cells were examined by toluidine blue staining.Immunohistochemistry and image analysis techniques were improved to assay the expression and distribution of activated mast cells markers such as tryptase and 5-hydroxytryptamine(5-HT).Results:The typical psoriatic signs including erythema,scaling and infiltration were formed in mice after continuous administration of imiquimod in Gal3^(−/−)mice,whereas Gal3^(+/+)mice just developed small scaling and mild infiltration.PASI score of Gal3^(−/−)mice was significantly higher than that of Gal3^(+/+)mice.In accordance,il-1 and il-17 were increased in the skin lesions of Gal3^(+/+)mice and Gal3^(−/−)mice,but these inflammatory factors were more upregulated remarkably in the latter.Histopathology observation revealed that the epidermis of Gal3^(+/+)mice was slightly thickened,whereas thickened epidermis of Gal3^(−/−)mice was more seriously and the rete ridges extended downward,with massive inflammatory cells aggregation.Toluidine blue staining indicated that the mast cells were sparsely distributed and most of their structures were intact in Gal3^(+/+)mice,instead the mast cells of Gal3^(−/−)mice were mostly in degranulation state with increased and distributed widely in skin.Immunohistochemistry staining revealed that tryptase and 5-HT,compared with those of Gal3^(+/+)mice,were increased obviously in the lesioned skin of Gal3^(−/−)mice,most of them were concentrated in the epidermal in particular.Conclusion:The deficiency of Gal3 may result in over-activation and degranulation of mast cells at the skin in psoriatic mice model,which aggravates the occurrence of dermatitis inflammatory injury and disease progression in psoriasis,and Gal3 plays suppressive effects on mast cells activation and degranulation at the skin lesions in this model.
作者
程美琦
金柱
张丁
王琼
胡悦
晁金
王德成
韩珊珊
CHENG Meiqi;JIN Zhu;ZHANG Ding;WANG Qiong;HU Yue;CHAO Jin;WANG Decheng;HAN Shanshan(Hubei Key Laboratory of Tumor Microenvironment and Immunotherapy(China Three Gorges University),China Three Gorges University Institute of Infection and Inflammation,Yichang 443002,China;The Third Peo-ple's Hospital of Yichang,Yichang 443003,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2024年第4期686-692,共7页
Chinese Journal of Immunology
基金
国家自然科学基金(31772709,82000914)。
