摘要
目的 观察动力相关蛋白Drp1抑制剂Mdivi-1对咪喹莫特诱导的小鼠银屑病样皮炎的作用,并探讨其作用机制。方法 雌性8周龄C57BL/6小鼠随机分为对照组、咪喹莫特(IMQ)模型组、IMQ+Mdivi-1实验组。使用咪喹莫特构建小鼠银屑病样模型,实验组予以Mdivi-1腹腔注射,对照组及模型组予以等剂量溶剂腹腔注射。于造模第7天处死小鼠并取材。用PASI评分评判小鼠皮损严重程度,冰冻切片荧光染色检测小鼠皮肤组织中ROS含量,HE染色观察皮损组织形态学变化,免疫组织化学染色检测小鼠皮肤内Drp1蛋白的表达,Western blot检测小鼠皮肤组织中Drp1、NLRP3、IL-1β蛋白水平,ELISA技术检测小鼠血清中IL-17A、IL-18的表达。结果 模型组小鼠背部皮肤出现红斑、鳞屑、增厚等典型银屑病样皮损,实验组皮损明显减轻。实验组的PASI评分明显低于模型组。HE染色提示,实验组小鼠背部皮肤表皮厚度明显低于模型组,且Munro微脓肿明显减少。ROS荧光染色提示实验组ROS含量明显低于模型组。免疫组化结果显示,Drp1蛋白在实验组中表达明显低于模型组。Western blot结果显示,Drp1、NLRP3、IL-1β在实验组中的表达量明显低于模型组。ELISA结果提示,实验组小鼠血清中IL-17A、IL-18的含量低于模型组。结论 Mdivi-1可通过抑制动力相关蛋白Drp1的表达,减轻咪喹莫特诱导的小鼠银屑病样皮炎。
To investigate the effect of mitochondrial division inhibitor 1(Mdivi-1) on imiquimod(IMQ)-induced psoriasis-like skin inflammation in mice and its mechanism,female 8-week-old C57BL/6 mice were recruited and randomly divided into control group,IMQ model group,IMQ+Mdivi-1 experiment group.IMQ was used to induce the psoriasis-like skin inflammation model in mice.The mice in the experiment group were injected intraperitoneally(i.p.) with Mdivi-1,and the mice in the control group and model group were injected with the same volume of solvent.The mice were sacrificed on the 7th day for sampling.Psoriasis area and severity index(PASI)score was used to evaluate the severity of skin lesions in each group;the reactive oxygen species(ROS) content in skin tissue was detected by fluorescence staining of frozen section;HE staining was used to observe the histomorphologic change of skin lesions;immunohistochemical staining was used to detect the expression of dynamin-related protein 1(Drp1) in the skin of mice;Western blot was used to detect the protein levels of Drp1,NLRP3 and IL-1β in the skin tissues of mice in each group;and the expressions of IL-17A and IL-18 in mouse serum were detected by ELISA.Data showed that the model group had typical psoriatic lesions such as erythema,scale and thickening,and the Mdivi-1 group demonstrated obvious reduction of the lesions.The PASI score of the experiment group was significantly lower than that of the model group.HE staining indicated that the epidermal thickness of the back skin in the treatment group was significantly lower than that in the model group,and Munro microabscess was significantly reduced.ROS fluorescence staining indicated that ROS content in the experiment group was significantly lower than that in the model group;immunohistochemical results showed that the expression of Drp1 protein in the experiment group was significantly lower than that in the model group;Western blot results showed that the expression levels of Drp1,NLRP3 and IL-1β in the experiment group were significantly lower than those in the model group;ELISA results indicated that the expressions of IL-17A and IL-18 in serum of mice in the experiment group were lower than those in the model group.Taken together,Mdivi-1 can reduce mitochondrial damage and ROS production by inhibiting the expression of Drp1,thereby reducing the production of NLRP3 inflammasome,down-regulating IL-1β,IL-18 and IL-17A,and alleviating the IMQ-induced psoriasis-like skin inflammation in mice.
作者
顾玉洁
熊莉
吴倩
杨文翠
李元朝
周春丽
王儒鹏
GU Yujie;XIONG Li;WU Qian;YANG Wencui;LI Yuanchao;ZHOU Chunli;WANG Rupeng(Department of Dermatology and Rheumatology Immunology,Xinqiao Hospital,Army Medical University,Chongqing 400037,China;Department of Dermatology,General Hospital of Central Theater Command of PLA,Wuhan 430070,China)
出处
《免疫学杂志》
CAS
CSCD
2024年第1期59-64,共6页
Immunological Journal
