补体C3调控细胞凋亡抑制咪喹莫特诱导小鼠银屑病样皮肤炎症反应  

Complement C3 Inhibits IMQ-Induced Psoriatic Skin Inflammation in Mice via Regulating Cell Apoptosis

作　　者：郑权友 杨奕 梁申菊 ZHENG Quanyou;YANG Yi;LIANG Shenju(Department of Nephrology and Urology,the 958th Hospital/the First Affiliated Hospital of Army Medical University,Chongqing 400020,China;Department of Rheumatism and Immunology,Army Special Medical Central,Army Medical University,Chongqing 400042,China)

机构地区：[1]陆军军医大学第一附属医院第九五八医院肾病泌尿科,重庆400020 [2]陆军军医大学陆军特色医学中心风湿免疫科,重庆400042

出　　处：《中国皮肤性病学杂志》2023年第3期256-264,298,共10页The Chinese Journal of Dermatovenereology

基　　金：国家自然科学基金项目(81900628);重庆市自然科学基金项目(cstc2018jcyjAX0260)。

摘　　要：目的利用咪喹莫特(IMQ)诱导小鼠银屑病样皮炎模型,探讨补体C3在该病理过程中的作用及机制。方法建立IMQ诱导银屑病样皮炎模型,比较C3^(+/+)及C3^(-/-)小鼠皮炎程度及病理改变;IHC检测皮损表皮细胞增殖,T细胞、中性粒细胞及巨噬细胞浸润,凋亡相关蛋白表达;TUNEL检测皮肤细胞凋亡;FCM比较小鼠皮损局部白细胞及中性粒细胞浸润及腋窝引流淋巴结IFN-γ应答。Z-VAD-FMK阻断凋亡途径,检测上述指标变化。结果C3^(-/-)小鼠皮炎较C3^(+/+)组明显加重,表现为:角质形成细胞增生、微脓肿、棘皮症及炎症细胞浸润等显著增加(HE),角质形成细胞异常增殖(Ki67)、T细胞(CD3和CD4)、中性粒细胞(Ly-6G)及巨噬细胞(F4/80)浸润明显增加(IHC),凋亡相关蛋白(Cleaved casp3、Cyt C及BAK)表达显著上调(IHC),TUNEL阳性凋亡细胞明显增加(IF)。此外,FCM发现C3^(-/-)小鼠皮损局部白细胞(CD45)及中性粒细胞(Gr-1)浸润较C3^(+/+)组显著增加;C3^(-/-)小鼠腋窝引流淋巴结IFN-γ^(+)CD3+T及IFN-γ^(+)CD8^(+)T频率明显增加。Z-VAD-FMK阻断凋亡途径后C3^(-/-)小鼠皮炎明显减轻,角质形成细胞增殖减弱,TUNEL阳性细胞减少,CD3+T细胞浸润减少,IFN-γ应答明显减弱。结论补体C3可能通过调节细胞凋亡抑制IMQ诱导银屑病样皮肤炎症反应。Objective To explore the effects and underlying mechanism of complement C3 in imiquimod(IMQ)-induced psoriatic skin inflammation model of mice.Methods IMQ-induced psoriatic skin inflammation model was established,and the severity of dermatitis and pathological changes in C3^(+/+)and C3^(-/-)mice were compared.Proliferation of epidermal cells,infiltration of T cells,neutrophils and macrophages,and expression of apoptosis-related proteins were detected by IHC staining,and the apoptotic cells in the skin were also detected by TUNEL staining.The infiltrated leukocytes and neutrophils in skin lesions and IFN-γresponses in draining lymph nodes were assessed by FCM.Before IMQ application,Z-VAD-FMK was used to block the apoptotic pathway and the changes of the above indicators were detected.Results Dermatitis in C3^(-/-)mice was significantly worse than that in C3^(+/+)group,which was manifested as significantly increased keratinocyte hyperplasia,microabscess,acanthosis and inflammatory cell infiltration by HE,the abnormal proliferation of keratinocytes(Ki67),significantly enhanced infiltration of T cells(CD3 and CD4),neutrophils(Ly-6G)and macrophages(F4/80)by IHC,significantly upregulated expression of apoptosis-related proteins(Cleaved casp3,Cyt C and BAK)by IHC,and significantly increased number of TUNEL positive apoptotic cells by IF.Moreover,FCM results indicated that the infiltration of leukocytes(CD45)and neutrophils(Gr-1)in the local lesions of C3^(-/-)mice was significantly increased compared with the C3^(+/+)group,and that the frequency of IFN-γ^(+)CD3+T and IFN-γ^(+)CD8^(+)T in draining lymph nodes of C3^(-/-)mice was also significantly increased.Blocking apoptosis with Z-VAD-FMK in C3^(-/-)mice clearly attenuated psoriatic skin inflammation,as proved by decreased keratinocyte proliferation,impaired TUNEL positive staining,attenuated CD3+T cells infiltration and down-regulated IFN-γresponse.Conclusion Complement C3 inhibits IMQ-induced psoriatic skin inflammation via regulating cell apoptosis.

关 键 词：银屑病 炎症反应 补体C3 细胞凋亡 

分 类 号：R758.63[医药卫生—皮肤病学与性病学]