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目的探讨磷脂酶Cε(PLCε)在日本血吸虫(SJ)尾蚴性皮肤炎症中的作用机制。方法用日本血吸虫尾蚴感染PLCε基因敲除(KO)和PLCε野生型(WT)两组小鼠,建立Ⅰ型和Ⅳ型超敏反应模型。于感染后不同时间点提取小鼠感染处皮肤组织,部分制作冰冻组织切片,用于HE染色检查和免疫荧光染色检查;部分组织提取总RNA,采用qRTPCR方法检测相关炎症细胞因子mRNA表达水平。结果 HE染色显示,Ⅰ型超敏反应模型中PLCεKO和WT两组皮肤肥厚度和炎性细胞浸润数量差异无统计学意义(P均>0.05)。在Ⅳ型超敏反应模型,KO小鼠炎症反应显著减弱,与WT组相比皮肤增厚程度减轻(t_(48h)=5.317,P=0.000t_(72h)=10.162,P=0.000),且炎性细胞浸润数量显著减少(t_(48h)=2.888,P=0.020)。qRT-PCR检测T细胞衍生因子IL-4,IFN,IL-17及IL-23mRNA表达在WT与KO组间差异无统计学意义(IL-4:t_(24h)=0.496,P=0.625;IFN:t_(24h)=-0.035,P=0.973;IL-17:t_(24h)=0.112,P=0.938;IL-23:t_(24h)=-1.151,P=0.261)。但成纤维细胞和角质形成细胞等体细胞所诱导的炎症细胞因子IL-1α,IL-1β以及趋化因子Cxcl-1,Cxcl-2mRNA表达在KO组受抑制(IL-1α:t_(12h)=4.785,P=0.000;t_(24h)=2.109,P=0.046;IL-1β t_(6h)=3.187,P=0.004;t_(12h)=5.049,P=0.000;Cxcl-1:t_(12h)=4.858,P=0.000;Cxcl-2:t_(24h)=7.957,P=0.000)。结论 PLCε在日本血吸虫尾蚴性皮炎中通过调控体细胞的细胞因子的表达而影响炎症反应。
Objective To investigate the mechanism of phospholipase Cε (PLCε) in the skin inflammation of Schistosoma japonicum (SJ). Methods Type I and type IV hypersensitivity models were established in mice infected with PLCε gene knockout (KO) and PLCε wild type (WT) with Schistosoma japonicum cercariae. At different time points after infection, skin tissues from mice were extracted and frozen tissue sections were partially prepared for HE staining and immunofluorescence staining. Total RNA was extracted from some tissues and qRTPCR method was used to detect the expression of related inflammatory cytokines mRNA. Results The results of HE staining showed that there was no significant difference in the number of skin hypertrophy and inflammatory cell infiltration between PLCεKO and WT groups in type Ⅰ hypersensitivity (P> 0.05). The inflammatory response in KO mice was significantly attenuated in type IV hypersensitivity model, and the skin thickening was relieved compared with WT group (t 48h = 5.317, P = 0.000t 72h = 10.162, P 0.000) The number of inflammatory cell infiltration was significantly reduced (t_ (48h) = 2.888, P = 0.020). There were no significant differences in the expression of IL-4, IFN, IL-17 and IL-23 mRNA between the WT and KO groups by qRT-PCR (IL-4: t_ (24h) = 0.496, P = : t_ (24h) = -0.035, P = 0.973; IL-17: t_ (24h) = 0.112, P = 0.938; IL-23: t_ (24h) = - 1.151, P = 0.261). However, the expressions of IL-1α, IL-1β, Cxcl-1 and Cxcl-2 mRNA induced by somatic cells such as fibroblasts and keratinocytes were inhibited in KO group (IL-1α: t_ (12h) (12h) = 5.049, P = 0.000; Cxcl-1: t_ (12h) = 3.187, P = 0.0004; t_ ) = 4.858, P = 0.000; Cxcl-2: t_ (24h) = 7.957, P = 0.000). Conclusion PLCε affects the inflammatory response by regulating somatic cytokine expression in Schistosoma japonicum cercarial dermatitis.