CP/CPPS模型大鼠Th1、Th17细胞L5~S2脊髓浸润及其中枢敏化机制研究

Infiltration of Th1 and Th17 cells into the L5-S2 spinal cord in CP/CPPS rats and its central sensitization mechanism

目的:探讨慢性前列腺炎/慢性骨盆疼痛综合征(CP/CPPS)疼痛的中枢敏化机制。方法:3~4周龄、体重250~350 g成年雄性SPF级SD大鼠40只随机均分为对照组和CP/CPPS模型组,构建模型成功后运用流式细胞术、实时荧光定量PCR(RT-qPCR)、免疫荧光检测等方法分析CD4^(+)T细胞亚群L5~S2脊髓的浸润情况,检测L5~S2脊髓GFAP、CR3的表达水平。结果:与对照组相比,CP/CPPS模型组L5~S2脊髓组织中单个核细胞中CD4^(+)T细胞表达量显著升高(P<0.01);RT-qPCR显示Th1细胞分泌的γ干扰素(IFN-γ)、肿瘤坏死因子α(TNF-α),Th17细胞分泌的白介素17(IL-17)和视黄酸相关孤儿受体γt(RORγt),细胞因子IL-6、IL-1β,趋化因子CCL2、CCL20、CXCL10等mRNA表达水平显著升高(P<0.01);免疫荧光染色显示Th1、Th17细胞的分子标志物IFN-γ、IL-17,星形胶质细胞和小胶质细胞的分子标志物胶质纤维酸性蛋白(GFAP)、补体受体3(CR3)荧光表达显著增强。结论:CP/CPPS模型大鼠L5~S2脊髓神经元中存在CD4^(+)T细胞浸润,具体亚群为Th1、Th17细胞,其分泌的炎症因子可能损伤神经元细胞并影响神经传导,促进中枢敏化并激活星形胶质细胞及小胶质细胞,导致疼痛的发生和发展。

Objective:To explore the central sensitization mechanism of pain in chronic prostatitis/chronic pelvic pain syndrome(CP/CPPS).Methods:We randomly divided 40 adult male SPF SD rats,aged 3-4 weeks and weighing 250-350 g,into a normal control and a CP/CPPS model group.After modeling,we analyzed the state of infiltration of CD4^(+)T cells into the L5-S2 spinal cord and detected the expression levels of GFAP and CR3 in the spinal cord tissue using flow cytometry,real-time fluorescent quantitative PCR(RT-qPCR)and immunofluorescence staining.Results:Compared with the normal controls,the CP/CPPS model rats showed dramatically increased expression of CD4^(+)T cells in the mononuclear cells of the L5-S2 spinal cord tissue(P<0.01),mRNA expressions of interferon-γ(IFN-γ)and tumor necrosis factorα(TNF-α)secreted from the Th1 cells,interleukin(IL)-17 and retinoic acid-associated orphan receptor(ROR)γt secreted from the Th17 cells,cytokines IL-6 and IL-1β,and chemokines CCL2,CCL20 and CXCL10(P<0.01),and expressions of the molecular markers of Th1 and Th17 cells IFN-γ and IL-17 and those of astrocytes and microglias GFAP and CR3.Conclusion:CD4^(+)T cells,specifically Th1 and Th17 cells,infiltrate L5-S2 spinal cord neurons in CP/CPPS model rats.The inflammatory factors secreted from these cells may damage the neuronal cells,affect nervous conduction,promote central sensitization and activate astrocytes and microglias,leading to the development and progression of pain.